The sympathetic nervous system plays a cardinal role in regulating cardiac function through releasing the neurotransmitter norepinephrine (NE). In comparison with central nervous system, the molecular mechanism of NE ...The sympathetic nervous system plays a cardinal role in regulating cardiac function through releasing the neurotransmitter norepinephrine (NE). In comparison with central nervous system, the molecular mechanism of NE uptake in myocardium is not clear. In present study, we proved that in rat the CNS type of NE transporter (NET) was also expressed in middle cervical-stellate ganglion complex (MC-SG complex) which is considered to control the activity of heart, but not expressed in myocardium. The results also showed that NET expression level in right ganglion was significantly higher than in the left, rendering the greater capacity of NE uptake in right ventricle, a fact which may contribute to the maintenance of right ventricular function under pathologic state.展开更多
The accumulation of excessive reactive oxygen species can exacerbate any injury of retinal tissue because free radicals can trigger lipid peroxidation,protein damage and DNA fragmentation.Increased oxidative stress is...The accumulation of excessive reactive oxygen species can exacerbate any injury of retinal tissue because free radicals can trigger lipid peroxidation,protein damage and DNA fragmentation.Increased oxidative stress is associated with the common pathological process of many eye diseases,such as glaucoma,diabetic retinopathy and ischemic optic neuropathy.Many studies have demonstrated that Lycium barbarum polysaccharides(LBP)protects against oxidative injury in numerous cells and tissues.For the model of hypoxia we used cultured retinal ganglion cells and induced hypoxia by incubating with 200μM cobalt chloride(CoCl2)for 24 hours.To investigate the protective effect of LBP and its mechanism of action against oxidative stress injury,the retinal tissue was pretreated with 0.5 mg/mL LBP for 24 hours.The results of flow cytometric analysis showed LBP could effectively reduce the CoCl2-induced retinal ganglion cell apoptosis,inhibited the generation of reactive oxygen species and the reduction of mitochondrial membrane potential.These findings suggested that LBP could protect retinal ganglion cells from CoCl2-induced apoptosis by reducing mitochondrial membrane potential and reactive oxygen species.展开更多
Human umbilical cord-derived mesenchymal stem cells （hUCMSCs） represent a promising young-state stem cell source for cell-based therapy. hUCMSC transplantation into the transected sciatic nerve promotes axonal regen...Human umbilical cord-derived mesenchymal stem cells （hUCMSCs） represent a promising young-state stem cell source for cell-based therapy. hUCMSC transplantation into the transected sciatic nerve promotes axonal regeneration and functional recovery. To further clarify the para-crine effects of hUCMSCs on nerve regeneration, we performed human cytokine antibody array analysis, which revealed that hUCMSCs express 14 important neurotrophic factors. Enzyme-linked immunosorbent assay and immunohistochemistry showed that brain-derived neurotrophic factor, glial-derived neurotrophic factor, hepatocyte growth factor, neurotrophin-3, basic fibroblast growth factor, type I collagen, fibronectin and laminin were highly expressed. Treatment with hUCMSC-conditioned medium enhanced Schwann cell viability and proliferation, increased nerve growth factor and brain-derived neurotrophic factor expression in Schwann cells, and enhanced neurite growth from dorsal root ganglion explants. These ifndings suggest that paracrine action may be a key mechanism underlying the effects of hUCMSCs in peripheral nerve repair.展开更多
To clarify the modulation of dragon's blood on the tetrodotoxin-resistant (TTX-R) sodium currents in dorsal root ganglion (DRG) neurons and explore its corresponding material basis for the efficacy, using whole-ce...To clarify the modulation of dragon's blood on the tetrodotoxin-resistant (TTX-R) sodium currents in dorsal root ganglion (DRG) neurons and explore its corresponding material basis for the efficacy, using whole-cell patch clamp technique, the effects of dragon's blood and the combined effects of three components (cochinchinenin A, cochinchinenin B, and loureirin B) extracted from dragon's blood on the TTX-R sodium currents in acute-isolated DRG neurons of rats were observed. According to the operational definition of material basis for the efficacy of TCM established, the material basis of the modulation on the TTX-R sodium currents in DRG neurons of dragon's blood was judged from the experimental results. The drug interaction equation of Greco et al. was used to assess the interaction of the three components extracted from dragon's blood. This investigation demonstrated that dragon's blood suppressed the peak TTX-R sodium currents in a dose-dependent way and affected the activations of TTX-R sodium currents. The effects of the combination of cochinchinenin A, cochinchinenin B, and loureirin B were in good agreement with those of dragon's blood. Although the three components used alone could modulate TTX-R sodium currents, the concentrations of the three components used alone were respectively higher than those used in combination when the inhibition rates on the TTX-R sodium currents of them used alone and in combination were the same. The combined effects of the three components were synergistic. These results suggested that the interference with pain messages caused by the modulation of dragon's blood on TTX-R sodium currents in DRG neurons may explain some of the analgesic effect of dragon's blood and the corresponding material basis for the efficacy is the combination of cochinchinenin A, cochinchinenin B, and loureirin B.展开更多
Background Glaucoma can cause progressive damage to retinal ganglion cells. These cells can be classified as cells projecting to the superior colliculus and melanopsin-containing retinal ganglion cells, which project ...Background Glaucoma can cause progressive damage to retinal ganglion cells. These cells can be classified as cells projecting to the superior colliculus and melanopsin-containing retinal ganglion cells, which project to the suprachiasmatic nucleus. This study was to investigate the effects of chronic intraocular pressure elevation on melanopsin-containing retinal ganglion cells in rats. Methods Chronic intraocular pressure elevation was induced in one eye of adult Wistar rats by cauterization of three episcleral veins. Intraocular pressure was measured at different intervals with a rebound tonometer. Superior collicular retinal ganglion cells were retrogradely labeled from the superior colliculus with Fluorogold. Melanopsin-containing retinal ganglion cells were visualized by free-floating immunohistochemistry on whole-mount retinas. The number of labeled superior collicular and melanopsin-containing retinal ganglion cells were counted in the sample areas on flat-mounted retinas. Results Compared with contralateral control eyes, the numbers of both superior collicular and melanopsin-containing retinal ganglion cells were significantly reduced after 12 weeks of experimental intraocular pressure elevation （（2317.41±29.96）/mm^2 vs （1815.82±24.25）/mm^2; （26.20±2.10）/mm^2 vs （20.62±1.52）/mm^2, respectively）. The extent of cell loss of the two types of retinal ganglion cells was similar. However, no morphologic changes were found in melanopsin-containing retinal ganglion cells. Conclusion Both melanopsin-containing and superior collicular retinal ganglion cells were damaged by chronic ocular hypertension, indicating that glaucomatous neural degeneration involves the non-image-forming visual pathway.展开更多
