HSP70被抗体封闭后对星形胶质细胞损伤后BDNF表达的影响

Effect of antibody-blocked HSP70 on the expression of BDNF in injured astrocytes

目的:探讨用抗体封闭热休克蛋白70(Heat shock protein70,HSP70)后对星形胶质细胞损伤后表达脑源性神经营养因子(Brain-derived neurotropic factor,BDNF)的影响。方法:原代培养并纯化星形胶质细胞,采用划痕损伤法构建细胞损伤模型,通过培养液内加入HSP70抗体来进行干预。在不同时间点对对照组和损伤干预组进行BDNF免疫细胞化学染色,观察细胞增生情况及BDNF的表达强度,并用RT-PCR检测BDNFmRNA的表达水平。结果:与对照组相比,干预组BDNF的表达明显减弱(P<0.05,P<0.01),GFAPmRNA的表达也明显降低(P<0.05,P<0.01)。结论:星形胶质细胞损伤后,HSP70可能具有促进星形胶质细胞表达BDNF的作用。

Objective：To study its effect on the expression of BDNF from injured astrocytes after HSP70 were blocked by HSP70-antibody. Methods：We performed scratch injuries in the cultured rat astrocytes,and added HSP70-antibody into the nutrient medium at once after injury（intervention group ）.Then , at 30min,24h,4d,7d after injury,the expression of brain-derived neurotropic factor（BDNF）was studied by immunocytochemical and reverse transcriptase-polymerase chain reaction（RT-PCR）.The immunocytochemical pictures were analyzed by image analysis system to abtain the average optical density of BDNF. Results： The expression of BDNF increased gradually with injury time.Compared with the control group,the average optical density of BDNF in the intervention group were significantly reduced（P〈0.05,P〈0.01 ）.The expresstion of BDNFmRNA was down-regulated in the intervention group（P〈0.05,P〈0.01）. Conclusion：HSP70 may play a role in the production of BDNF from injured astrocytes.