Acupuncture has been shown to ameliorate cognitive impairment of Alzheimer’s disease.Acupoints and stimulation frequency influence the therapeutic effect of electroacupuncture.Rat models of Alzheimer’s disease were ...Acupuncture has been shown to ameliorate cognitive impairment of Alzheimer’s disease.Acupoints and stimulation frequency influence the therapeutic effect of electroacupuncture.Rat models of Alzheimer’s disease were established by injecting amyloid beta 1–42（Aβ_（1–42））into the bilateral lateral ventricles.Electroacupuncture at 2,30,and 50 Hz was carried out at Baihui（GV20;15°obliquely to a depth of 2mm）and Shenshu（BL23;perpendicularly to 4–6 mm depth）,once a day for 20 minutes（each）,for 15 days,taking a break every 7 days.The Morris water maze test was conducted to assess the learning and memory.The expression levels of glycogen synthase kinase-3β（GSK-3β）,p Ser9-GSK-3β,p Tyr216-GSK-3β,amyloid precursor protein and Aβ_（1–40） in the hippocampus were determined by western blot assay.Results demonstrated that electroacupuncture treatment at different frequencies markedly improved learning and memory ability,increased synaptic curvatures,decreased the width of synaptic clefts,thickened postsynaptic densities,and downregulated the expression of GSK-3β,amyloid precursor protein,and Aβ_（1–40）.pSer9-GSK-3βexpression markedly decreased,while p Tyr216-GSK-3βexpression increased.High-frequency（50 Hz）electroacupuncture was more effective than low（2 Hz）or medium-frequency（30 Hz）electroacupuncture.In conclusion,electroacupuncture treatment exerts a protective effect against Aβ_（1–42）-induced learning and memory deficits and synapse-ultrastructure impairment via inhibition of GSK-3βactivity.Moreover,high-frequency electroacupuncture was the most effective therapy.展开更多
Background Neuropathologically, Alzheimer disease （AD） is characterized by the presence of extracellular plaques enriched in β-amyloid peptides; however, the mechanism by which it results in the neurotoxicity is un...Background Neuropathologically, Alzheimer disease （AD） is characterized by the presence of extracellular plaques enriched in β-amyloid peptides; however, the mechanism by which it results in the neurotoxicity is uncertain. The purpose of this study was to investigate whether it would prompt the progress of Alzheimer disease via enhancement of aberrant phosphorylated tau that results from its increased kinase gene expression. Methods Twenty-four male rats were divided into three groups, and each group had 8 rats： control, sham-operated, and Aβ25-35 injected AD model groups. AD rat models were created by unilateral injections of Aβ25-35 into the amygdala. The hyperphosphorylated tau protein was estimated by immunohistochemistry with paired helical filament-1 （PHF-1） antibody and paired helical filament-tau （AT8） antibody. The expressions of glycogen synthase kinase-3β （GSK-3β） and p38 mitogen-activated protein kinase （P38MAPK） mRNA were observed by in situ hybridization. Results Compared with the control and sham-operated groups, the evaluation of paired AT8 and paired helical filament-1 （PHF-1） in the cortexes and hippocampus of the AD model group showed the numbers of AT8 and PHF-1 positive cells, as well as the optical density （OD） values of the proteins were significantly higher （AT8： in CA2： 0.318±0.037 vs. 0.135±0.028, 0.136±0.031; in frontal cortex： 0.278±0.040 vs. 0.130±0.028, 0.190±0.037. PHF-1 ： in CA2： 0.386±0.034 vs. 0.139±0.010, 0.193±0.041; in frontal cortex： 0.395±0.050 vs. 0.159±0.030, 0.190±0.044, respectively, P 〈0.01）; the number of GSK-3β mRNA and P38MAPK mRNA positive cells of the AD model group, as well as the OD values, also increased significantly in the cortexes, hippocampus （GSK-3β-mRNA： in CA2：0.384±0.012 vs. 0.190±0.015, 0.258±0.064; in frontal cortex： 0.398±0.018 vs. 0.184±0.031, 0.218±0.049. P38MAPK mRNA： in CA2：0.409±0.038 vs. 0.161±0.041, 0.189±0.035; in frontal cortex： 0.423±0.070 vs. 0.160±0展开更多