Percutaneous microballoon compression of the trigeminal ganglion is a brand new operative technique for the treatment of trigeminal neuralgia. However, it is unclear how the procedure mediates pain relief, and there a...Percutaneous microballoon compression of the trigeminal ganglion is a brand new operative technique for the treatment of trigeminal neuralgia. However, it is unclear how the procedure mediates pain relief, and there are no standardized criteria, such as compression pressure, com- pression time or balloon shape, for the procedure. In this study, percutaneous microballoon compression was performed on the rabbit trigeminal ganglion at a mean inflation pressure of 1,005 ＋ 150 mmHg for 2 or 5 minutes. At 1, 7 and 14 days after percutaneous microballoon compression, the large-diameter myelinated nerves displayed axonal swelling, rupture and demy- elination under the electron microscope. Fragmentation of myelin and formation of digestion chambers were more evident after 5 minutes of compression. Image analyzer results showed that the diameter of trigeminal ganglion cells remained unaltered after compression. These experi- mental findings indicate that a 2-minute period of compression can suppress pain transduction. Immunohistochemical staining revealed that vascular endothelial growth factor expression in the ganglion cells and axons was significantly increased 7 days after trigeminal ganglion compression, however, the changes were similar after 2-minute compression and 5-minute compression. The upregulated expression of vascular endothelial growth factor in the ganglion cells after percu- taneous microballoon compression can promote the repair of the injured nerve. These findings suggest that long-term compression is ideal for patients with recurrent trigeminal neuralgia.展开更多
The development, cytological characters and ultrastructure of phloem ganglion in the nodal region of Phyllostachys edulis (Carr.) H. de Lehaie, a most economically important bamboo, were investigated and the possi...The development, cytological characters and ultrastructure of phloem ganglion in the nodal region of Phyllostachys edulis (Carr.) H. de Lehaie, a most economically important bamboo, were investigated and the possible physiological function of this special structure was proposed. The phloem ganglion derived directly from procambium is situated at the sites where the vascular bundle forks and is present in pairs. The phloem ganglion is spindle_like in appearance and usually consists of 4 to 6 layers. Two kinds of cells in the ganglion could be distinguished. In the middle, there are two layers of filiform cells with pointed ends so that there are no normal sieve plates. Nevertheless, there are many pits on the lateral wall of the filiform cells. The other type of cells located at both ends of the spindle which possess an intermediate form between the filiform cell and the normal sieve tube. The walls of these cells towards the filiform cells are strongly convex forming a special sieve plate. Ultrastructure study showed that cells in the ganglion are connected by enriched plasmodesma. During early differentiation, the paramural body and the ingrowth of cell wall could be observed. It indicates that the cells of phloem ganglion have the character of transfer cells. The organelles in the mature cells are mainly plastids with abundant accumulation of proteins of crystalline structure. The above_mentioned results suggest that the physiological function of the phloem ganglion is closely related with substance transport.展开更多
Using whole-cell patch clamp technique on the membrane of freshly isolated dorsal root ganglion (DRG) neurons, the effects of dragons blood resin and its important component loureirin B on tetrodotoxin-sensitive (TTX-...Using whole-cell patch clamp technique on the membrane of freshly isolated dorsal root ganglion (DRG) neurons, the effects of dragons blood resin and its important component loureirin B on tetrodotoxin-sensitive (TTX-S) voltage-gated sodium currents were observed. The results show that both blood resin and loureirin B could suppress TTX-S voltage-gated sodium currents in a dose-dependent way. The peak current amplitudes and the steady-state activation and inactivation curves are also made to shift by 0.05% blood resin and 0.2 mmol/L loureirin B. These results demonstrate that the effects of blood resin on TTX-S sodium current may contrib-ute to loureirin B in blood resin. Perhaps the analgesic effect of blood resin is caused partly by loureirin B directly interfering with the nociceptive transmission of primary sensory neurons.展开更多
Patients treated with platinum-based chemotherapy frequently experience neurotoxic symptoms, which may lead to premature discontinuation of therapy. Despitediscontinuation of platinum drugs, these symptoms can persist...Patients treated with platinum-based chemotherapy frequently experience neurotoxic symptoms, which may lead to premature discontinuation of therapy. Despitediscontinuation of platinum drugs, these symptoms can persist over a long period of time. Cisplatin and oxaliplatin, among all platinum drugs, have significant neurotoxic potential. A distal dose-dependent symmetrical sensory neuropathy is the most common presentation of platinum neurotoxicity. DNA damage-induced apoptosis of dorsal root ganglion(DRG) neurons seems to be the principal cause of neurological symptoms. However, DRG injury alone cannot explain some unique symptoms such as cold-aggravated burning pain affecting distal extremities that is observed with oxaliplatin administration. In this article, we briefly reviewed potential mechanisms for the development of platinum drugs-associated neurological manifestations.展开更多
Lycium barbarum, a traditional Chinese anti-aging herb, has been shown to protect retinal ganglion cells （RGCs） in a rat chronic ocular hypertension （COH） model. Here, we investigated the expression of endothelin...Lycium barbarum, a traditional Chinese anti-aging herb, has been shown to protect retinal ganglion cells （RGCs） in a rat chronic ocular hypertension （COH） model. Here, we investigated the expression of endothelin-1 （ET-1）, a strong vasoconstrictor, and its receptors, ETA and ETB, in the COH model and assessed the effects of Lycium barbarum on the ET-1 axis. Elevated intraocular pressure （IOP） was induced in the right eye of SD rats using argon laser photocoagulation. （1） The expression of ET-1, ETA and ETB in normal and COH retinas was studied. （2） Some COH rats were fed daily with Lycium barbarum Polysaccharides （LBP） using 1 mg/kg or phosphate-buffered saline （PBS） for 3 weeks （started 1 week before photocoagulation）. The effects of LBP on the expression of ET-1 and its receptors, ETA and ETB, in COH retina were evaluated. A semi-quantitative analysis of staining intensity was used to evaluate the expression levels of ET-1, ETA and ETB in retinal vasculature. We found that （1） Under COH condition, the immunoreactivity of ET-1 was increased in retina associated with an increase of ETB receptor immunoreactivity and a decrease of ETA receptor immunoreactivity. （2） After feeding COH rats with LBP, the expression of ET-1 was decreased with an increase of ETA expression and a decrease of ETB expression in the retina, especially in RGCs. （3） By comparing the staining intensity in the vasculature of COH retina in LBP-fed group with PBS-fed group, there was a decrease in the expression of ET-1 and ETA and an increase in ETB. In summary, ET-1 expression was up-regulated in the retina in COH model. LBP could decrease the expression of ET-1 and modulate the expression of its receptors, ETA and ETB, under the condition of COH. The neuroprotective effect of LBP on RGCs might be related to its ability to regulate the ET-1-mediated biological effects on RGCs and retinal vasculature.展开更多
