Long-standing inflammation has emerged as a hallmark of neoplastic transformation of epithelial cells and may be a limiting factor of successful conventional tumor therapies.A complex milieu composed of distinct strom...Long-standing inflammation has emerged as a hallmark of neoplastic transformation of epithelial cells and may be a limiting factor of successful conventional tumor therapies.A complex milieu composed of distinct stromal and immune cells,soluble factors and inflammatory mediators plays a crucial role in supporting and promoting various types of cancers.An augmented inflammatory response can predispose a patient to colorectal cancer(CRC).Common risk factors associated with CRC development include diet and lifestyle,altered intestinal microbiota and commensals,and chronic inflammatory bowel diseases.Cysteinyl leukotrienes are potent inflammatory metabolites synthesized from arachidonic acid and have a broad range of functions involved in the etiology of various pathologies.This review discusses the important role of cysteinyl leukotriene signaling in linking inflammation and CRC.展开更多
Objective Aquaporin-4 （AQP4）, the main water channel protein in the brain, plays a critical role in water homeostasis and brain edema. Here, we investigated its role in the inflammatory responses after focal cerebra...Objective Aquaporin-4 （AQP4）, the main water channel protein in the brain, plays a critical role in water homeostasis and brain edema. Here, we investigated its role in the inflammatory responses after focal cerebral ischemia. Methods In AQP4-knockout （KO） and wild-type mice, focal cerebral ischemia was induced by 30 rain of middle cerebral arterial occlusion （MCAO）. Ischemic neuronal injury and cellular inflammatory responses, as well as the expression and localization of cysteinyl leukotriene CysLT2 and CysLT~ receptors, were determined at 24 and 72 h after MCAO. Results AQP4-KO mice showed more neuronal loss, more severe microglial activation and neutrophil infiltration, but less astrocyte proliferation in the brain after MCAO than wild-type mice. In addition, the protein levels of both CysLT1 and CysLT2 receptors were up-regulated in the ischemic brain, and the up-regulation was more pronounced in AQP4-KO mice. The CysLT1 and CysLT2 receptors were primarily localized in neurons, microglia and neutrophils; those localized in microglia and neutrophils were enhanced in AQP4-KO mice. Conclusion AQP4 may play an inhibitory role in postischemic inflammation.展开更多
Objective: To study the effect of Danshao Huaxian capsule (丹芍化纤胶囊,DSHX), a traditional Chinese medical prescription, on the expression of collagen (Col) Ⅰ, Ⅲ and cysteinyl aspartate specific proteases-3 (caspa...Objective: To study the effect of Danshao Huaxian capsule (丹芍化纤胶囊,DSHX), a traditional Chinese medical prescription, on the expression of collagen (Col) Ⅰ, Ⅲ and cysteinyl aspartate specific proteases-3 (caspase-3) in CCl4-induced hepatic fibrosis in rats. And also it is to explore the mechanism of DSHX in anti-fibrosis. Methods: Eighty male Wistar rats were randomly divided into the normal control group (A), the model group (B), the un-treated model group (C), the low-dose-DSHX treated group (D) and the high-dose-DSHX treated group (E). Except those in Group A, all the other rats were made into hepatic fibrotic models by comprehensive processes including subcutaneous injecting of CCl4, and feeding them with alcoholic high-fat and low-protein diet for 8 weeks. Then the two DSHX-treated groups were treated respectively with low dose (0.5 g/kg) and high dose (1.0 g/kg) DSHX capsule by gastrogavage everyday for 8 weeks. At the end of the experiment, the liver index, levels of hyaluronic acid (HA) and alanine amin-otransferase (ALT) in serum, degree of hepatic fibrosis, and urinary excretion of hydroxyproline (Hyp) were measured, and the expression of Col Ⅰ , Col Ⅲ and caspase-3 in liver tissues were detected respectively by immunohistochemistric technique. Results: Compared with those in Group B and C, the two DSHX treated groups showed that the liver index, levels of serum HA and ALT and severity of hepatic fibrosis were all significantly lower, the urinary excretion of Hyp was significantly higher; the Col Ⅰ and Col-Ⅲ expression was lower (Col Ⅰ :1. 23±1.14,1. 07±0. 96 vs 4.18±2. 26, 3. 22±1. 44, P<0. 01;Col Ⅲ : 1. 31±0. 69, , 1. 09± 0.58 vs 3.04±0.62,2.23±0.58, P<0.05). At the same time, the expression of caspase-3 in Group E was fewer than Group B and C in hepatocytes (3. 09±0. 65 vs 9. 60±2. 32, 8. 82 ±1. 45, P<0.01),but it was extensively expressed in fibrous septal cells(4.52±0.87 vs 1.69±0.23,2.98±0.36, P<0.01). Conclusion: DSHX capsule shows certain therapeutic effe展开更多
