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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 protective effect of salvianolic acid B (Sal B) on oxidative stress injury and apoptosis in cultured Schwann cells cultured in high glucose medium. Methods Primary cultured Schwann cells were divided into three groups: normal glucose group (5.6 mmol·L-1), high glucose group (50 mmol·L-1), osmotic pressure control group (5.6 mmol·L-1 glucose + 44.4 mmol·L -1 mannitol) and high glucose plus Sal B (25,50,100μmol·L-1). Cell viability was measured by MTT assay, reactive oxygen species (ROS) levels and apoptosis were detected by flow cytometry. Elisa assay was used to detect the content of 8-OHdG. The expression of bcl-2 and bax mRNA was detected by RT-PCR. The expressions of bcl-2, bax and caspase-3 were detected by Western blot. Results Compared with normal group, high glucose significantly increased the level of ROS in Schwann cells (P <0.01), increased the production of 8-OHd G (P <0.01) and down-regulated the expression of bcl-2 protein and mRNA (P < 0.01), upregulated the protein and mRNA expression of bax (P <0.01), and promoted the activation and apoptosis of caspase-3 (P <0.01). Sal B could decrease the level of ROS and 8-OHd G in Schwann cells (P <0.01), up-regulate the expression of bcl-2 protein and mRNA (P <0.01) and down-regulate the protein and mRNA of bax (P <0.01), decreased the activation of caspase-3 (P <0.01), and inhibited the apoptosis of Schwann cells. Conclusion Sal B can inhibit oxidative stress injury and apoptosis of Schwann cells induced by high glucose.