Puerarin is a major active ingredient of the traditional Chinese plant medicine,Radix Puerariae,and commonly used in the treatment of myocardial and cerebral ischemia.However,the effects of puerarin on neuropathic pai...Puerarin is a major active ingredient of the traditional Chinese plant medicine,Radix Puerariae,and commonly used in the treatment of myocardial and cerebral ischemia.However,the effects of puerarin on neuropathic pain are still unclear.In this study,a neuropathic pain animal model was created by partial sciatic nerve ligation.Puerarin（30 or 60 mg/kg） was intraperitoneally injected once a day for 7 days.Mechanical allodynia and thermal hyperalgesia were examined at 1 day after model establishment.Mechanical threshold and paw withdrawal latency markedly increased in a dose-dependent manner in puerarin-treated rats,especially at 7 days after model establishment.At 7 days after model establishment,quantitative real-time reverse transcriptase-polymerase chain reaction results showed that puerarin administration reversed m RNA expression of transient receptor potential vanilloid 1（Trpv1） and transient receptor potential ankyrin 1（Trpa1） in a dose-dependent manner in dorsal root ganglion neurons after peripheral nerve injury.These results suggest that puerarin dose-dependently ameliorates neuropathic pain by suppressing Trpv1 and Trpa1 up-regulation in dorsal root ganglion of neuropathic pain rats.展开更多
Voltage-gated sodium channels （Navs） play an important role in human pain sensation. However, the expression and role of Nav subtypes in native human sensory neurons are unclear. To address this issue, we obtained h...Voltage-gated sodium channels （Navs） play an important role in human pain sensation. However, the expression and role of Nav subtypes in native human sensory neurons are unclear. To address this issue, we obtained human dorsal root ganglion （hDRG） tissues from healthy donors. PCR analysis of seven DRG-expressed Nav subtypes revealed that the hDRG has higher expression of Navl.7 （,-~ 50% of total Nav expression） and lower expres- sion of Navl.8 （~ 12%）, whereas the mouse DRG has higher expression of Nav 1.8 （- 45%） and lower expression of Navl.7 （- 18%）. To mimic Nav regulation in chronic pain, we treated hDRG neurons in primary cultures with paclitaxel （0.1-1 μmol/L） for 24 h. Paclitaxel increased the Navl.7 but not Navl.8 expression and also increased the transient Na＋ currents and action potential firing frequency in small-diameter （〈50 ~tm） hDRG neurons. Thus, the hDRG provides a translational model in which to study ＂human pain in a dish＂ and test new pain therapeutics.展开更多
AIM To investigate the efficacy and safety of stellate ganglion block for the treatment of patients with chronic ulcerative colitis.METHODS A total of 120 randomly selected patients with chronic ulcerative colitis tre...AIM To investigate the efficacy and safety of stellate ganglion block for the treatment of patients with chronic ulcerative colitis.METHODS A total of 120 randomly selected patients with chronic ulcerative colitis treated in Cangzhou Central Hospital from January 2014 to January 2016 were included in this study. These patients were divided into two groups: control group(n = 30), patients received oral sulfasalazine treatment; experimental group(n = 90), patients received stellate ganglion block treatment. Clinical symptoms and disease activity in these two groups were compared before and after treatment using endoscopy. Blood was collected from patients on day 0, 10, 20 and 30 after treatment. Enzyme-linked immunosorbent assay was performed to determine interleukin-8(IL-8) level. The changes in IL-8 level post-treatment in the two groups were compared using repeated measures analysis of variance.RESULTS After treatment, clinical symptoms and disease activity were shown to be alleviated by endoscopy in both the control and experimental groups. However, patients in the control group did not have obvious abdominal pain relief. In addition, the degree of pain relief in the experimental group was statistically better than that in the control group(P < 0.05). Ten days after treatment, IL-8 level was found to be significantly lower in the experimental group than in the control group, and the difference was statistically significant(P < 0.05). In addition, adverse events were significantly higher in the control group than in the experimental group, and the difference was statistically significant(χ~2 = 33.215, P = 0.000).CONCLUSION The application of stellate ganglion block effectively improves treatment efficacy in chronic ulcerative colitis, relieves clinical symptoms in patients, and reduces the level of inflammatory factors. Furthermore, this approach also had a positive impact on the disease to a certain extent.展开更多
Orthodontic pain is an inflammatory pain that is initiated by orthodontic force-induced vascular occlusion followed by a cascade of inflammatory responses, including vascular changes, the recruitment of inflammatory a...Orthodontic pain is an inflammatory pain that is initiated by orthodontic force-induced vascular occlusion followed by a cascade of inflammatory responses, including vascular changes, the recruitment of inflammatory and immune cells, and the release of neurogenic and pro-inflammatory mediators. Ultimately, endogenous analgesic mechanisms check the inflammatory response and the sensation of pain subsides. The orthodontic pain signal, once received by periodontal sensory endings, reaches the sensory cortex for pain perception through three-order neurons： the trigeminal neuron at the trigeminal ganglia, the trigeminal nucleus caudalis at the medulla oblongata and the ventroposterior nucleus at the thalamus. Many brain areas participate in the emotion, cognition and memory of orthodontic pain, including the insular cortex, amygdala, hippocampus, locus coeruleus and hypothalamus. A built-in analgesic neural pathway--periaqueductal grey and dorsal raphe---has an important role in alleviating orthodontic pain. Currently, several treatment modalities have been applied for the relief of orthodontic pain, including pharmacological, mechanical and behavioural approaches and low-level laser therapy. The effectiveness of nonsteroidal anti- inflammatory drugs for pain relief has been validated, but its effects on tooth movement are controversial. However, more studies are needed to verify the effectiveness of other modalities. Furthermore, gene therapy is a novel, viable and promising modality for alleviatin~ orthodontic oain in the future.展开更多