OBJECTIVE Previously we demonstrated the neuroprotective effect of 5-lipoxygenase(5-LOX)inhibitor as well as cysteinyl leukotriene receptor 1(Cys LT1)antagoniston rotenone-induced microglial activation and neuronal de...OBJECTIVE Previously we demonstrated the neuroprotective effect of 5-lipoxygenase(5-LOX)inhibitor as well as cysteinyl leukotriene receptor 1(Cys LT1)antagoniston rotenone-induced microglial activation and neuronal death.In this study,we determined the effects of 5-LOX inhibitor zileuton and Cys LT1 antagonist montelukast on neurotoxicity induced by 1-methyl-4-phenylpyridine(MPP+)in an in vitro model of Parkinson disease(PD).METHODS The neurotoxicity of MPP+,a neurotoxin relevant to PD,on the PC12 cells was measured by MTT assay,lactate dehydrogenase(LDH)release and double fluorescence staining with Hoechst/propidiumiodide(PI).The protective effects of 5-LOX inhibitor zileuton and Cys LT1 antagonist montelukast were investigated by the above methods.RESULTS We found that exposure of PC12 cells to MPP+led to a reduced cell viability and an increased level of LDH in a concentration-dependent manner.Pretreatment with zileuton and montelukast significantly attenuated viability loss and LDH release in MPP+-treated PC12 cells.Furthermore,MPP+increasednecrotic cell death in PC12 cells.Administration of montelukast significantly decreased MPP+-induced cell necrosis in PC12 cells.CONCLUSION The 5-LOX inhibitor zileuton and Cys LT1 antagonist montelukast have a neuroprotective effects on MPP+-induced neurotoxicity in PC12 cells.The 5-LOX inhibitor and Cys LT1 antagonist might raise a possibility as potential therapeutic agent for PD and other inflammation-related the central nervous system disorders.展开更多
The attachments of phycobilins to cysteinyl residues have been worked out through the reactions of phycoerytbrobilin dimethyl ester and phycocyanobilin dimethyl ester with cysteine methyl ester and reduced glutathione...The attachments of phycobilins to cysteinyl residues have been worked out through the reactions of phycoerytbrobilin dimethyl ester and phycocyanobilin dimethyl ester with cysteine methyl ester and reduced glutathione dimethyl ester respectively. A series of model compounds which carry the same conjugative skeletons as the chromophores in phycobillproteins have been purified anti identified. These compounds are characterized with strong fluorescence emission and circular dichroism effects which appeared weaker for the common bilinoid chromophores. Analyses of the circular dichroism effects of these compounds indicated that the major reaction products of phycobilins with cysteine methyl ester and reduced glutathione dimethyl ester carried the same stereochemical configuration as the chromophores in phvcobilinoroteins展开更多
OBJECTIVE We have recently reported that cysteinyl leukotriene(Cys LT) signaling plays an important role in microglial interleukin(IL)-1β secretion and subsequent neurotoxicity.The present study aimed to examine micr...OBJECTIVE We have recently reported that cysteinyl leukotriene(Cys LT) signaling plays an important role in microglial interleukin(IL)-1β secretion and subsequent neurotoxicity.The present study aimed to examine microglial morphological changes and the upstream molecular underlying IL^(-1)β production in Cys LT receptor agonist leukotriene D4(LTD4)-treated BV2 microglia in vitro.METHODS Twenty-four hours after murine microglial BV2 cells were stimulated with LTD4(1-100 nmol·L^(-1)),the cell proliferation and morphology were observed.The expression level of cysteinyl aspartate-specific protease 1(CASP1) protein was measured by Western blotin BV2 cells.In addition,BV2 cells were pretreated with or without CysLT1 receptor antagonist montelukast for 1 h and the effects of monte-lukaston LTD4-stimulated microglial activation and CASP1 expression were evaluated.RESULTS The number of BV2 cells had an increasing tendency after 24 h treatment with LTD4,but no significant differences were observed between the control and LTD4-treated cells(P>0.05).Under basal and resting conditions,BV2 microglial cells displayed a ramified morphology.However,LTD4 at 100 nmool·L^(-1) drove microglial morphological changes from a ramified towards an amoeboid shape.The expression of CASP1 protein was significantly upregulated in 100 nmool·L^(-1) LTD4-treated BV2 microglia(P<0.01).Furthermore,pretreatment with CysLT1 receptor antagonist montelukast prevented cell morphological changes and suppressed the increased CASP1 expression in LTD4-treated BV2 cells(P<0.05).CONCLUSION Cys LT receptor agonist LTD4 induces morphological changes and CASP1 expressionin BV2 microglia,which can be inhibited by CysLT1 antagonist.These results suggest the involvement of Cys LT signaling in microglial morphological changes and CASP1 expression.展开更多
