摘要
目的 评价异氟醚后处理对大鼠缺氧缺糖-复氧复糖皮质神经元促凋亡蛋白表达的影响.方法 体外培养出生24 h内的SD大鼠原代皮质神经元,以1×106个/ml的密度接种于6孔板（2ml/孔）培养皿,采用随机数字表法分为3组（n=12）：对照组（C组）神经元正常培养;缺氧缺糖-复氧复糖组（OGD/R组）神经元缺氧缺糖30 min后复糖复氧;异氟醚后处理组（Ⅰ组）神经元缺氧缺糖30 min后复糖复氧即刻向培养皿中通入2％异氟醚,培养1h.于培养24h时收集神经元,采用Hoechst/PI双染色法测定神经元凋亡率;采用免疫印迹法测定cmpme-3蛋白的表达;采用PCR法测定Bid、B im和Puma的mRNA表达;采用Westem blot法测定Bid 、Bim和Puma的蛋白表达.结果 与S组比较,OGD/R组神经元凋亡率升高,caspase-3蛋白、Bid、Bim和Puma的mRNA和蛋白表达上调（P＜0.05）;与OGD/R组比较,Ⅰ组神经元凋亡率下降,caspase-3蛋白、Bid、Bim和Puma的mRNA和蛋白表达下调（P＜0.05）.结论 异氟醚后处理通过下调促凋亡蛋白表达,抑制大鼠缺氧缺糖-复氧复糖皮质神经元凋亡.
Objective To evaluate the effect of isoflurane post-conditioningon the expression of pro apoptotic proteins in the conical neurons exposed to oxygen-glucose deprivation and restoration （OGD/R） in rats.Methods Primary cortical neurons isolated from male Sprague-Dawley rats （within 24h after birth）,were cultured in vitro and inoculated in 6-well culture plate （2 ml/well） at a density of 1 × 10^6/ml.The cells were divided into 3 groups （n =12 each） using a random number table：control group（group C）,OGD/R group,and isoflurane post-conditioning group （Ⅰ group）.In OGD/R group,the cells were incubated in glucose-free BBS aerated with 95 % N2 for 30 min followed by restoration of 2-glucose supply for 1 h.At 24 h of incubation,the cells were collected for detection of neuronal apoptosis （.using Hoechst/PI staining）,caspase-3 expression （by Western blot）,expression of Bid,Bim and Puma mRNA （by PCR）,and expression of Bid,Bim and Puma （by Western blot）.Apoptosis rate was calculated.Results Compared with S group,the apoptosis rate was significantly increased,and the expression of caspase-3 and Bid,Bim and Puma mRNA and protein was upregulated in OGD/R group.The apoptosis rate and expression of caspase-3 and Bid,Bim and Puma mRNA and protein were significantly lower in Ⅰ group than in OGD/R group.Conclusion Isofluranepost-conditioning inhibits apoptosis in the cortical neurons exposed to OGD/R through down-regulating pm-apoptotic proteins in rats.
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2014年第12期1495-1497,共3页
Chinese Journal of Anesthesiology
关键词
异氟醚
缺血后处理
缺氧
氧
大脑皮质
神经元
凋亡调节蛋白质类
Isoflurane
Ischemic postconditioning
Anoxia
Oxygen
Cerebral cortex
Neurons
Apoptosis regulatory proteins