Objective:To investigate protective effects of hirudin on oxidative stress and apoptosis of spinal dorsal root ganglion cells in high-glucose rats at the cellular and molecular level.Methods:Dorsal root ganglion neuro...Objective:To investigate protective effects of hirudin on oxidative stress and apoptosis of spinal dorsal root ganglion cells in high-glucose rats at the cellular and molecular level.Methods:Dorsal root ganglion neurons(DRGn)were harvested from embryonic day in 15 SD rats,purified and identificated after primary culture.They were divided into the normal control group,high-glucose(HG)group,positive control(alpha-lipoic acid,ALA)group,low-dose hirudin group(H1),medium-dose hirudin group(H2)and high-dose hirudin group(H3).The control group was cultured by neuron specific culture medium,while the HG group was cultured by neuron specific culture medium and 20 mmol/L glucose(HG medium).The hirudin groups were cultured by HG medium+0.25 IU/mL hirudin(H1),HG medium+0.5 IU/mL hirudin(H2)and HG medium+1 IU/mL hirudin(H3).The ALA group was cultured by HG medium +100μmol/L ALA.3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylt etrazolium bromide(MTT)assay was used to explore the optimum concentration and intervention time.Flow cytometry assay was used to detect the level of reactive oxygen series(ROS).Western blot and quantificational realtime polymerase chain reaction(qRT-PCR)were used to detect the expression of protein and mRNA of nuclear factor erythroid 2-related factor 2(Nrf-2),hemeoxygence-1(HO-1),nuclear factor-κB(NF-κB)and Caspase-3.TUNEL assay was used to test the apoptosis rate of different groups.Results:After 24 h of culture,the cell activity of hirudin and ALA groups were higher than that of HG group,and there was a statistical difference between the H1 group and HG group(P<0.05).In hirudin groups,the apoptosis rate of cells,the expression of activated Caspase-3 protein and Caspase-3 mRNA were lower than those of HG group(P<0.01),higher than those of ALA group(P<0.01 or P<0.05).The ROS level of hirudin groups was higher than that of ALA group(P<0.01),lower than that of HG group(P<0.01 or P<0.05).The expression of NF-κB(P65)protein in H3 group were lower than those of HG group(P<0.05).The expression of Nrf-2 protein展开更多
Objective： TO examine the effects of the combination of quercetin （Q）, cinnamaldehyde （C） and hirudin （H）, a Chinese medicine formula on high glucose （HG）-induced apoptosis of cultured dorsal root ganglion （...Objective： TO examine the effects of the combination of quercetin （Q）, cinnamaldehyde （C） and hirudin （H）, a Chinese medicine formula on high glucose （HG）-induced apoptosis of cultured dorsal root ganglion （DRG） neurons. Methods： DRG neurons exposed to HG （45 mmol/L） for 24 h were employed as an in vitro model of diabetic neuropathy. Cell viability, reactive oxygen species （ROS） level and apoptosis were determined. The expression of nuclear factor of Kappa B （NF- κB）, inhibitory kappa B α （IκBα ）, phosphorylated IκBα and Nf-E2 related factor 2 （Nrf2） were examined using reverse transcription-polymerase chain reaction （RT-PCR） and Western blot assay. The expression of hemeoxygenase-1 （HO-1）, interleukin-6 （IL-6）, tumor necrosis factor （TNF-α） and caspase-3 were also examined by RT-PCR and Western blot assay. Results： HG treatment markedly increased DRG neuron apoptosis via increasing intracellular ROS level and activating the NF- κB signaling pathway （P〈0.05）. Co-treatment with Q, C, H and their combination decreased HG-induced caspase-3 activation and apoptosis （P〈0.05 or P〈0.01）. The expressions of NF- κB, IL-6 and TNF-α were down- regulated, and Nrf2/HO-1 expression was up-regulated （P〈0.05 or P〈0.01）. QCH has better effect in scavenging ROS, activating Nrf-2/HO-1, and down-regulating the NF- κB pathway than other treatment group. Conclusions： DRG neurons＇ apoptosis was increased in diabetic conditions, which was reduced by QCH formula treatment. The possible reason could be activating Nrf-2/HO-1 pathway, scavenging ROS, and inhibition of NF- κB activation. The effect of QCH combination was better than each monomer or the combination of the two monomers.展开更多
Astragaloside Ⅳ is the main active compound of Astragalus membranaceus. Astragaloside Ⅳ has strong anti-oxidative activities and protective effects against progression of peripheral neuropathy. In this study, we det...Astragaloside Ⅳ is the main active compound of Astragalus membranaceus. Astragaloside Ⅳ has strong anti-oxidative activities and protective effects against progression of peripheral neuropathy. In this study, we determined whether astragaloside Ⅳ protects retinal ganglion cells（RGC） from oxidative stress injury using the rat RGC-5 cell line. Hydrogen peroxide（H_2O_2） was used to induce oxidative stress injury, with the protective effect of astragaloside Ⅳ examined. Cell Counting Kit-8 and 4′,6-diamidino-2-phenylindole staining showed that astragaloside Ⅳ increased cell survival rate and decreased apoptotic cell number. Flow cytometry showed that astragaloside Ⅳ decreased H_2O_2-induced reactive oxygen species levels. While laser confocal microscopy showed that astragaloside Ⅳ inhibited the H_2O_2-induced decrease of mitochondrial membrane potential. Western blot assay showed that astragaloside Ⅳ reduced cytochrome c release induced by H_2O_2, inhibited Bax and caspase-3 expression, and increased Bcl-2 expression. Altogether, these results indicate that astragaloside Ⅳ has potential protective effects against H_2O_2-induced oxidative stress in retinal ganglion cells.展开更多
Objective： To examine the mechanism underlying the beneficial role of cinnamaldehyde on oxidative damage and apoptosis in high glucose （HG）-induced dorsal root ganglion （DRG） neurons in vitro. Methods： HG-treate...Objective： To examine the mechanism underlying the beneficial role of cinnamaldehyde on oxidative damage and apoptosis in high glucose （HG）-induced dorsal root ganglion （DRG） neurons in vitro. Methods： HG-treated DRG neurons were developed as an in vitro model of diabetic neuropathy. The neurons were randomly divided into five groups： the control group, the HG group and the HG groups treated with 25, 50 and 100 nmol/L cinnamaldehyde, respectively. Cell viability was examined by 3-（4,5-dimethylthiazol-2-yl）-2,5- diphenyltetrazolium bromide （MTT） assay and apoptosis rate was evaluated by the in situ TdT-mediated dUTP nick end labeling （TUNEL） assay. The intracellular level of reactive oxygen species （ROS） was measured with flow cytometry. Expression of nuclear factor-kappa B （NF-κ B）, inhibitor of κ B （I κ B）, phosphorylated I κ B （p-IκB）, tumor necrosis factor （TNF）-α, interleukin-6 （IL-6） and caspase-3 were determined by western blotting and real-time quantitative reverse transcription polymerase chain reaction （RT-PCR）. Expression of nuclear factor erythroid 2-related factor 2 （Nrf2） and hemeoxygenase-1 （HO-1） were also measured by western blotting. Results： Cinnamaldehyde reduced HG-induced loss of viability, apoptosis and intracellular generation of ROS in the DRG neurons via inhibiting NF- K B activity. The western blot assay results showed that the HG-induced elevated expressions of NF- κB, I κ B and p-I κ B were remarkably reduced by cinnamaldehyde treatment in a dose-dependent manner （P〈0.01）. The HG-induced over-expression of NF-κ B p65 mRNA was remarkably attenuated after cinnamaldehyde treatment in a dose-dependent manner （P〈0.01）. However, the expressions of Nrf2 and HO-1 were not upregulated. Treatment with cinnamaldehyde not only attenuated caspase-3 activation and the caspase cleavage cascade in DRG neurons, but also lowered the elevated IL-6, TNF-α, cyclo-oxygenase and inducible nitric oxide synthase levels, indi展开更多
文摘The sympathetic nervous system plays a cardinal role in regulating cardiac function through releasing the neurotransmitter norepinephrine (NE). In comparison with central nervous system, the molecular mechanism of NE uptake in myocardium is not clear. In present study, we proved that in rat the CNS type of NE transporter (NET) was also expressed in middle cervical-stellate ganglion complex (MC-SG complex) which is considered to control the activity of heart, but not expressed in myocardium. The results also showed that NET expression level in right ganglion was significantly higher than in the left, rendering the greater capacity of NE uptake in right ventricle, a fact which may contribute to the maintenance of right ventricular function under pathologic state.