Brian ischemic injury and central neurodegenerative diseases as leading contributors to disability and death have become a majorclinical and public health concern worldwide.Neuroinflammation plays a pivotal role in th...Brian ischemic injury and central neurodegenerative diseases as leading contributors to disability and death have become a majorclinical and public health concern worldwide.Neuroinflammation plays a pivotal role in the pathological progression of cerebral ischemia and neurodegenerative diseases including Parkinson disease(PD).Therefore,it is important to find effective therapeutic targets to attenuate inflammation and delay the progression of brain injury.Cysteinyl leukotrienes(CysLTs) are potent inflammatory mediators synthesized from arachidonic acid by 5-lipoxygenase(5-LOX) in the central nervous system.Two distinct G-protein-coupled receptors,CysLT1 R and CysLT2 R,mediate most of the known CysLTs biological responses.Accumulating evidence has demonstrated that postischemic inflammation and neuronal loss are mediated by 5-LOX and CysLTRs fol owing focal cerebral ischemia.We recently reported that the expression of 5-LOX,CysLT1R and inflammatory vascular cell adhesion molecule-1(VCAM-1) was upregulated in the hippocampus of rats with transient global cerebral ischemia,which was closely associated with delayed neuronal death in the hippocampal CA1 area.5-LOX inhibitor zileuton,CysLT1R antagonist ONO-1078 and montelukast dose-dependently reduced hippocampal CA1 neuronal death and inhibited the increased expression of 5-LOX and VCAM-1.In vitro ischemia-like injury in 5-LOXtransfected PC12 cells,oxygen-glucose deprivation(OGD) induced cell death mediated by5-LOX via ROS/P38 MAPK pathway.The nonselective 5-LOX inhibitor caffeic acid inhibited OGDstimulated activation of 5-LOX and ROS/P38 MAPK signaling and improved neuronal survival.In PD model,high concentrations of rotenone caused directly PC12 neurotoxicity,which was modulated by 5-LOX and abolished by suppression of 5-LOX.It is well known that microglia is major modulators of inflammatory response after brain injury.Overactivated microglia and production of proinflammatory cytokine IL-1β,IL-6 and TNF-α contribute to the neuroinflammation and brain injury.5-LOX展开更多
AIM： To investigate the expression and activity of leukotriene C4 （LTC4） synthesis enzymes and their underlying relationship with cysteinyl leukotriene （cys-LT） generation in a rat fulminant hepatic failure （FHF...AIM： To investigate the expression and activity of leukotriene C4 （LTC4） synthesis enzymes and their underlying relationship with cysteinyl leukotriene （cys-LT） generation in a rat fulminant hepatic failure （FHF） model induced by D-galactosamine/lipopolysaccharide （D-GaIN/ LPS）. METHODS： Rats were treated with D-GaIN （300 mg/kg） plus LPS （0.1 mg/kg） for 1, 3, 6, and 12 h. Enzyme immunoassay was used to determine the hepatic cys-LT content. Reverse transcription-polymerase chain reaction （RT-PCR）, Western blot or immunohistochemical assay were employed to assess the expression or location of LTC4 synthesis enzymes, which belong to membrane associated proteins in eicosanoid and glutathione （MAPEG） metabolism superfamily. Activity of LTC4 synthesis enzymes was evaluated by determination of the products of LTA4 after incubation with liver microsomes using high performance liquid chromatography （HPLC）. RESULTS： Livers were injured after treatment with D-GaIN/LPS, accompanied by cys-LT accumulation at the prophase of liver injury. Both LTC4 synthase （LTC4S） and microsomal glutathione-S-transferase （mGST） 2 were expressed in the rat liver, while the latter was specifically located in hepatocytes. Their mRNA and protein expressions were up-regulated at an earlier phase after treatment with D-GaIN/LPS. Meantime, a higher activity of LTC4 synthesis enzymes was detected, although theactivity of LTC4S played the main role in this case. CONCLUSION： The expression and activity of both LTC4S and mGST2 are up regulated in a rat FHF model, which are, at least, partly responsible for cys-LT hepatic accumulation.展开更多
