摘要
目的研究bcl-2基因转染对保护体外培养缺血再灌注损伤的大鼠皮层神经细胞的影响。方法体外培养24h大鼠大脑皮层神经细胞,随机分为正常对照组(Control)、缺血再灌注组(Ischemia-reperfusion,I/R)和bcl-2基因转染组(bcl-2)。建立缺血再灌注模型,应用脂质体将bcl-2基因转入神经细胞。流式细胞术检测各组细胞凋亡率,MTT法测定细胞活力。Western-Blot法检测各组细胞bcl-2、葡萄糖调节蛋白78(Glucose regulated protein 78,GRP78)的蛋白含量。结果缺血再灌注组细胞凋亡率较对照组明显升高,bcl-2转染后细胞凋亡率显著下降,差异有统计学意义(P<0.05)。I/R组细胞活力较对照组明显下降,转染组细胞活力较I/R组显著升高,差异有统计学意义(P<0.05)。I/R组bcl-2比对照组明显表达减少,转染后细胞bcl-2蛋白高表达;I/R组细胞GRP78含量比对照组明显增多,bcl-2转染后GRP78显著下降。结论 bcl-2基因转染对缺血再灌注的鼠大脑神经元有明显保护作用,可能与其降低内质网相关性凋亡蛋白表达有关。
Objective To investigate the effects of bcl-2 gene on endoplasmic reticulum related apoptosis induced by ischemia reperfusion injury in rat cortical neurons in vitro. Methods Cerebral cortical neurons of neonatal rat were cultured in vitro and the ischemia-reperfusion injury cell models were established.Neurons were randomly divided into three groups：control group, I/R group and bcl-2 group.Liposomes were used to transfect bcl-2 gene. Apoptosis in rats was assessed by flow cytometry. Neuron viabilities were detected by MTT. Western-Blot was used to detect the protein contents of bcl-2 and GRP78. Results The apoptosis rate of of I/R group was significantly higher than that of the control group.The apoptosis in rats was significantly decreased in bcl-2 groups statistically significant（P〈0.05）.Neuron viability in I/R group was significantly lower than that of the control group, and the viability in bcl-2 group was significantly higher than that of the I/R group（P〈0.05）. Compared with control group, the protein expression of bcl-2 was small in I/R group. The bcl-2 content was significantly higher after transfection.The amount of GRP78 was less in control group, and more in I/R group. The GRP78 expressions were apparently reduced in bcl-2 group. Conclusion Bcl-2 gene has protective effects on rat cortical neurons injuryed by ischemia reperfusion injury in vitro,which may be associated with the reduction of E-R related apoptosis protein-GRP78 by bcl-2.
出处
《中国热带医学》
CAS
2014年第8期905-908,共4页
China Tropical Medicine
基金
海南省自然科学基金(No.309062)
海南省科技厅重点科技计划项目(No.ZDXM20100047)
关键词
bcl-2
基因转染
缺血再灌注
内质网应激
神经元
bcl-2
Gene transfeefion
Ischemia reperfusion injury
Endoplasmic retieulum
Neurons