基金supported by grants from Project of Administration of Traditional Chinese Medicine of Guangdong Province of China,No.20161071(to LL)Medical Scientific Research Foundation of Guangdong Province of China,No.A2019098(to LL)
文摘The accumulation of excessive reactive oxygen species can exacerbate any injury of retinal tissue because free radicals can trigger lipid peroxidation,protein damage and DNA fragmentation.Increased oxidative stress is associated with the common pathological process of many eye diseases,such as glaucoma,diabetic retinopathy and ischemic optic neuropathy.Many studies have demonstrated that Lycium barbarum polysaccharides(LBP)protects against oxidative injury in numerous cells and tissues.For the model of hypoxia we used cultured retinal ganglion cells and induced hypoxia by incubating with 200μM cobalt chloride(CoCl2)for 24 hours.To investigate the protective effect of LBP and its mechanism of action against oxidative stress injury,the retinal tissue was pretreated with 0.5 mg/mL LBP for 24 hours.The results of flow cytometric analysis showed LBP could effectively reduce the CoCl2-induced retinal ganglion cell apoptosis,inhibited the generation of reactive oxygen species and the reduction of mitochondrial membrane potential.These findings suggested that LBP could protect retinal ganglion cells from CoCl2-induced apoptosis by reducing mitochondrial membrane potential and reactive oxygen species.
基金supported by the National Natural Science Foundation of China,No.31100696,31170946a grant from the National High Technology Research and Development Program of China(863 Program),No.2012AA020502+1 种基金a grant from the National Program on Key Basic Research Project of China(973 Program),No.2014CB542201a grant from Beijing Metropolis Beijing Nova Program,No.2011115
文摘Human umbilical cord-derived mesenchymal stem cells （hUCMSCs） represent a promising young-state stem cell source for cell-based therapy. hUCMSC transplantation into the transected sciatic nerve promotes axonal regeneration and functional recovery. To further clarify the para-crine effects of hUCMSCs on nerve regeneration, we performed human cytokine antibody array analysis, which revealed that hUCMSCs express 14 important neurotrophic factors. Enzyme-linked immunosorbent assay and immunohistochemistry showed that brain-derived neurotrophic factor, glial-derived neurotrophic factor, hepatocyte growth factor, neurotrophin-3, basic fibroblast growth factor, type I collagen, fibronectin and laminin were highly expressed. Treatment with hUCMSC-conditioned medium enhanced Schwann cell viability and proliferation, increased nerve growth factor and brain-derived neurotrophic factor expression in Schwann cells, and enhanced neurite growth from dorsal root ganglion explants. These ifndings suggest that paracrine action may be a key mechanism underlying the effects of hUCMSCs in peripheral nerve repair.
文摘To clarify the modulation of dragon's blood on the tetrodotoxin-resistant (TTX-R) sodium currents in dorsal root ganglion (DRG) neurons and explore its corresponding material basis for the efficacy, using whole-cell patch clamp technique, the effects of dragon's blood and the combined effects of three components (cochinchinenin A, cochinchinenin B, and loureirin B) extracted from dragon's blood on the TTX-R sodium currents in acute-isolated DRG neurons of rats were observed. According to the operational definition of material basis for the efficacy of TCM established, the material basis of the modulation on the TTX-R sodium currents in DRG neurons of dragon's blood was judged from the experimental results. The drug interaction equation of Greco et al. was used to assess the interaction of the three components extracted from dragon's blood. This investigation demonstrated that dragon's blood suppressed the peak TTX-R sodium currents in a dose-dependent way and affected the activations of TTX-R sodium currents. The effects of the combination of cochinchinenin A, cochinchinenin B, and loureirin B were in good agreement with those of dragon's blood. Although the three components used alone could modulate TTX-R sodium currents, the concentrations of the three components used alone were respectively higher than those used in combination when the inhibition rates on the TTX-R sodium currents of them used alone and in combination were the same. The combined effects of the three components were synergistic. These results suggested that the interference with pain messages caused by the modulation of dragon's blood on TTX-R sodium currents in DRG neurons may explain some of the analgesic effect of dragon's blood and the corresponding material basis for the efficacy is the combination of cochinchinenin A, cochinchinenin B, and loureirin B.
基金This study was supported by a grant from the National Natural Science Foundation of China （No. 30571991）.
文摘Background Glaucoma can cause progressive damage to retinal ganglion cells. These cells can be classified as cells projecting to the superior colliculus and melanopsin-containing retinal ganglion cells, which project to the suprachiasmatic nucleus. This study was to investigate the effects of chronic intraocular pressure elevation on melanopsin-containing retinal ganglion cells in rats. Methods Chronic intraocular pressure elevation was induced in one eye of adult Wistar rats by cauterization of three episcleral veins. Intraocular pressure was measured at different intervals with a rebound tonometer. Superior collicular retinal ganglion cells were retrogradely labeled from the superior colliculus with Fluorogold. Melanopsin-containing retinal ganglion cells were visualized by free-floating immunohistochemistry on whole-mount retinas. The number of labeled superior collicular and melanopsin-containing retinal ganglion cells were counted in the sample areas on flat-mounted retinas. Results Compared with contralateral control eyes, the numbers of both superior collicular and melanopsin-containing retinal ganglion cells were significantly reduced after 12 weeks of experimental intraocular pressure elevation （（2317.41±29.96）/mm^2 vs （1815.82±24.25）/mm^2; （26.20±2.10）/mm^2 vs （20.62±1.52）/mm^2, respectively）. The extent of cell loss of the two types of retinal ganglion cells was similar. However, no morphologic changes were found in melanopsin-containing retinal ganglion cells. Conclusion Both melanopsin-containing and superior collicular retinal ganglion cells were damaged by chronic ocular hypertension, indicating that glaucomatous neural degeneration involves the non-image-forming visual pathway.
