糖络宁对高糖环境下雪旺细胞线粒体相关内质网膜的作用机制研究

Study on effect of Tangluoning on Schwann cells mitochondria-associated membranes in high glucose environment

目的:观察糖络宁含药血清对高糖环境下雪旺细胞(SCs)线粒体相关内质网膜(MAMs)的影响。方法:采用高糖环境培养48h的SCs(RSC96细胞株)模型,设立空白对照组、模型对照组、α硫辛酸(ALA)组、1%糖络宁组、10%糖络宁组。在电镜观察MAMs形态的基础上,采用免疫荧光标记法定量检测SCs中MAMs。梯度超速离心法分离MAMs,通过ELISA法检测MAMs上PKR样内质网激酶(PERK)的表达、Western Blot检测MAMs上线粒体融合蛋白2(Mfn2)的表达。结果:与空白对照组比较,模型对照组MAMs数量降低、PERK表达显著升高、Mfn2表达显著降低(P<0.01);与模型对照组比较,1%、10%糖络宁组MAMs数量均显著增加,PERK表达降低而Mfn2表达显著升高(P<0.01)。结论:糖络宁能够增加高糖环境下SCs中MAMs,与其增加Mfn2,降低PERK的表达有关。

Objective:To explore the effect of Tangluoning medicated serum in Schwann cells(SCs)mitochondriaassociated membranes(MAMs)incubated by high glucose.Methods:High glucose incubated SCs were used as cell models followed by setting control group,model group,Alpha lipoic acid(ALA)group,1%Tangluoning containing serum(LTLN)group and 10%Tangluoning containing serum(HTLN)group.Based on the observation of MAMs morphology by electron microscope,separation of MAMs,by gradient ultracentrifugation and detection of PKR-like endoplasmic reticulum kinase(PERK)expression on MAMs by ELISA.Western blot analysis was used to detect the expression of mitofusin 2(Mfn2)on MAMs in SCs.Results:Compared with control group,the number of MAMs decreased,the expression of PERK increased and the expression of Mfn2 decreased in model group(P<0.01).Compared with model group,the number of MAMs increased,the expression of PERK decreased and the expression of Mfn2 increased in LTLN group and HTLN group(P<0.01).Conclusion:Tangluoning can increase MAMs in SCs in high glucose environment,which is related to increasing Mfn2 and decreasing PERK expression.