It is well-established that following ingestion of aspirin or any other inhibitor of cyclooxygenase-1, patients with Samter’s disease, or aspirin-exacerbated respiratory disease (AERD) develop the sudden onset of wor...It is well-established that following ingestion of aspirin or any other inhibitor of cyclooxygenase-1, patients with Samter’s disease, or aspirin-exacerbated respiratory disease (AERD) develop the sudden onset of worsening respiratory clinical symptoms, which usually in-volves nasal congestion, rhinorrhea, wheezing and bronchospasm. Gastrointestinal distress, nausea, a pruritic rash and angioedema can also occasionally develop. However, the underlying pathologic mechanism that drives these clinical reactions remains elusive. Pretreatment with medications that inhibit the leukotriene pathway decreases the severity of clinical reactions, which points to the involvement of cysteinyl leukotrienes (cysLTs) in the pathogenesis of these aspirin-induced reactions. Furthermore, studies of aspirin challenges in carefully-phenotyped patients with AERD have confirmed that both proinflammatory lipid mediators, predominantly cysLTs and prostaglandin (PG) D 2, and the influx of effector cells to the respiratory tissue, contribute to symptom development during aspirin-induced reactions. Mast cells, which have been identified as the major cellular source of cysLTs and PGD 2, are likely to be major participants in the acute reactions, and are an attractive target for future pharmacotherapies in AERD. Although several recent studies support the role of platelets as inflammatory effector cells and as a source of cysLT overproduction in AERD, it is not yet clear whether platelet activation plays a direct role in the development of the aspirin-induced reactions. To further our understanding of the pathogenesis of aspirin-induced reactions in AERD, and to broaden the pharmacotherapeutic options available to these patients, additional investigations with targeted clinical trials will be required.展开更多
Objective:Tribulus terrestris saponin is a traditional Chinese medicine in China.This experiment was designed to investigate the effects of tribulus terrestris saponin on the proliferation and invasion ability of non-...Objective:Tribulus terrestris saponin is a traditional Chinese medicine in China.This experiment was designed to investigate the effects of tribulus terrestris saponin on the proliferation and invasion ability of non-small cell lung cancer A549 cells.Methods:A549 cells were divided into normal control and experimental groups(Tribulus terrestris saponin 250μg/mL group,Tribulus terrestris saponin 200μg/mL group,Tribulus terrestris saponin 150μg/mL group,Tribulus terrestris saponin 100μg/mL group,Tribulus terrestris saponin 50μg/mL group).The proliferation viability of the cells in each group was detected by CCK8,the invasion of tumor cells was detected by Transwell model.The mRNA expression of MMP9 and caspase-3 in each group of cells was detected by RT-PCR.Immunofluorescence staining was used to observe the fluorescence intensity of caspase-3 in each group of cells.Results:Compared with the normal control group,tribulus terrestris saponin significantly inhibited the proliferation activity and invasion ability of A549 cells,which was statistically significant(P<0.01).In the invasion assay,compared with the control group,MMP9 expression was significantly reduced and caspase-3 expression was significantly increased in the tribulus terrestris saponin group,and both were concentration-dependent,with statistically significant differences(P<0.01).By cellular immunofluorescence staining experiments,it was found that the fluorescence expression of caspase-3 was enhanced in the experimental group compared with the normal control group,in which the high concentration saponin group was significantly higher than the low concentration group.Conclusion:Tribulus terrestris saponin can inhibit the invasive ability of A549 cells by down-regulating the expression of MMP9,and induce irreversible apoptosis by up-regulating the activation of caspase-3 expression to form caspase-3.展开更多
文摘Long-standing inflammation has emerged as a hallmark of neoplastic transformation of epithelial cells and may be a limiting factor of successful conventional tumor therapies.A complex milieu composed of distinct stromal and immune cells,soluble factors and inflammatory mediators plays a crucial role in supporting and promoting various types of cancers.An augmented inflammatory response can predispose a patient to colorectal cancer(CRC).Common risk factors associated with CRC development include diet and lifestyle,altered intestinal microbiota and commensals,and chronic inflammatory bowel diseases.Cysteinyl leukotrienes are potent inflammatory metabolites synthesized from arachidonic acid and have a broad range of functions involved in the etiology of various pathologies.This review discusses the important role of cysteinyl leukotriene signaling in linking inflammation and CRC.
