甘木通乙酸乙酯提取物对低糖缺氧诱导的神经细胞的保护作用

目的 研究甘木通乙酸乙酯提取物对低糖缺氧诱导神经细胞凋亡的保护作用.方法 PC12细胞结合物理缺氧方式建立缺血性中风的细胞模型,CCK-8检测细胞活性,测定乳酸脱氢酶(LDH)漏出分析细胞膜完整性,流式细胞术和Hoechst 33258染色检测细胞凋亡,JC-1法测定细胞内线粒体膜电位,Western blot检测凋亡相关蛋白Bcl-2、Bax、cleaved caspase-3蛋白表达,检测SOD、MDA水平分析甘木通乙酸乙酯提取物的抗氧化能力.结果 与模型组比较,甘木通乙酸乙酯提取物可以有效提高缺氧PC12细胞的存活率(P<0.01),降低LDH漏出量(P<0.01),提高线粒体膜电位,增加细胞内SOD水平,降低MDA水平,增加Bcl-2蛋白表达,减少Bax,cleaved caspase-3蛋白的表达(P<0.05,P<0.01),降低细胞凋亡率.结论 甘木通乙酸乙酯提取物可抑制低糖缺氧诱导PC12细胞凋亡,该神经保护作用可能与细胞的线粒体凋亡途径有关.

Protective effects of ethyl acetate extract from Clematis filamentosa on nerve cells impaired by hypoglycemia and hypoxia

AIM To study the protective effects of ethyl acetate extract from Clematis filamentosa Dunn.(EAECFD)on nerve cells damaged by hypoglycemia and hypoxia.METHODS PC12 cell models of ischemic stroke were established with physical hypoxia.The cells were subjected to the detections of cellular activity by CCK 8,cell membrane integrity by lactate dehydrogenase(LDH)leakage,apoptosis by both flow cytometry and Hoechst 33258 staining,intracellular mitochondrial membrane potential by JC 1 method,apoptosis related proteins Bcl 2,Bax,and cleaved caspase 3 protein expressions by Western blot,and SOD and MDA levels as well to analyze the antioxidant capacity of ethyl acetate extract from C.filamentosa.RESULTS Compared with the model group,the group intervened with ethyl acetate extract from C.filamentosa displayed improved survival of hypoxic PC12 cells(P<0.01),less LDH leakage(P<0.01),increased mitochondrial membrane potential and intracellular SOD lev els,and reduced MDA,increased Bcl 2 protein expression,decreased Bax,cleaved caspase 3 protein expression(P<0.05,P<0.01),and inhibited apoptosis rate.CONCLUSION EAECFD can inhibit the apoptosis of PC12 cells damaged by hypoglycemia and hypoxia,and this neuroprotective effect may be related to the cellular mitochon drial apoptosis pathway.