AIM： To investigate the regulatory mechanism of glycogen synthase kinase 3β（GSK3β） in epithelialmesenchymal transition（EMT） process after proliferative vitreoretinopathy（PVR） induction. METHODS： Experimenta...AIM： To investigate the regulatory mechanism of glycogen synthase kinase 3β（GSK3β） in epithelialmesenchymal transition（EMT） process after proliferative vitreoretinopathy（PVR） induction. METHODS： Experimental PVR was induced by intravitreal injection of retinal pigment epithelium（RPE） cells in the eyes of rabbits. A PI3 K/Akt inhibitor（wortmannin） and a GSK3β inhibitor（Li Cl） were also injected at different time during PVR progress. Electroretinogram（ERG）, ocular fundus photographs, and B-scan ultrasonography were used to observe the PVR progress. Western blot test on the extracted retina were performed at 1, 2, 4 wk. The expression of the mesenchymal marker vimentin was determined by immunohistochemistry. Toxicity of wortmannin and Li Cl were evaluated by ERG and Td Tmediated d UTP nick-end labeling（TUNEL） assay. The vitreous was also collected for metabolomic analysis. RESULTS： Experimental PVR could significantly lead to EMT, along with the suppressed expression of GSK3β and the activation of Wnt/β-catenin and PI3 K/Akt pathways. It was verified that upregulating the expression of GSK3β could effectively inhibit EMT process by suppressing Wnt/β-catenin and PI3 K/Akt pathways. CONCLUSION： GSK3β effectively inhibits EMT via the Wnt/β-catenin and PI3 K/Akt pathways. GSK3β may be regarded as a promising target of experimental PVR inhibition.展开更多
Background： Cognitive impairment is a severe complication caused by obstructive sleep apnea （OSA）. The mechanisms of causation are still unclear. The Wntβ-catenin signaling pathway is involved in cognition, and ab...Background： Cognitive impairment is a severe complication caused by obstructive sleep apnea （OSA）. The mechanisms of causation are still unclear. The Wntβ-catenin signaling pathway is involved in cognition, and abnormalities in it are implicated in neurological disorders. Here, we explored the Wntβ-catenin signaling pathway abnormalities caused by chronic intermittent hypoxia （CIH）, the most characteristic pathophysiological component of OSA. Methods： We divided 32 4-week-old male C57/BL mice into four groups of eight each： a CIH ＋ normal saline （NS） group, CIH ＋ LiCI group, sham CIH ＋ NS group, and a sham CIH ＋ LiCI group. The spatial learning performance of each group was assessed by using the Morris water maze （MWM）. Protein expressions of glycogen synthase kinase-β （GSK-β） and β-catenin in the hippocampus were examined using the Western blotting test. EdU labeling and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling staining methods were used, respectively, to determine the proliferation and apoptosis of neurons in the hippocampal dentate gyrus region. Results： Mice exposed to CIH showed impaired spatial learning performance in the MWM, including increased mean escape latencies to reach the target platform, decreased mean times passing through the target platform and mean duration in the target quadrant. The GSK-313 activity increased, and expression of β-catenin decreased significantly in the hippocampus of the CIH-exposed mice. Besides, CIH significantly increased hippocampal neuronal apoptosis, with an elevated apoptosis index. Meanwhile, LiCI decreased the activity of GSK-β and increased the expression of β-catenin and partially reversed the spatial memory deficits in MWM and the apoptosis caused by CIH. Conclusions： Wntβ-catenin signaling pathway abnormalities possibly play an important role in the development of cognitive deficits among mice exposed to CIH and that LiCI might attenuate CIH-induced cognitive impairment via Wntβ-c展开更多