基金supported by the National Natural Science Foundation of China(81273491, 81072618,30772561 and 30873053)the Natural Science Foundation of Zhejiang Province,China(Y2090069)+1 种基金the"Qianjiang Rencai Research Plan"of Zhejiang Province China(2010R10055)the Fundamental Research Funds for the Central Universities,China(2009QNA7008)
文摘Objective Aquaporin-4 （AQP4）, the main water channel protein in the brain, plays a critical role in water homeostasis and brain edema. Here, we investigated its role in the inflammatory responses after focal cerebral ischemia. Methods In AQP4-knockout （KO） and wild-type mice, focal cerebral ischemia was induced by 30 rain of middle cerebral arterial occlusion （MCAO）. Ischemic neuronal injury and cellular inflammatory responses, as well as the expression and localization of cysteinyl leukotriene CysLT2 and CysLT~ receptors, were determined at 24 and 72 h after MCAO. Results AQP4-KO mice showed more neuronal loss, more severe microglial activation and neutrophil infiltration, but less astrocyte proliferation in the brain after MCAO than wild-type mice. In addition, the protein levels of both CysLT1 and CysLT2 receptors were up-regulated in the ischemic brain, and the up-regulation was more pronounced in AQP4-KO mice. The CysLT1 and CysLT2 receptors were primarily localized in neurons, microglia and neutrophils; those localized in microglia and neutrophils were enhanced in AQP4-KO mice. Conclusion AQP4 may play an inhibitory role in postischemic inflammation.
文摘Objective: To study the effect of Danshao Huaxian capsule (丹芍化纤胶囊,DSHX), a traditional Chinese medical prescription, on the expression of collagen (Col) Ⅰ, Ⅲ and cysteinyl aspartate specific proteases-3 (caspase-3) in CCl4-induced hepatic fibrosis in rats. And also it is to explore the mechanism of DSHX in anti-fibrosis. Methods: Eighty male Wistar rats were randomly divided into the normal control group (A), the model group (B), the un-treated model group (C), the low-dose-DSHX treated group (D) and the high-dose-DSHX treated group (E). Except those in Group A, all the other rats were made into hepatic fibrotic models by comprehensive processes including subcutaneous injecting of CCl4, and feeding them with alcoholic high-fat and low-protein diet for 8 weeks. Then the two DSHX-treated groups were treated respectively with low dose (0.5 g/kg) and high dose (1.0 g/kg) DSHX capsule by gastrogavage everyday for 8 weeks. At the end of the experiment, the liver index, levels of hyaluronic acid (HA) and alanine amin-otransferase (ALT) in serum, degree of hepatic fibrosis, and urinary excretion of hydroxyproline (Hyp) were measured, and the expression of Col Ⅰ , Col Ⅲ and caspase-3 in liver tissues were detected respectively by immunohistochemistric technique. Results: Compared with those in Group B and C, the two DSHX treated groups showed that the liver index, levels of serum HA and ALT and severity of hepatic fibrosis were all significantly lower, the urinary excretion of Hyp was significantly higher; the Col Ⅰ and Col-Ⅲ expression was lower (Col Ⅰ :1. 23±1.14,1. 07±0. 96 vs 4.18±2. 26, 3. 22±1. 44, P<0. 01;Col Ⅲ : 1. 31±0. 69, , 1. 09± 0.58 vs 3.04±0.62,2.23±0.58, P<0.05). At the same time, the expression of caspase-3 in Group E was fewer than Group B and C in hepatocytes (3. 09±0. 65 vs 9. 60±2. 32, 8. 82 ±1. 45, P<0.01),but it was extensively expressed in fibrous septal cells(4.52±0.87 vs 1.69±0.23,2.98±0.36, P<0.01). Conclusion: DSHX capsule shows certain therapeutic effe
基金The project supported National Natural Science Foundation of China(81273491)the Zhejiang Provincial Natural Science Foundation(LY12H31010)
