黄芪注射液对高糖所致ECV304细胞损伤的保护作用

Protective effects of radix astragali on ECV304 cells exposed to high level of glucose

目的:探讨高糖对人脐静脉内皮细胞的损伤作用及其高糖导致细胞损伤的途径,为寻找出更为有效的糖尿病治疗途径提供理论依据。方法:将体外培养人脐静脉内皮细胞(ECV304细胞)分为高糖组(葡萄糖终浓度35mmol.L-1)、黄芪保护组(葡萄糖终浓度35mmol.L-1+黄芪终浓度500mg.L-1)、甘露醇高渗对照组(简称甘露醇组,甘露醇终浓度35mmol.L-1)和正常细胞组,在35mmol.L-1的高糖条件下处理ECV304细胞24h后检测细胞内Ca2+浓度、线粒体膜电位的测定并对不同处理组的细胞制作扫描电镜图片,观察细胞及线粒体形态的改变。结果:高糖组细胞内Ca2+浓度显著高于正常细胞组、黄芪保护组、甘露醇组(P<0.05);高糖组的线粒体膜电位显著低于正常细胞组、黄芪保护组、甘露醇组(P<0.05)。黄芪保护组和甘露醇组细胞内Ca2+浓度高于正常细胞组(P<0.05),线粒体的膜电位显著低于正常细胞组(P<0.05)。黄芪保护组细胞内的Ca2+浓度虽低于甘露醇组,但无显著性差异(P>0.05);黄芪保护组的线粒体膜电位显著地高于甘露醇组(P<0.05)。电镜下可见,黄芪保护组线粒体形态完好,线粒体内的嵴清晰可见,高糖组的线粒体出现肿胀,线粒体内的嵴消失。黄芪保护组可见到细胞的核分裂相,高糖组坏死细胞增多。甘露醇组细胞形态正常,线粒体的形态正常,线粒体内的嵴清晰可见。结论:黄芪对高糖导致的ECV304细胞损伤,尤其是线粒体损伤有较好的保护作用。

Objective To explore the damage effect of high level of glucose on ECV304 cells and the approaches in which high level of glucose plays its part to provide theoretical bases for the therapy of diabetes. Methods ECV304 cells were divided into four groups： normal control, high glucose group in which glucose was added to the cells with final concentration of 35 mmol·L^-1 , Radix Astragali （RA） group in which glucose was added to the cells with final concentration of 35 mmol·L^-1 , as well as RA with final concentration of 500 mg·L^-1 , mannitol group in which mannitol was added to the cells with final concentration of 25 mmol·L^-1. The cells were cultivated for 24 h after the glucose, RA and mannitol were added to the cells and collected for the determination of intracellular Ca^2＋ concentration, mitochondrial membrane potential and the morphological changes of cells and mitochondria were observed under electronic microscope. Results The intraeellular Ca^2＋ concentration in the cells of high glucose group was significantly higher than those of RA group, mannitol group and normal control group （P〈0.05）, the intracellular Ca^2＋ concentrations in the cells of RA group and mannitol group were significantly higher and the mitochondrial membrane potential was significantly lower than those of normal control group （P〈0.05）; the mitochondrial membrane potential in the cells of high glucose group was signifieantly lower than those of RA group, normal control group and mannitol group （P〈0.05）. The mitochondrial membrane potential in the cells of RA group was significantly higher than that of mannitol group （P〈0.05）. The morphology of cells in RA group was normal, the mitochondrial structure was normal, the cristae in mitoehondria were clear. The morphology of cells in high glucose group was abnormal, the mitochondria were swollen and the cristae were gone. Conclusion RA has protective effects on injury of ECV304 cells, especially mitochondrial injury, induced with high level of glucose.