基金supported by a grant from Shengjing Hospital,China Medical University,China,No.201010252
文摘Percutaneous microballoon compression of the trigeminal ganglion is a brand new operative technique for the treatment of trigeminal neuralgia. However, it is unclear how the procedure mediates pain relief, and there are no standardized criteria, such as compression pressure, com- pression time or balloon shape, for the procedure. In this study, percutaneous microballoon compression was performed on the rabbit trigeminal ganglion at a mean inflation pressure of 1,005 ＋ 150 mmHg for 2 or 5 minutes. At 1, 7 and 14 days after percutaneous microballoon compression, the large-diameter myelinated nerves displayed axonal swelling, rupture and demy- elination under the electron microscope. Fragmentation of myelin and formation of digestion chambers were more evident after 5 minutes of compression. Image analyzer results showed that the diameter of trigeminal ganglion cells remained unaltered after compression. These experi- mental findings indicate that a 2-minute period of compression can suppress pain transduction. Immunohistochemical staining revealed that vascular endothelial growth factor expression in the ganglion cells and axons was significantly increased 7 days after trigeminal ganglion compression, however, the changes were similar after 2-minute compression and 5-minute compression. The upregulated expression of vascular endothelial growth factor in the ganglion cells after percu- taneous microballoon compression can promote the repair of the injured nerve. These findings suggest that long-term compression is ideal for patients with recurrent trigeminal neuralgia.
基金InternationalFoundationforScience Sweden +1 种基金D/ 2 1 69_1 FoundationoftheEducationMinistryofChina .
文摘The development, cytological characters and ultrastructure of phloem ganglion in the nodal region of Phyllostachys edulis (Carr.) H. de Lehaie, a most economically important bamboo, were investigated and the possible physiological function of this special structure was proposed. The phloem ganglion derived directly from procambium is situated at the sites where the vascular bundle forks and is present in pairs. The phloem ganglion is spindle_like in appearance and usually consists of 4 to 6 layers. Two kinds of cells in the ganglion could be distinguished. In the middle, there are two layers of filiform cells with pointed ends so that there are no normal sieve plates. Nevertheless, there are many pits on the lateral wall of the filiform cells. The other type of cells located at both ends of the spindle which possess an intermediate form between the filiform cell and the normal sieve tube. The walls of these cells towards the filiform cells are strongly convex forming a special sieve plate. Ultrastructure study showed that cells in the ganglion are connected by enriched plasmodesma. During early differentiation, the paramural body and the ingrowth of cell wall could be observed. It indicates that the cells of phloem ganglion have the character of transfer cells. The organelles in the mature cells are mainly plastids with abundant accumulation of proteins of crystalline structure. The above_mentioned results suggest that the physiological function of the phloem ganglion is closely related with substance transport.
文摘Using whole-cell patch clamp technique on the membrane of freshly isolated dorsal root ganglion (DRG) neurons, the effects of dragons blood resin and its important component loureirin B on tetrodotoxin-sensitive (TTX-S) voltage-gated sodium currents were observed. The results show that both blood resin and loureirin B could suppress TTX-S voltage-gated sodium currents in a dose-dependent way. The peak current amplitudes and the steady-state activation and inactivation curves are also made to shift by 0.05% blood resin and 0.2 mmol/L loureirin B. These results demonstrate that the effects of blood resin on TTX-S sodium current may contrib-ute to loureirin B in blood resin. Perhaps the analgesic effect of blood resin is caused partly by loureirin B directly interfering with the nociceptive transmission of primary sensory neurons.
文摘Patients treated with platinum-based chemotherapy frequently experience neurotoxic symptoms, which may lead to premature discontinuation of therapy. Despitediscontinuation of platinum drugs, these symptoms can persist over a long period of time. Cisplatin and oxaliplatin, among all platinum drugs, have significant neurotoxic potential. A distal dose-dependent symmetrical sensory neuropathy is the most common presentation of platinum neurotoxicity. DNA damage-induced apoptosis of dorsal root ganglion(DRG) neurons seems to be the principal cause of neurological symptoms. However, DRG injury alone cannot explain some unique symptoms such as cold-aggravated burning pain affecting distal extremities that is observed with oxaliplatin administration. In this article, we briefly reviewed potential mechanisms for the development of platinum drugs-associated neurological manifestations.
基金supported by the Azalea (1972) Education fund to KFSo and RCCCFundamental Research Fund for The Centre Universities,No.21609101
文摘Lycium barbarum, a traditional Chinese anti-aging herb, has been shown to protect retinal ganglion cells （RGCs） in a rat chronic ocular hypertension （COH） model. Here, we investigated the expression of endothelin-1 （ET-1）, a strong vasoconstrictor, and its receptors, ETA and ETB, in the COH model and assessed the effects of Lycium barbarum on the ET-1 axis. Elevated intraocular pressure （IOP） was induced in the right eye of SD rats using argon laser photocoagulation. （1） The expression of ET-1, ETA and ETB in normal and COH retinas was studied. （2） Some COH rats were fed daily with Lycium barbarum Polysaccharides （LBP） using 1 mg/kg or phosphate-buffered saline （PBS） for 3 weeks （started 1 week before photocoagulation）. The effects of LBP on the expression of ET-1 and its receptors, ETA and ETB, in COH retina were evaluated. A semi-quantitative analysis of staining intensity was used to evaluate the expression levels of ET-1, ETA and ETB in retinal vasculature. We found that （1） Under COH condition, the immunoreactivity of ET-1 was increased in retina associated with an increase of ETB receptor immunoreactivity and a decrease of ETA receptor immunoreactivity. （2） After feeding COH rats with LBP, the expression of ET-1 was decreased with an increase of ETA expression and a decrease of ETB expression in the retina, especially in RGCs. （3） By comparing the staining intensity in the vasculature of COH retina in LBP-fed group with PBS-fed group, there was a decrease in the expression of ET-1 and ETA and an increase in ETB. In summary, ET-1 expression was up-regulated in the retina in COH model. LBP could decrease the expression of ET-1 and modulate the expression of its receptors, ETA and ETB, under the condition of COH. The neuroprotective effect of LBP on RGCs might be related to its ability to regulate the ET-1-mediated biological effects on RGCs and retinal vasculature.
