氯化钴缺氧处理对大鼠星形胶质细胞TFPI-2表达的影响  

Effect of hypoxia on the expression of TFPI-2 in rat astrocytes induced by cobalt chloride

作　　者：孙佳[1] 高宏生[2] 王毅铮[2] 任贵兵[3] 孟斌[2] 龚燕华[4] 李灵芝 张永亮 钟士江[1] 

机构地区：[1]武警后勤学院附属医院脑科医院,天津300162 [2]武警后勤学院救援医学系卫生学教研室,天津300309 [3]武警后勤学院附属医院肿瘤外科,天津300162 [4]武警后勤学院临床医学系生物化学与分子生物学教研室,天津300309 [5]天津市职业与环境危害防控重点实验室,天津300162

出　　处：《武警后勤学院学报（医学版）》2017年第1期28-31,共4页Journal of Logistics University of PAP（Medical Sciences）

基　　金：国家自然科学基金资助项目(81273048);中国博士后基金(20100470106);武警后勤学院创新团队项目(WHTD201303);武警后勤学院附属医院种子基金项目(FYM201624)

摘　　要：【目的】探讨氯化钴缺氧对大鼠星型胶质细胞TFPI-2基因表达的影响及其可能的调节机制。【方法】将大鼠星型胶质细胞分为对照组、化学缺氧组(加入终浓度为100μmol/L氯化钴培养24 h)。显微镜下观察细胞形态变化,MTT法检测细胞活性,RT-PCR法检测各组缺氧诱导因子-1α(hypoxia induced factor,HIF-1α)、组织因子途径抑制因子-2(tissue factor pathway inhibitor 2,TFPI-2)mRNA表达水平变化,Western blotting检测缺氧前后HIF-1α、TFPI-2、VEGF蛋白表达水平变化。【结果】MTT检测显示,氯化钴缺氧会降低细胞活性,且呈时间依赖性。在显微镜下可观察到细胞缺氧后胞体变小,突触缩短。氯化钴缺氧处理组细胞TFPI-2 mRNA水平显著高于对照组(P<0.05)。Western blotting显示与对照组相比,缺氧后细胞内HIF-1α、TFPI-2、VEGF蛋白表达水平均有显著升高(P<0.05)。【结论】氯化钴缺氧可以激活HIF-1/VEGF缺氧通路,增加SH-SY5Y细胞TFPI-2 mRNA和蛋白表达水平。[Objective]To To investigate the effect of hypoxia on the expression of TFPI-2 in rat astrocytes induced by cobalt chloride and its potential mechanism. [ Methods] The rat astrocytes were randomly divided into two groups： control group and hypoxia group （cultured in 100 Ixmol/L COC12 for 24 h）. Then the cell morphology was observed under microscope and the cell viability was detected via MTT assay. RT-PCR was used to examine the mRNA expressions of HIF-1 c~ and TFP1-2. The protein expression levels of HIF-lc~, TFPI-2 and VEGF were assessed by Western blotting before and after hypoxia. [ Results] The results of MTT assay showed that the hypoxia induced by CoC12 decreased cell viability in a time-dependent manner. Under microscope, hypoxic ceils were observed with smaller cell bodies and shorter neurites. The mRNA level of TFPI-2 in the hypoxia group was significantly higher than that in the control group （P〈0.05）. The protein expression levels of HIF-1, TFPI-2 and VEGF in the hypoxia group were obviously higher than those in the control group （P〈0.05）. [ Conclusion ] Hypoxia induced by COC12 can activate HIF-1/VEGF hypoxia pathway, increase the mRNA and protein expressions of TFPI-2 in SH-SY5Y cells.

关 键 词：缺氧诱导因子-1Α 氯化钴 大鼠星型胶质细胞 组织因子途径抑制物-2 缺氧调控 

分 类 号：R743[医药卫生—神经病学与精神病学]