丹参酮ⅡA抑制波动性高糖诱导的雪旺细胞凋亡的观察

The protection of tanshinone ⅡA on schwann cells apoptosis induced by intermittent high glucose

目的观察丹参酮ⅡA（TanⅡA）对波动性高糖（IHG）所致的雪旺细胞（SC）凋亡的影响。方法原代培养大鼠SC分为正常糖组（Con,5.6 mmol/L）、稳定性高糖组（HG,50 mmol/L）、IHG组（IHG,5.6~50.0mmol/L）、渗透压对照组（mannitol）及IHG加不同浓度TanⅡA（0.1、1、10μmol/L）组。检测线粒体膜电位（MMP）、细胞凋亡率及凋亡相关蛋白表达。结果与Con组和HG组比较,IHG组MMP下降[（0.51±0.02）vs（1.42±0.08）vs（0.66±0.02）],细胞凋亡率增高[（5.01±0.26）%vs（42.33±2.76）%vs（31.64±3.04）%]（P〈0.05或P〈0.01）、线粒体细胞色素c（cyto-c）及凋亡诱导因子（AIF）水平降低,胞浆cyto-c及胞核AIF表达增高（P〈0.01）;活化半胱天冬氨酸蛋白酶-9（caspase-9）及caspase-3表达增高（P〈0.01）。TanⅡA可升高MMP,减少细胞凋亡,抑制cyto-C释放及AIF转位,降低caspase-9及caspase-3的活化。结论 TanⅡA能抑制SC凋亡,其机制是caspase活性降低,从caspase依赖性及非依赖性途径发挥作用。

Objective To investigate the influence of tanshinone ⅡA（TanⅡA）on Schwann cells（SCs）apoptosis induced by intermittent high glucose（IHG）. Methods SCs from primary cultured rats were divided into seven groups：SCs with normal glucose of 5.6 mmol/L（Con group）,SCs with high glucose of 50mmol/L（HG group）,SCs with intermittent high glucose of 5.6~50mmol/L（IHG group）,mannitol group,and IHG plus different concentrations of with Tan ⅡA（0.1,1,and 10μmol/L）groups.Mitochondrial transmembrane potential（MMP）,apoptosis rate and the expression levels of apoptosisrelated proteins were determined in all groups. Results As compared with Con and HG groups,MMP was lower[（0.51±0.02）vs（1.42±0.08）vs（0.66±0.02）],apoptosis rate was higher [（5.01±0.26）%vs（31.64±3.04）%vs（42.33±2.76）%]（P〈0.05 or P〈0.01）in IHG group.The expression levels of cytochrome c（cyto-c）and apoptosis inducing factor（AIF）in mitochondria decreased,but the expression levels of cytoplasmic cyto-c and nuclear AIF in cytoplasm and nucleus were increased（P〈0.01）in IHG group,and the expression levels of activated caspase-9and caspase-3were higher（P〈0.01）in IHG group.TanⅡA was likely to increase MMP,reduce cell,inhibit cyto-c release and AIF transposition and decrease caspase-9and caspase-3activation Conclusion TanⅡA may reduce the activity of caspase through caspase-dependent and-independent pathway,thereby protect SCs from apoptosis.