文摘OBJECTIVE Previously we demonstrated the neuroprotective effect of 5-lipoxygenase(5-LOX)inhibitor as well as cysteinyl leukotriene receptor 1(Cys LT1)antagoniston rotenone-induced microglial activation and neuronal death.In this study,we determined the effects of 5-LOX inhibitor zileuton and Cys LT1 antagonist montelukast on neurotoxicity induced by 1-methyl-4-phenylpyridine(MPP+)in an in vitro model of Parkinson disease(PD).METHODS The neurotoxicity of MPP+,a neurotoxin relevant to PD,on the PC12 cells was measured by MTT assay,lactate dehydrogenase(LDH)release and double fluorescence staining with Hoechst/propidiumiodide(PI).The protective effects of 5-LOX inhibitor zileuton and Cys LT1 antagonist montelukast were investigated by the above methods.RESULTS We found that exposure of PC12 cells to MPP+led to a reduced cell viability and an increased level of LDH in a concentration-dependent manner.Pretreatment with zileuton and montelukast significantly attenuated viability loss and LDH release in MPP+-treated PC12 cells.Furthermore,MPP+increasednecrotic cell death in PC12 cells.Administration of montelukast significantly decreased MPP+-induced cell necrosis in PC12 cells.CONCLUSION The 5-LOX inhibitor zileuton and Cys LT1 antagonist montelukast have a neuroprotective effects on MPP+-induced neurotoxicity in PC12 cells.The 5-LOX inhibitor and Cys LT1 antagonist might raise a possibility as potential therapeutic agent for PD and other inflammation-related the central nervous system disorders.
基金This project was supported by the National Natural Science Foundation of China and the United Analytical Testing Center in Zhongguancun Area.
文摘The attachments of phycobilins to cysteinyl residues have been worked out through the reactions of phycoerytbrobilin dimethyl ester and phycocyanobilin dimethyl ester with cysteine methyl ester and reduced glutathione dimethyl ester respectively. A series of model compounds which carry the same conjugative skeletons as the chromophores in phycobillproteins have been purified anti identified. These compounds are characterized with strong fluorescence emission and circular dichroism effects which appeared weaker for the common bilinoid chromophores. Analyses of the circular dichroism effects of these compounds indicated that the major reaction products of phycobilins with cysteine methyl ester and reduced glutathione dimethyl ester carried the same stereochemical configuration as the chromophores in phvcobilinoroteins
基金supported by National Natural Science Foundation of China(81671188)Zhejiang Provincial Natural Science Foundation of China(LY12H31010)the Key Laboratory of Hangzhou City Project(20090233T12)
文摘OBJECTIVE We have recently reported that cysteinyl leukotriene(Cys LT) signaling plays an important role in microglial interleukin(IL)-1β secretion and subsequent neurotoxicity.The present study aimed to examine microglial morphological changes and the upstream molecular underlying IL^(-1)β production in Cys LT receptor agonist leukotriene D4(LTD4)-treated BV2 microglia in vitro.METHODS Twenty-four hours after murine microglial BV2 cells were stimulated with LTD4(1-100 nmol·L^(-1)),the cell proliferation and morphology were observed.The expression level of cysteinyl aspartate-specific protease 1(CASP1) protein was measured by Western blotin BV2 cells.In addition,BV2 cells were pretreated with or without CysLT1 receptor antagonist montelukast for 1 h and the effects of monte-lukaston LTD4-stimulated microglial activation and CASP1 expression were evaluated.RESULTS The number of BV2 cells had an increasing tendency after 24 h treatment with LTD4,but no significant differences were observed between the control and LTD4-treated cells(P>0.05).Under basal and resting conditions,BV2 microglial cells displayed a ramified morphology.However,LTD4 at 100 nmool·L^(-1) drove microglial morphological changes from a ramified towards an amoeboid shape.The expression of CASP1 protein was significantly upregulated in 100 nmool·L^(-1) LTD4-treated BV2 microglia(P<0.01).Furthermore,pretreatment with CysLT1 receptor antagonist montelukast prevented cell morphological changes and suppressed the increased CASP1 expression in LTD4-treated BV2 cells(P<0.05).CONCLUSION Cys LT receptor agonist LTD4 induces morphological changes and CASP1 expressionin BV2 microglia,which can be inhibited by CysLT1 antagonist.These results suggest the involvement of Cys LT signaling in microglial morphological changes and CASP1 expression.