基金supported by the National Natural Science Foundation of China,No.81671891
文摘Puerarin is a major active ingredient of the traditional Chinese plant medicine,Radix Puerariae,and commonly used in the treatment of myocardial and cerebral ischemia.However,the effects of puerarin on neuropathic pain are still unclear.In this study,a neuropathic pain animal model was created by partial sciatic nerve ligation.Puerarin（30 or 60 mg/kg） was intraperitoneally injected once a day for 7 days.Mechanical allodynia and thermal hyperalgesia were examined at 1 day after model establishment.Mechanical threshold and paw withdrawal latency markedly increased in a dose-dependent manner in puerarin-treated rats,especially at 7 days after model establishment.At 7 days after model establishment,quantitative real-time reverse transcriptase-polymerase chain reaction results showed that puerarin administration reversed m RNA expression of transient receptor potential vanilloid 1（Trpv1） and transient receptor potential ankyrin 1（Trpa1） in a dose-dependent manner in dorsal root ganglion neurons after peripheral nerve injury.These results suggest that puerarin dose-dependently ameliorates neuropathic pain by suppressing Trpv1 and Trpa1 up-regulation in dorsal root ganglion of neuropathic pain rats.
基金supported in part by NIH RO1Grants NS87988,DE17794,and DE22743 to R.R.J and NS89479 to S.Y.L and R.R.J
文摘Voltage-gated sodium channels （Navs） play an important role in human pain sensation. However, the expression and role of Nav subtypes in native human sensory neurons are unclear. To address this issue, we obtained human dorsal root ganglion （hDRG） tissues from healthy donors. PCR analysis of seven DRG-expressed Nav subtypes revealed that the hDRG has higher expression of Navl.7 （,-~ 50% of total Nav expression） and lower expres- sion of Navl.8 （~ 12%）, whereas the mouse DRG has higher expression of Nav 1.8 （- 45%） and lower expression of Navl.7 （- 18%）. To mimic Nav regulation in chronic pain, we treated hDRG neurons in primary cultures with paclitaxel （0.1-1 μmol/L） for 24 h. Paclitaxel increased the Navl.7 but not Navl.8 expression and also increased the transient Na＋ currents and action potential firing frequency in small-diameter （〈50 ~tm） hDRG neurons. Thus, the hDRG provides a translational model in which to study ＂human pain in a dish＂ and test new pain therapeutics.
文摘AIM To investigate the efficacy and safety of stellate ganglion block for the treatment of patients with chronic ulcerative colitis.METHODS A total of 120 randomly selected patients with chronic ulcerative colitis treated in Cangzhou Central Hospital from January 2014 to January 2016 were included in this study. These patients were divided into two groups: control group(n = 30), patients received oral sulfasalazine treatment; experimental group(n = 90), patients received stellate ganglion block treatment. Clinical symptoms and disease activity in these two groups were compared before and after treatment using endoscopy. Blood was collected from patients on day 0, 10, 20 and 30 after treatment. Enzyme-linked immunosorbent assay was performed to determine interleukin-8(IL-8) level. The changes in IL-8 level post-treatment in the two groups were compared using repeated measures analysis of variance.RESULTS After treatment, clinical symptoms and disease activity were shown to be alleviated by endoscopy in both the control and experimental groups. However, patients in the control group did not have obvious abdominal pain relief. In addition, the degree of pain relief in the experimental group was statistically better than that in the control group(P < 0.05). Ten days after treatment, IL-8 level was found to be significantly lower in the experimental group than in the control group, and the difference was statistically significant(P < 0.05). In addition, adverse events were significantly higher in the control group than in the experimental group, and the difference was statistically significant(χ~2 = 33.215, P = 0.000).CONCLUSION The application of stellate ganglion block effectively improves treatment efficacy in chronic ulcerative colitis, relieves clinical symptoms in patients, and reduces the level of inflammatory factors. Furthermore, this approach also had a positive impact on the disease to a certain extent.
基金supported by the National Natural Science Foundation of China (81571004, 81500884 and 81400549)the Orthodontic National Key Clinical Specialty Construction Program of China, West China Hospital of Stomatology, Sichuan University
文摘Orthodontic pain is an inflammatory pain that is initiated by orthodontic force-induced vascular occlusion followed by a cascade of inflammatory responses, including vascular changes, the recruitment of inflammatory and immune cells, and the release of neurogenic and pro-inflammatory mediators. Ultimately, endogenous analgesic mechanisms check the inflammatory response and the sensation of pain subsides. The orthodontic pain signal, once received by periodontal sensory endings, reaches the sensory cortex for pain perception through three-order neurons： the trigeminal neuron at the trigeminal ganglia, the trigeminal nucleus caudalis at the medulla oblongata and the ventroposterior nucleus at the thalamus. Many brain areas participate in the emotion, cognition and memory of orthodontic pain, including the insular cortex, amygdala, hippocampus, locus coeruleus and hypothalamus. A built-in analgesic neural pathway--periaqueductal grey and dorsal raphe---has an important role in alleviating orthodontic pain. Currently, several treatment modalities have been applied for the relief of orthodontic pain, including pharmacological, mechanical and behavioural approaches and low-level laser therapy. The effectiveness of nonsteroidal anti- inflammatory drugs for pain relief has been validated, but its effects on tooth movement are controversial. However, more studies are needed to verify the effectiveness of other modalities. Furthermore, gene therapy is a novel, viable and promising modality for alleviatin~ orthodontic oain in the future.
