杜仲多糖抑制高糖诱导HK⁃2细胞损伤的研究

Experimental study of Eeucommia polysaccharide on HK⁃2 cell damage induced by high glucose

目的探讨杜仲多糖(EP)对肾小管上皮细胞损伤的影响及分子机制。方法将人肾小管上皮细胞HK-2随机分为正常对照(NC)组、高糖组(HG),HG+EP 10组、HG+EP 20组、HG+EP 40组、HG+anti-miR-NC组、HG+anti-miR-1207-5p组、HG+miR-1207-5p组、HG+EP+miR-1207-5p组。细胞计数试剂盒8法检测细胞活性,流式细胞术检测细胞凋亡,ELISA法检测TNF-α、IL-6含量,实时荧光定量PCR检测miR-1207-5p表达。结果HG+EP 10、HG+EP 20、HG+EP 40组细胞活性依次升高(P<0.05),细胞凋亡率、Cleaved-caspase3蛋白表达、IL-6、TNF-α含量、miR-1207-5p表达依次降低(P<0.05)。与HG组比较,HG+miR-1207-5p组miR-1207-5p、细胞凋亡率及Cleaved-caspase3蛋白表达、IL-6、TNF-α升高(P<0.05),细胞活性降低(P<0.05)。与HG+EP 40组比较,HG+EP+miR-1207-5p组miR-1207-5p、细胞凋亡率、Cleaved-caspase3蛋白表达、IL-6、TNF-α升高(P<0.05),细胞活性降低(P<0.05)。结论EP通过下调miR-1207-5p表达抑制肾小管上皮细胞损伤。

Objective To explore the influence of Eeucommia polysaccharide on renal tubular epithelial cell injury and its molecular mechanism.Methods Human renal tubular epithelial cells HK-2 were randomly divided into normal control group(NC),high glucose group(HG),HG+EP 10 group,HG+EP 20 group,HG+anti-miR-NC group,HG+anti-miR-1207-5p group,HG+EP group,HG+miR-1207-5p group and HG+EP+miR-1207-5p group.Cell counting kit 8(CCK-8)method was used to detect cell activity and flow cytometry was used to detect cell apoptosis.Protein expression was evaluated by western blot method.The content of TNF-αand IL-6 was tested by ELISA.The expression level of miR-1207-5p was measured by Real-time fluorescent quantitative PCR(qRT-PCR).Results In HG+EP 10,HG+EP 20 and HG+EP 40 groups,the cell activity increased in turn(P<0.05),while the cell apoptosis rate,the expression of Cleaved-caspase3 protein,the expression of IL-6,TNF-αand miR-1207-5p decreased in turn(P<0.05).Compared with HG group,the expression of miR-1207-5p,the apoptosis rate,the expression of Cleavedcaspase 3 protein,the contents of IL-6 and TNF-αincreased(P<0.05),while the cell activity decreased in HG+miR-1207-5p group(P<0.05).Compared with HG+EP 40 group,miR-1207-5p,the apoptosis rate,the expression of Cleaved-caspase3 protein,the contents of IL-6 and TNF-αincreased(P<0.05),while the cell activity decreased in HG+EP+miR-1207-5p group(P<0.05).Conclusion Eucommia polysaccharide inhibits renal tubular epithelial cell damage by down-regulating the expression of miR-1207-5p.