丹参酚酸B对高糖诱导雪旺细胞氧化损伤及凋亡的保护作用  

Protective effects of Salvianolic acid B on Schwann cells oxidative injury and apoptosis induced by high glucose

作　　者：孙连庆[1,2] 曹丽君[1] 王煊[2] 李晓瑾[2] 薛冰[2] 张婷婷[2] 曲玲[2] 陆菊明[2] 

机构地区：[1]西安交通大学第一附属医院,陕西西安710061 [2]中国人民解放军总医院,北京100853

出　　处：《时珍国医国药》2015年第7期1625-1628,共4页Lishizhen Medicine and Materia Medica Research

基　　金：中国博士后科学基金(No.20090461435);陕西省自然科学基金(2014JM4118);陕西省中医管理局中医药科研课题(13-LC088)

摘　　要：目的研究丹参酚酸B(Salvianolic acid B,Sal B)对高糖培养雪旺细胞氧化应激损伤及凋亡的保护作用。方法原代培养雪旺细胞,分为正常糖组(5.6mmol·L-1)、高糖组(50mmol·L-1)、渗透压对照组(5.6mmol·L-1葡萄糖+44.4mmol·L-1甘露醇)及高糖加不同浓度Sal B(25、50、100μmol·L-1)处理组。MTT法检测细胞活性,流式细胞术检测活性氧(ROS)水平及细胞凋亡,Elisa法检测8-羟基脱氧鸟苷(8-OHd G)含量。RT-PCR检测bcl-2及bax mRNA的表达;Western blot检测bcl-2、bax及caspase-3的表达。结果与正常组相比,高糖明显提高了雪旺细胞内ROS水平(P<0.01),增加了8-OHd G的生成(P<0.01),下调bcl-2蛋白及mRNA的表达(P<0.01),上调bax蛋白及mRNA的表达(P<0.01),促进了caspase-3的活化及细胞凋亡(P<0.01)。Sal B可以降低高糖所致的雪旺细胞内ROS及8-OHd G水平的增高(P<0.01),上调bcl-2蛋白及mRNA的表达(P<0.01),下调bax蛋白及其mRNA的表达(P<0.01),降低了caspase-3的活化(P<0.01),抑制了雪旺细胞凋亡。结论 Sal B可以抑制高糖所致的雪旺细胞氧化应激损伤及凋亡。Objective To investigate the inhibitory effects of Salvianolic acid B（ Sal B） on the high glucose- induced Schwann cells（ SCs） injury via oxidative stress.Methods SCs were primarily cultured and were treated in duplicate consistently with 5.6m M of glucose as the control（ con）,with 50 m M of glucose as high glucose（ HG）,with HG in the presence of 25μM,50μM,100μM Sal B for 48 hrs,respectively.The osmotic controls was that treating the cells with 5.6m M glucose plus 44.4m M mannitol alternatively.Intracellular ROS generation and apoptosis were detected by flow cytometry analysis and the concentration of 8- hydroxy- 2- deoxy Guanosine（ 8- OHd G） was detected by Elisa.RT- PCR was performed to analyze the expression levels of bax and bcl- 2.Western blot were performed to analyze the expression levels of some important proteins,such as bcl- 2,bax and caspase- 3.Results The relative levels of intracellular hydrogen peroxides and the concentrations of 8- OHd G showed a marked increase in SCs that were exposed to HG group compared with normal glucose exposure（ P〈0.01）.The apoptosis in the HG group was significantly increased than those in the control（ P〈0.01）.Compared to the control,treatment with HG up- regulated the Bax expression of protein and mRNA（ P〈0.01）,but down- regulated the Bcl- 2 expression of protein and mRNA（ P〈0.01）.In addition,treatment with HG increased the activation of caspase- 3 in SCs（ P〈0.01）.Treatment with Sal B inhibited the HG- induced oxidative stress by reducing ROS production,8- OHd G levels and apoptosis in SCs（ P〈0.01）.Furthermore,Sal B mitigated the HG- mediated up- regulation of Bax expression in SCs（ P〈0.01）.In addition,treatment with Sal B attenuated the HG- induced activation of caspase- 3 and up- regulated the bax expression（ P〈0.01）.Conclusion Sal B antagonized the HG- induced oxidative stress- related injury and apoptosis in SCs.

关 键 词：高糖 氧化应激 凋亡 雪旺细胞 丹参酚酸B 

分 类 号：R285.5[医药卫生—中药学]