基金The project supported by National Natural Science Foundation of China(81671188)Zhejiang Provincial Natural Science Foundation of China(LY12H31010)Key Laboratory of Hangzhou City Project(20090233T12)
文摘Brian ischemic injury and central neurodegenerative diseases as leading contributors to disability and death have become a majorclinical and public health concern worldwide.Neuroinflammation plays a pivotal role in the pathological progression of cerebral ischemia and neurodegenerative diseases including Parkinson disease(PD).Therefore,it is important to find effective therapeutic targets to attenuate inflammation and delay the progression of brain injury.Cysteinyl leukotrienes(CysLTs) are potent inflammatory mediators synthesized from arachidonic acid by 5-lipoxygenase(5-LOX) in the central nervous system.Two distinct G-protein-coupled receptors,CysLT1 R and CysLT2 R,mediate most of the known CysLTs biological responses.Accumulating evidence has demonstrated that postischemic inflammation and neuronal loss are mediated by 5-LOX and CysLTRs fol owing focal cerebral ischemia.We recently reported that the expression of 5-LOX,CysLT1R and inflammatory vascular cell adhesion molecule-1(VCAM-1) was upregulated in the hippocampus of rats with transient global cerebral ischemia,which was closely associated with delayed neuronal death in the hippocampal CA1 area.5-LOX inhibitor zileuton,CysLT1R antagonist ONO-1078 and montelukast dose-dependently reduced hippocampal CA1 neuronal death and inhibited the increased expression of 5-LOX and VCAM-1.In vitro ischemia-like injury in 5-LOXtransfected PC12 cells,oxygen-glucose deprivation(OGD) induced cell death mediated by5-LOX via ROS/P38 MAPK pathway.The nonselective 5-LOX inhibitor caffeic acid inhibited OGDstimulated activation of 5-LOX and ROS/P38 MAPK signaling and improved neuronal survival.In PD model,high concentrations of rotenone caused directly PC12 neurotoxicity,which was modulated by 5-LOX and abolished by suppression of 5-LOX.It is well known that microglia is major modulators of inflammatory response after brain injury.Overactivated microglia and production of proinflammatory cytokine IL-1β,IL-6 and TNF-α contribute to the neuroinflammation and brain injury.5-LOX
基金Supported by The National Natural Science Foundation of China, No. 30672564, No. 30472112 and No. 30070904
文摘AIM： To investigate the expression and activity of leukotriene C4 （LTC4） synthesis enzymes and their underlying relationship with cysteinyl leukotriene （cys-LT） generation in a rat fulminant hepatic failure （FHF） model induced by D-galactosamine/lipopolysaccharide （D-GaIN/ LPS）. METHODS： Rats were treated with D-GaIN （300 mg/kg） plus LPS （0.1 mg/kg） for 1, 3, 6, and 12 h. Enzyme immunoassay was used to determine the hepatic cys-LT content. Reverse transcription-polymerase chain reaction （RT-PCR）, Western blot or immunohistochemical assay were employed to assess the expression or location of LTC4 synthesis enzymes, which belong to membrane associated proteins in eicosanoid and glutathione （MAPEG） metabolism superfamily. Activity of LTC4 synthesis enzymes was evaluated by determination of the products of LTA4 after incubation with liver microsomes using high performance liquid chromatography （HPLC）. RESULTS： Livers were injured after treatment with D-GaIN/LPS, accompanied by cys-LT accumulation at the prophase of liver injury. Both LTC4 synthase （LTC4S） and microsomal glutathione-S-transferase （mGST） 2 were expressed in the rat liver, while the latter was specifically located in hepatocytes. Their mRNA and protein expressions were up-regulated at an earlier phase after treatment with D-GaIN/LPS. Meantime, a higher activity of LTC4 synthesis enzymes was detected, although theactivity of LTC4S played the main role in this case. CONCLUSION： The expression and activity of both LTC4S and mGST2 are up regulated in a rat FHF model, which are, at least, partly responsible for cys-LT hepatic accumulation.
