目的:观察哺乳动物雷帕雷素靶蛋白(m TOR)依赖性自噬信号转导通路对HIV-1 gp120V3环介导的海马神经元L-型钙离子通道电流的影响。方法:取出生1 d内的乳鼠海马神经元原代培养7 d后用于实验。实验分为2个平行实验组,即空白对照组、gp120V3组、自噬阻断剂3-甲基腺嘌呤(3-MA)组和gp120V3+3-MA组;空白对照组、gp120V3组、自噬激动剂雷帕雷素(rapamycin)组和gp120V3+rapamycin组。以膜片钳技术记录海马神经元的L型钙离子通道电流。结果:gp120V3组及3-MA组与空白对照组比较,L型钙离子通道电流密度增大(P〈0.05);gp120V3+3-MA组与gp120V3组比较,L-型钙离子通道电流密度增大(P〈0.05);rapamycin组与空白对照组比较,L型钙离子通道电流密度减小(P〈0.05);gp120V3+rapamycin组与gp120V3组比较,L型钙离子通道电流密度减小(P〈0.05)。结论:从电生理角度证明,自噬可能参与了HIV-1gp120V3环介导的海马神经元L型钙离子通道电流的改变。
AIM: To observe the influence of autophagy on L-type calcium channel current induced by gp120V3 loop in hippocampal neurons. METHODS: Hippocampal neurons were exteriorized from the newborn rats within 1 d and primarily cultured for 7 d. The neurons were divided into 2 parallel groups,namely control group,gp120V3 group,autophagy inhibitor 3-MA group,gp120V3 + 3-MA group; control group,gp120V3 group,autophagy activator rapamycin group,gp120V3 + rapamycin group. Whole-cell pathch-clamp was used to record L-type calcium channel current. RESULTS:Compared with control group,the density of L-type calcium channel current increased in gp120V3 group and 3-MA group( P〈0. 05). Compared with gp120V3 group,the density of L-type calcium channel current increased in gp120V3 + 3-MA group( P〈0. 05). Compared with control group,the density of L-type calcium channel current decreased in rapamycin group( P〈0. 05). Compared with gp120V3 group,the density of L-type calcium channel current increased in gp120V3 + rapamycin group( P〈0. 05). CONCLUSION: Autophagy may be involved in the gp120V3 loop-mediated L-type calcium channels in hippocampus neurons.