As a major microtubule-associated protein, tau plays an important role in promoting microtubule assembly and stabilizing microtubules. In Alzheimer’s disease(AD) and other tauopathies, the abnormally hyperphosphoryla...As a major microtubule-associated protein, tau plays an important role in promoting microtubule assembly and stabilizing microtubules. In Alzheimer’s disease(AD) and other tauopathies, the abnormally hyperphosphorylated tau proteins are aggregated into paired helical filaments and accumulated in the neurons with the form of neurofibrillary tangles. An imbalanced regulation in protein kinases and protein phosphatases is the direct cause of tau hyperphosphorylation. Among various kinases and phosphatases, glycogen synthase kinase-3β(GSK-3β) and protein phosphatase 2A(PP2A) are the most implicated. Accumulation of the hyperphosphorylated tau induces synaptic toxicity and cognitive impairments. Here, we review the upstream factors or pathways that can regulate GSK-3β or PP2A activity mainly based on our recent findings. We will also discuss the mechanisms that may underlie tau-induced synaptic toxicity.展开更多
文摘目的分析糖尿病肾病患者血清糖原合酶激酶-3β(glycogen synthase kinase-3β,GSK-3β)水平与尿微量白蛋白的相关性,以探究GSK-3β与糖尿病肾病发生发展的关系。方法选取2016年2月至2019年11月新乡医学院第一附属医院收治的117例2型糖尿病患者,根据24h尿白蛋白排泄量(urinary albumin excretion,UAE),将其分为尿白蛋白正常组(UAE<30mg/24h)46例、微量白蛋白尿组(30mg/24h≤UAE≤300mg/24h)39例及大量白蛋白尿组(UAE>300mg/24h)32例,另将同期在本院体检的42例健康志愿者作为对照组。收集各组的临床资料,采用免疫比浊法测定尿液中白蛋白含量,采用酶联免疫吸附法检测血清中GSK-3β水平。结果与对照组相比,尿白蛋白正常组、微量白蛋白尿组、大量白蛋白尿组血清GSK-3β水平显著升高(P<0.05);与尿白蛋白正常组相比,微量白蛋白尿组、大量白蛋白尿组血清GSK-3β水平显著升高(P<0.05);与微量白蛋白尿组相比,大量白蛋白尿组血清GSK-3β水平显著升高(P<0.05)。在2型糖尿病患者中,血清GSK-3β水平与糖化血红蛋白、总胆固醇、低密度脂蛋白、甘油三酯、血肌酐、血尿酸、UAE、病程呈正相关(P<0.05),与高密度脂蛋白呈负相关(P<0.05)。多元逐步线性回归分析结果显示,糖化血红蛋白、血肌酐、血尿酸、UAE影响GSK-3β水平的独立危险因素(P<0.05)。结论血清GSK-3β水平与2型糖尿病患者尿微量白蛋白呈正相关,并随糖尿病肾病的发生及疾病严重程度的加重而逐渐升高,临床监测血清GSK-3β水平变化对于早期发现糖尿病肾病具有重要意义。
基金supported by the National Natural Science Foundation of China,No.81373741a grant from the Chinese Medicine and Integrated Medicine Research Projects funded by the Health and Family Planning Commission of Hubei Province of China,No.24a grant from the Hubei Provincial Collaborative Innovation Center of Preventive Treatment by Acupuncture and Moxibustion of China in 2014,No.8
文摘Acupuncture has been shown to ameliorate cognitive impairment of Alzheimer’s disease.Acupoints and stimulation frequency influence the therapeutic effect of electroacupuncture.Rat models of Alzheimer’s disease were established by injecting amyloid beta 1–42（Aβ_（1–42））into the bilateral lateral ventricles.Electroacupuncture at 2,30,and 50 Hz was carried out at Baihui（GV20;15°obliquely to a depth of 2mm）and Shenshu（BL23;perpendicularly to 4–6 mm depth）,once a day for 20 minutes（each）,for 15 days,taking a break every 7 days.The Morris water maze test was conducted to assess the learning and memory.The expression levels of glycogen synthase kinase-3β（GSK-3β）,p Ser9-GSK-3β,p Tyr216-GSK-3β,amyloid precursor protein and Aβ_（1–40） in the hippocampus were determined by western blot assay.Results demonstrated that electroacupuncture treatment at different frequencies markedly improved learning and memory ability,increased synaptic curvatures,decreased the width of synaptic clefts,thickened postsynaptic densities,and downregulated the expression of GSK-3β,amyloid precursor protein,and Aβ_（1–40）.pSer9-GSK-3βexpression markedly decreased,while p Tyr216-GSK-3βexpression increased.High-frequency（50 Hz）electroacupuncture was more effective than low（2 Hz）or medium-frequency（30 Hz）electroacupuncture.In conclusion,electroacupuncture treatment exerts a protective effect against Aβ_（1–42）-induced learning and memory deficits and synapse-ultrastructure impairment via inhibition of GSK-3βactivity.Moreover,high-frequency electroacupuncture was the most effective therapy.
