TRPC通道对PC12细胞缺氧缺糖再灌注后氧化损伤的保护作用  

Protective of Transient Receptor Potential Canonical(TRPC) on Oxygen-Glucose Deprivation/Reperfusion(OGD-R)-induced Injury in PC12 Cells

作　　者：王岩[1] 王莎莉[1] 赵香琴[1] 陈笛[1] 

机构地区：[1]重庆医科大学基础医学院,重庆400016

出　　处：《中国生物化学与分子生物学报》2014年第1期85-92,共8页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金资助项目(No.81100453/HO422)~~

摘　　要：瞬时受体电位C通道(transient receptor potential canonical,TRPC)属于瞬时受体电位(TRP)离子通道家族成员之一,与Ca2+调控及氧化应激关系密切.然而,TRPC通道在缺血缺氧性脑损伤中的作用仍有争议.本实验采用MTT法、台盼蓝排斥实验、LDH漏出实验联合Annexin/PI染色流式分析等显示,当采用通道阻断剂—SKF96365特异阻断TRPC通道时,PC12细胞对缺氧缺糖再灌注(oxygen-glucose deprivation/reperfusion,OGD-R)引起的损伤变得更敏感,细胞凋亡增加.尽管同时采用SKF96365、NMDA受体、AMPA受体和L-型钙通道阻断剂可引起细胞损伤和死亡减弱,但仍呈现明显的SKF96365浓度依懒性,提示TRPC通道参与细胞对缺氧缺糖再灌注耐受的调节,具有保护作用.钙指示剂Fluo-3AM荧光标记结合激光共聚焦显微镜分析显示,SKF96365可明显降低缺氧缺糖再灌注后细胞内的Ca2+浓度.总之,实验结果提示,TRPC通道对缺氧缺糖再灌注引起的细胞损伤和凋亡具有保护作用,其机制可能涉及TRPC通道对细胞内钙浓度的调节,详尽机制有待进一步研究.The transient receptor potential canonical （TRPC） is a subfamily of transient receptor potential （TRP） permeable to Ca2＋ , which is closely related to Ca2＋ regulation and oxidative stress. However,the role of TRPC in hypoxic ischemic brain damage is still controversial. Our data imply that TRPC can protect the PC12 cells from the injury induced by oxygen-glucose deprivation/reperfusion （OGD-R）. The MTT assay, trypan blue exclusion, lactate dehydrogenase （LDH） release, Annexin/PI （AV/PI） dye showed that PC12 cells became more sensitive of OGD-R induced injury,and obviously increased of cell apoptosis after treated with TRPC blocker SKF96365. Even though PC12 cells injury, cell death were weakened using the NMDA-receptor blocker,calcium channel blocker and AMPA-receptor blocker. These changes were still reflected in SKF96365 dose-dependent manner. The dada showed that TRPC had protective action on the regulation of resist OGD- R. The calcium indicator Fluo-3 AM in combination with laser confocal microscopy showed that SKF96365 could obviously reduced the intracellular Ca2＋ after OGD-R. In conclusion,TRPC can protect the PC12 cells from OGD-R and that the mechanism may be related to the regulation of intracellular free calcium concentration, and the detailed mechanism needs further research.

关 键 词：瞬时受体电位C通道 SKF96365 PC12细胞 缺氧缺糖再灌注 

分 类 号：R338.8[医药卫生—人体生理学]