α-硫辛酸对波动性高糖诱导乳鼠雪旺细胞损伤的影响及机制  

Effect ofα-lipoic acid on intermittent high glucose-induced Schwann cell injury of suckling rats via mitochondrial pathway

作　　者：齐灵铄 李继扬 王倩倩 孙连庆[1] QI Lingshuo;LI Jiyang;WANG Qingqing;SUN Lianqing(The First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710061,China;不详)

机构地区：[1]西安交通大学第一附属医院,西安710061 [2]吴起县人民医院 [3]中国电子科技集团公司第二十研究所职工医院

出　　处：《山东医药》2020年第16期1-4,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81673785,81874447);陕西省中医药管理局项目(2019-ZZ-JC048)。

摘　　要：目的探讨α-硫辛酸(ALA)对波动性高糖诱导乳鼠雪旺细胞(SC)损伤的影响及机制。方法原代培养Wistar乳鼠SC,将其随机分组,正常对照组用5.6 mmol/L葡萄糖的DMEM培养基培养;稳定性高糖组用50 mmol/L葡萄糖的DMEM培养基培养;波动性高糖组用5.6 mmol/L葡萄糖的DMEM与50 mmol/L葡萄糖的DMEM培养基交替培养,每8 h更换培养液;低、中、高浓度ALA组在波动性高糖组基础上分别加50、100、200μmol/L ALA进行培养。均干预48 h收集各组细胞。用ELISA法检测细胞培养上清液中丙二醛(MDA)、超氧化物歧化酶(SOD),用流式细胞术检测线粒体膜电位(MMP),用TUNEL法测算细胞凋亡率,用Western blotting法检测细胞内B细胞淋巴瘤/白血病-2蛋白(Bcl-2)、Bcl-2相关X蛋白(bax)、活化型半胱氨酸天冬氨酸蛋白酶9(cleaved-caspase-9)、cleaved-caspase-3及线粒体、胞质内细胞色素C(cyto C)蛋白表达。结果与正常对照组、稳定性高糖组比较,波动性高糖组细胞培养液中MDA水平高,SOD水平低,SC凋亡率高,MMP低,Bcl-2蛋白相对表达量低,bax、cleaved-caspase-9、cleaved-caspase-3蛋白相对表达量高(P均<0.05);线粒体内cyto C蛋白表达低,而胞质中cyto C蛋白表达高(P均<0.05)。与波动性高糖组比较,低、中、高浓度ALA组培养液中MDA水平低,SOD水平高;MMP高;细胞内Bcl-2蛋白相对表达量高,bax、cleaved-caspase-9、cleaved-caspase-3蛋白相对表达量低(P均<0.05);线粒体cyto C蛋白表达高,胞质中cyto C蛋白表达低(P均<0.05),且以上指标变化呈浓度依赖性。结论ALA可通过线粒体途径抑制SC凋亡,对波动性高糖损伤的乳鼠SC有保护作用,且该作用呈浓度依赖性。Objective To investigate the effect ofα-lipoic acid(ALA)on the intermittent high glucose(IHG)-induced injury of Schwann cells(SCs)in the suckling rats via mitochondrial pathway.Methods SCs were obtained from the sciatic nerves of suckling rats.The cultured cells were treated for 48 hours with 5.6 mmol/L glucose-DMEM as control group(Con group),consistently with 50 mmol/L glucose-DMEM as the high glucose group HG group,with 5.6 mmol/L glucose-DMEM and 50mmol/L glucose-DMEM altering every 8 hours to mimic HG as the IHG group,with intermittent high glucose and 50,100,and 200μmol/L ALA,respectively,as the low-,medium-,and high-dose ALA groups.The concentration of malondialdehyde(MDA)and superoxide dismutase(SOD)activity were detected by ELISA,while the mitochondrial transmembrane potential(MMP)was detected by flow cytometry.The ratio of apoptopic cells were detected by TUNNEL method.The protein expression levels of Bcl-2,bax,cleaved-caspase-9,cleaved-caspase-3,and cyto C were detected by Western blotting.Results Compared with the Con group and HG group,MDA level increased,SOD activity decreased,SC apoptosis rate increased,MMP decreased,Bcl-2 protein expression decreased,bax,cleared-caspase-9,and cleared-caspase-3 proteins in cell culture medium increased in the IHG group(all P<0.05);the expression of cyto C protein in the mitochondria decreased,while the expression of cyto C protein in the cytoplasm increased(both P<0.05).Compared with the IHG group,the MDA level was lower in the culture medium,the SOD activity and MMP were higher,the relative expression of Bcl-2 protein in cells was higher,bax,cleared-caspase-9,and cleared-caspase-3 protein were lower,mitochondrial cyto C protein expression was higher,and cytoplasmic cyto C protein expression was lower in the cytoplasm in the low-,medium-,and high-dose ALA groups(all P<0.05);and the changes in the above indicators were in a dose-dependent manner.Conclusion ALA can inhibit the apoptosis of SCs of sucking rat via mitochondrial pathway,which has a significant protect

关 键 词：雪旺细胞损伤 波动性高糖 细胞凋亡 Α-硫辛酸 

分 类 号：R-332[医药卫生]