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作 者:刘阳生 陈晓云 邱堃[2] 王静[2] 姚华宁[2] 廖燕宏 Liu Yangsheng;ChenXiaoyun;Qiu Kun;Wang Jing;Yao Huaning;Liao Yanhong(Xianning First People’s Hospital,Xianning,437000;Wuhan First Hospital,Wuhan,430030;Tongji Medical College,Huazhong University of Science and Technology,Wuhan,430030)
机构地区:[1]咸宁市第一人民医院,咸宁437000 [2]武汉市第一医院,武汉430030 [3]华中科技大学同济医学院,武汉430030
出 处:《中国组织化学与细胞化学杂志》2021年第2期146-151,共6页Chinese Journal of Histochemistry and Cytochemistry
基 金:国家自然科学基金(31171087、30970662)。
摘 要:目的研究TRPC3/6/7在氧糖剥夺/复氧(oxygen glucose deprivation/reoxygenation,OGD/R)后对星形胶质细胞凋亡的影响及其可能机制。方法将体外培养星形胶质细胞分为空白对照组、野生型(wild type,WT)OGD/R组、TRPC3/6/7基因敲除OGD/R组,通过Western blot检测星形胶质细胞OGD/R后Bcl-2、Bax的变化,通过钙信号荧光测量技术检测星形胶质细胞OGD/R下的钙内流,通过流式细胞术对3组细胞进行凋亡率检测和分析。结果TRPC3/6/7基因敲除鼠星形胶质细胞OGD/R后Bcl-2水平较野生型OGD/R组明显增高,Bax水平较野生型OGD/R组明显降低;钙影像结果显示,TRPC3/6/7基因敲除的星形胶质细胞经OGD/R处理后,Tg刺激引起的钙内流较野生型OGD/R组明显降低;TRPC3/6/7基因敲除鼠星形胶质细胞OGD/R后凋亡率较野生型OGD/R组明显降低。结论TRPC3/6/7基因敲除可抑制OGD/R后星形胶质细胞凋亡的发生,并且能够抑制钙超载引起的脑缺血再灌注损伤,降低Ca2+稳态失衡在脑缺血再灌注脑损伤中的作用。Objective To investigate the effect of TRPC3/6/7 channel on apoptosis of astrocytes after oxygen-glucose deprivation/reoxygenation (OGD/R) and its possible mechanism.Methods The cerebral cortex astrocytes of neonatal mice were divided into control group,wild type OGD/R group,and TRPC3/6/7 knockout OGD/R group.Both OGD/R groups were cultured in the oxygen-glucose deprivation condition for a certain period to create the OGD/R model.The change of Bcl-2 and Bax protein expression in astrocytes was measured by Western blotting,the calcium influx in astrocytes was detected by calcium signal fluorescence measurement,and the apoptotic rate of astrocytes was analyzed by flow cytometry.Results After TRPC3/6/7 knockout,the level of Bcl-2 was significantly increased and the level of Bax was significantly decreased compared to the wild-type OGD/R group.Calcium imaging results showed the calcium influx in TRPC3/6/7 knockout astrocytes after OGD/R treatment was significantly lower than that of astrocytes in the wild-type OGD/R group.The apoptosis rate of astrocytes after TRPC3/6/7 knockout was significantly lower than that of astrocytes in the wild-type OGD/R group.Conclusion TRPC3/6/7 knockout can inhibit the apoptosis of astrocytes after OGD/R and inhibit cerebral ischemia/reperfusion injury caused by calcium overload.
分 类 号:R743.3[医药卫生—神经病学与精神病学]
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