基金This study was supported by the grants from the National Natural Science Foundation of China （No. 30971029）, Fund of Shanghai Science and Technology Committee （No. 08411950900）, and Young Excellent Talents Award of Tongji University （No. 2006KJ067）.
文摘Background Neuropathologically, Alzheimer disease （AD） is characterized by the presence of extracellular plaques enriched in β-amyloid peptides; however, the mechanism by which it results in the neurotoxicity is uncertain. The purpose of this study was to investigate whether it would prompt the progress of Alzheimer disease via enhancement of aberrant phosphorylated tau that results from its increased kinase gene expression. Methods Twenty-four male rats were divided into three groups, and each group had 8 rats： control, sham-operated, and Aβ25-35 injected AD model groups. AD rat models were created by unilateral injections of Aβ25-35 into the amygdala. The hyperphosphorylated tau protein was estimated by immunohistochemistry with paired helical filament-1 （PHF-1） antibody and paired helical filament-tau （AT8） antibody. The expressions of glycogen synthase kinase-3β （GSK-3β） and p38 mitogen-activated protein kinase （P38MAPK） mRNA were observed by in situ hybridization. Results Compared with the control and sham-operated groups, the evaluation of paired AT8 and paired helical filament-1 （PHF-1） in the cortexes and hippocampus of the AD model group showed the numbers of AT8 and PHF-1 positive cells, as well as the optical density （OD） values of the proteins were significantly higher （AT8： in CA2： 0.318±0.037 vs. 0.135±0.028, 0.136±0.031; in frontal cortex： 0.278±0.040 vs. 0.130±0.028, 0.190±0.037. PHF-1 ： in CA2： 0.386±0.034 vs. 0.139±0.010, 0.193±0.041; in frontal cortex： 0.395±0.050 vs. 0.159±0.030, 0.190±0.044, respectively, P 〈0.01）; the number of GSK-3β mRNA and P38MAPK mRNA positive cells of the AD model group, as well as the OD values, also increased significantly in the cortexes, hippocampus （GSK-3β-mRNA： in CA2：0.384±0.012 vs. 0.190±0.015, 0.258±0.064; in frontal cortex： 0.398±0.018 vs. 0.184±0.031, 0.218±0.049. P38MAPK mRNA： in CA2：0.409±0.038 vs. 0.161±0.041, 0.189±0.035; in frontal cortex： 0.423±0.070 vs. 0.160±0
基金Supported by the National Natural Science Foundation of China(No.81371039)Shanghai Natural Science Foundation(No.18ZR1440200)
文摘AIM： To investigate the regulatory mechanism of glycogen synthase kinase 3β（GSK3β） in epithelialmesenchymal transition（EMT） process after proliferative vitreoretinopathy（PVR） induction. METHODS： Experimental PVR was induced by intravitreal injection of retinal pigment epithelium（RPE） cells in the eyes of rabbits. A PI3 K/Akt inhibitor（wortmannin） and a GSK3β inhibitor（Li Cl） were also injected at different time during PVR progress. Electroretinogram（ERG）, ocular fundus photographs, and B-scan ultrasonography were used to observe the PVR progress. Western blot test on the extracted retina were performed at 1, 2, 4 wk. The expression of the mesenchymal marker vimentin was determined by immunohistochemistry. Toxicity of wortmannin and Li Cl were evaluated by ERG and Td Tmediated d UTP nick-end labeling（TUNEL） assay. The vitreous was also collected for metabolomic analysis. RESULTS： Experimental PVR could significantly lead to EMT, along with the suppressed expression of GSK3β and the activation of Wnt/β-catenin and PI3 K/Akt pathways. It was verified that upregulating the expression of GSK3β could effectively inhibit EMT process by suppressing Wnt/β-catenin and PI3 K/Akt pathways. CONCLUSION： GSK3β effectively inhibits EMT via the Wnt/β-catenin and PI3 K/Akt pathways. GSK3β may be regarded as a promising target of experimental PVR inhibition.
