α硫辛酸对高糖培养神经膜细胞氧化应激及凋亡的影响  

Effects of alpha lipoic acid on apoptosis and oxidative stress of Schwann cell induced by high glucose in vitro

作　　者：秦利[1] 刘志民[2] 

机构地区：[1]上海交通大学医学院附属新华医院内分泌科,上海200092 [2]上海长征医院内分泌科

出　　处：《上海医学》2013年第5期433-437,I0002,共6页Shanghai Medical Journal

基　　金：上海市科学技术委员会自然科学基金资助项目(10ZR1439400)

摘　　要：目的探讨α硫辛酸(ALA)对高糖培养氧化应激及凋亡的影响。方法原代培养并纯化Sprague-Dawley乳鼠坐骨神经神经膜细胞,应用S-100蛋白免疫组织化学法进行鉴定。取第3～4代神经膜细胞为研究对象,分为正常组(5.6mmol/L葡萄糖培养基)和高糖组(50mmol/L葡萄糖培养基),以及在高糖组内分别加入10、100和500μmol/LALA的高糖10ALA组、高糖100ALA组、高糖500ALA组。干预48h后采用四甲基偶氮唑盐微量酶反应比色法检测细胞活力,脱氧核苷酸末端转移酶介导的生物素化dUTP缺口末端标记技术检测细胞凋亡,Western印迹法检测B细胞淋巴瘤/白血病相关x蛋白(Bax蛋白)的表达,同时检测各组神经膜细胞的丙二醛(MDA)含量及超氧化物歧化酶(SOD)活性。结果高糖组、高糖10ALA组、高糖100ALA组、高糖500ALA组的神经膜细胞存活率均显著低于对照组(P值均<0.05),神经膜细胞凋亡率均显著高于对照组(P值均<0.05);高糖10ALA组、高糖100ALA组、高糖500ALA组的神经膜细胞存活率均显著高于高糖组(P值均<0.05),神经膜细胞凋亡率均显著低于高糖组(P值均<0.05)。高糖组的神经膜细胞MDA生成显著多于对照组(P<0.05),SOD活性显著低于对照组(P<0.05)。高糖10ALA组、高糖100ALA组、高糖500ALA组的神经膜细胞MDA生成均显著多于对照组(P值均<0.05),但均显著少于高糖组(P值均<0.05);SOD活性均显著低于对照组(P值均<0.05),但均显著高于高糖组(P值均<0.05)。高糖组的神经膜细胞Bax蛋白表达显著高于对照组(P<0.05)。高糖10ALA组、高糖100ALA组、高糖500ALA组的神经膜细胞Bax蛋白表达均显著高于对照组(P值均<0.05),但均显著低于高糖组(P值均<0.05)。结论 ALA通过抑制氧化应激而改善高糖导致的神经膜细胞凋亡,该机制可能在改善糖尿病周围神经病变中起重要作用。Objective To investigate the effects of alpha lipoic acid （ALA） on apoptosis and oxidative stress of Schwann cells cultured with high glucose medium in vitro. Methods Schwann cells were primarily cultured and purified from the sciatic nerves of newborn Sprague-Dawley rats. S-100 protein immunohistochemical method was adopted for the identification. Schwann cells of the 3rd and 4th passage were collected and divided into normal group （5.6 mmol/L glucose） and high glucose group （50 mmol/L glucose）. Various concentrations of ALA （10, 100 and 500 μmol/L） were added into high glucose medium to establish high glucose＋ 10 ALA group, high glucose＋ 100 ALA group, and high glucose＋500 ALA group, respectively. After 48-hour intervention, methyl thiazolyl tetrazolium （MTT） was adopted to detect the activity of Schwann cells. Apoptosis was confirmed by the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling （TUNEL）. Western blot was performed to detect the expression of B cell lymphoma/leukemia associationed x protein （Bax protein）. In addition, the oxidative stress indices in Schwann cells, such as the content of malondialdehyde （MDA） and the activity of superoxide dismutase （SOD） were also measured. Results The survival rates of Schwann cells in high glucosegroup, high glucose ＋ 10 ALA group, high glucose ＋ 100 ALA group, and high glucose ＋ 500 ALA group were significantly lower than the control group （all P〈0. 05）, while the proportion of cell apoptosis in high glucose groups was significantly higher than that in the control group （P〈0.05）. The survival rates of Schwann cells in high glucose ＋ 10 ALA group, high glucose＋ 100 ALA group and high glucose＋ 500 ALA group were significantly higher than high glucose group （all P〈0. 05）, while the proportion of cell apoptosis was significantly lower than high glucose group （all P〈0. 05）. MDA level in high glucose group was significantly higher than the control group （P〈0.05）, but

关 键 词：糖尿病周围神经病变 神经膜细胞 氧化应激 Α硫辛酸 

分 类 号：R587.2[医药卫生—内分泌]