基金Supported by the National Natural Science Foundation of China(No.81473639)Beijing Natural Science Foundation(No.7122147)the Special Scientific Research Fund for Doctoral Subjects in Universities and Colleges(No.20121106110003)
文摘Objective:To investigate protective effects of hirudin on oxidative stress and apoptosis of spinal dorsal root ganglion cells in high-glucose rats at the cellular and molecular level.Methods:Dorsal root ganglion neurons(DRGn)were harvested from embryonic day in 15 SD rats,purified and identificated after primary culture.They were divided into the normal control group,high-glucose(HG)group,positive control(alpha-lipoic acid,ALA)group,low-dose hirudin group(H1),medium-dose hirudin group(H2)and high-dose hirudin group(H3).The control group was cultured by neuron specific culture medium,while the HG group was cultured by neuron specific culture medium and 20 mmol/L glucose(HG medium).The hirudin groups were cultured by HG medium+0.25 IU/mL hirudin(H1),HG medium+0.5 IU/mL hirudin(H2)and HG medium+1 IU/mL hirudin(H3).The ALA group was cultured by HG medium +100μmol/L ALA.3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylt etrazolium bromide(MTT)assay was used to explore the optimum concentration and intervention time.Flow cytometry assay was used to detect the level of reactive oxygen series(ROS).Western blot and quantificational realtime polymerase chain reaction(qRT-PCR)were used to detect the expression of protein and mRNA of nuclear factor erythroid 2-related factor 2(Nrf-2),hemeoxygence-1(HO-1),nuclear factor-κB(NF-κB)and Caspase-3.TUNEL assay was used to test the apoptosis rate of different groups.Results:After 24 h of culture,the cell activity of hirudin and ALA groups were higher than that of HG group,and there was a statistical difference between the H1 group and HG group(P<0.05).In hirudin groups,the apoptosis rate of cells,the expression of activated Caspase-3 protein and Caspase-3 mRNA were lower than those of HG group(P<0.01),higher than those of ALA group(P<0.01 or P<0.05).The ROS level of hirudin groups was higher than that of ALA group(P<0.01),lower than that of HG group(P<0.01 or P<0.05).The expression of NF-κB(P65)protein in H3 group were lower than those of HG group(P<0.05).The expression of Nrf-2 protein
基金the office of Traditional Chinese Medicine of Peking Union Medical College Hospitalthe Institute of Basic Medical Sciences of the Chinese Academy of Medical Sciences, Pecking, China, for supporting this research.
文摘Objective： TO examine the effects of the combination of quercetin （Q）, cinnamaldehyde （C） and hirudin （H）, a Chinese medicine formula on high glucose （HG）-induced apoptosis of cultured dorsal root ganglion （DRG） neurons. Methods： DRG neurons exposed to HG （45 mmol/L） for 24 h were employed as an in vitro model of diabetic neuropathy. Cell viability, reactive oxygen species （ROS） level and apoptosis were determined. The expression of nuclear factor of Kappa B （NF- κB）, inhibitory kappa B α （IκBα ）, phosphorylated IκBα and Nf-E2 related factor 2 （Nrf2） were examined using reverse transcription-polymerase chain reaction （RT-PCR） and Western blot assay. The expression of hemeoxygenase-1 （HO-1）, interleukin-6 （IL-6）, tumor necrosis factor （TNF-α） and caspase-3 were also examined by RT-PCR and Western blot assay. Results： HG treatment markedly increased DRG neuron apoptosis via increasing intracellular ROS level and activating the NF- κB signaling pathway （P〈0.05）. Co-treatment with Q, C, H and their combination decreased HG-induced caspase-3 activation and apoptosis （P〈0.05 or P〈0.01）. The expressions of NF- κB, IL-6 and TNF-α were down- regulated, and Nrf2/HO-1 expression was up-regulated （P〈0.05 or P〈0.01）. QCH has better effect in scavenging ROS, activating Nrf-2/HO-1, and down-regulating the NF- κB pathway than other treatment group. Conclusions： DRG neurons＇ apoptosis was increased in diabetic conditions, which was reduced by QCH formula treatment. The possible reason could be activating Nrf-2/HO-1 pathway, scavenging ROS, and inhibition of NF- κB activation. The effect of QCH combination was better than each monomer or the combination of the two monomers.
基金supported by a grant from the Education Department of Heilongjiang Province of China,No.12541398
文摘Astragaloside Ⅳ is the main active compound of Astragalus membranaceus. Astragaloside Ⅳ has strong anti-oxidative activities and protective effects against progression of peripheral neuropathy. In this study, we determined whether astragaloside Ⅳ protects retinal ganglion cells（RGC） from oxidative stress injury using the rat RGC-5 cell line. Hydrogen peroxide（H_2O_2） was used to induce oxidative stress injury, with the protective effect of astragaloside Ⅳ examined. Cell Counting Kit-8 and 4′,6-diamidino-2-phenylindole staining showed that astragaloside Ⅳ increased cell survival rate and decreased apoptotic cell number. Flow cytometry showed that astragaloside Ⅳ decreased H_2O_2-induced reactive oxygen species levels. While laser confocal microscopy showed that astragaloside Ⅳ inhibited the H_2O_2-induced decrease of mitochondrial membrane potential. Western blot assay showed that astragaloside Ⅳ reduced cytochrome c release induced by H_2O_2, inhibited Bax and caspase-3 expression, and increased Bcl-2 expression. Altogether, these results indicate that astragaloside Ⅳ has potential protective effects against H_2O_2-induced oxidative stress in retinal ganglion cells.
基金Supported by the National Natural Science Foundation of China(No.81473639)the Natural Science Foundation of Beijing(No.7122147)
文摘Objective： To examine the mechanism underlying the beneficial role of cinnamaldehyde on oxidative damage and apoptosis in high glucose （HG）-induced dorsal root ganglion （DRG） neurons in vitro. Methods： HG-treated DRG neurons were developed as an in vitro model of diabetic neuropathy. The neurons were randomly divided into five groups： the control group, the HG group and the HG groups treated with 25, 50 and 100 nmol/L cinnamaldehyde, respectively. Cell viability was examined by 3-（4,5-dimethylthiazol-2-yl）-2,5- diphenyltetrazolium bromide （MTT） assay and apoptosis rate was evaluated by the in situ TdT-mediated dUTP nick end labeling （TUNEL） assay. The intracellular level of reactive oxygen species （ROS） was measured with flow cytometry. Expression of nuclear factor-kappa B （NF-κ B）, inhibitor of κ B （I κ B）, phosphorylated I κ B （p-IκB）, tumor necrosis factor （TNF）-α, interleukin-6 （IL-6） and caspase-3 were determined by western blotting and real-time quantitative reverse transcription polymerase chain reaction （RT-PCR）. Expression of nuclear factor erythroid 2-related factor 2 （Nrf2） and hemeoxygenase-1 （HO-1） were also measured by western blotting. Results： Cinnamaldehyde reduced HG-induced loss of viability, apoptosis and intracellular generation of ROS in the DRG neurons via inhibiting NF- K B activity. The western blot assay results showed that the HG-induced elevated expressions of NF- κB, I κ B and p-I κ B were remarkably reduced by cinnamaldehyde treatment in a dose-dependent manner （P〈0.01）. The HG-induced over-expression of NF-κ B p65 mRNA was remarkably attenuated after cinnamaldehyde treatment in a dose-dependent manner （P〈0.01）. However, the expressions of Nrf2 and HO-1 were not upregulated. Treatment with cinnamaldehyde not only attenuated caspase-3 activation and the caspase cleavage cascade in DRG neurons, but also lowered the elevated IL-6, TNF-α, cyclo-oxygenase and inducible nitric oxide synthase levels, indi