基金This work was supported by a grant from the National Natural Science Foundation of China （No. 81370185）.
文摘Background： Cognitive impairment is a severe complication caused by obstructive sleep apnea （OSA）. The mechanisms of causation are still unclear. The Wntβ-catenin signaling pathway is involved in cognition, and abnormalities in it are implicated in neurological disorders. Here, we explored the Wntβ-catenin signaling pathway abnormalities caused by chronic intermittent hypoxia （CIH）, the most characteristic pathophysiological component of OSA. Methods： We divided 32 4-week-old male C57/BL mice into four groups of eight each： a CIH ＋ normal saline （NS） group, CIH ＋ LiCI group, sham CIH ＋ NS group, and a sham CIH ＋ LiCI group. The spatial learning performance of each group was assessed by using the Morris water maze （MWM）. Protein expressions of glycogen synthase kinase-β （GSK-β） and β-catenin in the hippocampus were examined using the Western blotting test. EdU labeling and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling staining methods were used, respectively, to determine the proliferation and apoptosis of neurons in the hippocampal dentate gyrus region. Results： Mice exposed to CIH showed impaired spatial learning performance in the MWM, including increased mean escape latencies to reach the target platform, decreased mean times passing through the target platform and mean duration in the target quadrant. The GSK-313 activity increased, and expression of β-catenin decreased significantly in the hippocampus of the CIH-exposed mice. Besides, CIH significantly increased hippocampal neuronal apoptosis, with an elevated apoptosis index. Meanwhile, LiCI decreased the activity of GSK-β and increased the expression of β-catenin and partially reversed the spatial memory deficits in MWM and the apoptosis caused by CIH. Conclusions： Wntβ-catenin signaling pathway abnormalities possibly play an important role in the development of cognitive deficits among mice exposed to CIH and that LiCI might attenuate CIH-induced cognitive impairment via Wntβ-c
文摘As a major microtubule-associated protein, tau plays an important role in promoting microtubule assembly and stabilizing microtubules. In Alzheimer’s disease(AD) and other tauopathies, the abnormally hyperphosphorylated tau proteins are aggregated into paired helical filaments and accumulated in the neurons with the form of neurofibrillary tangles. An imbalanced regulation in protein kinases and protein phosphatases is the direct cause of tau hyperphosphorylation. Among various kinases and phosphatases, glycogen synthase kinase-3β(GSK-3β) and protein phosphatase 2A(PP2A) are the most implicated. Accumulation of the hyperphosphorylated tau induces synaptic toxicity and cognitive impairments. Here, we review the upstream factors or pathways that can regulate GSK-3β or PP2A activity mainly based on our recent findings. We will also discuss the mechanisms that may underlie tau-induced synaptic toxicity.