基金This work was supported by the National Institutes of Health(NIH grant#K23HL111113 and#R01HL128241)by generous contributions from the Vinik and Kaye Families.
文摘It is well-established that following ingestion of aspirin or any other inhibitor of cyclooxygenase-1, patients with Samter’s disease, or aspirin-exacerbated respiratory disease (AERD) develop the sudden onset of worsening respiratory clinical symptoms, which usually in-volves nasal congestion, rhinorrhea, wheezing and bronchospasm. Gastrointestinal distress, nausea, a pruritic rash and angioedema can also occasionally develop. However, the underlying pathologic mechanism that drives these clinical reactions remains elusive. Pretreatment with medications that inhibit the leukotriene pathway decreases the severity of clinical reactions, which points to the involvement of cysteinyl leukotrienes (cysLTs) in the pathogenesis of these aspirin-induced reactions. Furthermore, studies of aspirin challenges in carefully-phenotyped patients with AERD have confirmed that both proinflammatory lipid mediators, predominantly cysLTs and prostaglandin (PG) D 2, and the influx of effector cells to the respiratory tissue, contribute to symptom development during aspirin-induced reactions. Mast cells, which have been identified as the major cellular source of cysLTs and PGD 2, are likely to be major participants in the acute reactions, and are an attractive target for future pharmacotherapies in AERD. Although several recent studies support the role of platelets as inflammatory effector cells and as a source of cysLT overproduction in AERD, it is not yet clear whether platelet activation plays a direct role in the development of the aspirin-induced reactions. To further our understanding of the pathogenesis of aspirin-induced reactions in AERD, and to broaden the pharmacotherapeutic options available to these patients, additional investigations with targeted clinical trials will be required.
基金National Key R&D Plan(2022YFC2305004)Hainan Province Major Science and Technology Special Project(No.ZDKJ2021036)+3 种基金Key R&D projects in Hainan Province(No.ZDYF2020223)Hainan Province Key R&D Plan International Science and Technology Cooperation Project(GHYF2022011)Hainan Provincial Innovation Team Project(No.820CXTD448)National Natural Science Foundation of China(No.82260001,82160012)。
文摘Objective:Tribulus terrestris saponin is a traditional Chinese medicine in China.This experiment was designed to investigate the effects of tribulus terrestris saponin on the proliferation and invasion ability of non-small cell lung cancer A549 cells.Methods:A549 cells were divided into normal control and experimental groups(Tribulus terrestris saponin 250μg/mL group,Tribulus terrestris saponin 200μg/mL group,Tribulus terrestris saponin 150μg/mL group,Tribulus terrestris saponin 100μg/mL group,Tribulus terrestris saponin 50μg/mL group).The proliferation viability of the cells in each group was detected by CCK8,the invasion of tumor cells was detected by Transwell model.The mRNA expression of MMP9 and caspase-3 in each group of cells was detected by RT-PCR.Immunofluorescence staining was used to observe the fluorescence intensity of caspase-3 in each group of cells.Results:Compared with the normal control group,tribulus terrestris saponin significantly inhibited the proliferation activity and invasion ability of A549 cells,which was statistically significant(P<0.01).In the invasion assay,compared with the control group,MMP9 expression was significantly reduced and caspase-3 expression was significantly increased in the tribulus terrestris saponin group,and both were concentration-dependent,with statistically significant differences(P<0.01).By cellular immunofluorescence staining experiments,it was found that the fluorescence expression of caspase-3 was enhanced in the experimental group compared with the normal control group,in which the high concentration saponin group was significantly higher than the low concentration group.Conclusion:Tribulus terrestris saponin can inhibit the invasive ability of A549 cells by down-regulating the expression of MMP9,and induce irreversible apoptosis by up-regulating the activation of caspase-3 expression to form caspase-3.
