三七皂苷R1对高糖诱导HK-2细胞转分化的作用及其机制研究

摘要

目的:探讨三七皂苷R1对高糖诱导人肾小管上皮细胞(HK-2)间质转分化(EMT)的作用及其可能机制.方法:传代培养HK-2细胞,并将细胞分为4组:对照组、高渗组、高糖组、高糖+三七皂苷R1组,均培养48 h.采用MTT法筛选药物浓度,western blotting法检测炎症相关蛋白NF-κB、肿瘤坏死因子-α(TNF-α)的表达,酶联免疫吸附试验(ELISA)法检测EMT标志性蛋白α平滑肌肌动蛋白(α-SMA)、人纤维连接蛋白(FN)的表达.结果:MTT显示60 μmol/L以下为三七皂苷R1的安全范围.高糖组、高糖+三七皂苷R1组NF-κB p65、TNF-α、α-SMA、FN蛋白表达均显著高于对照组和高渗组(P＜0.05),高渗组与对照组比较,差异无统计学意义(P＞0.05);与高糖组相比,高糖+三七皂苷R1组NF-κB p65、TNF-α、α-SMA、FN表达水平均明显降低(P＜0.05).结论:三七皂苷R1可抑制高糖诱导的HK-2细胞转分化,减轻炎性反应,延缓肾间质纤维化,其机制可能与抑制NF-κB/TNF-α信号通路有关.%Objective:To investigate the effect and mechanisms of Notoginsenoside R1 (NTR1) on high glucose induced epithelial-mesenchymal transition (EMT) in renal tubular epithelial cells (HK-2).Methods:HK-2 cells were divided into 4 groups:control group,mannitol group,high glucose group,and high glucose + NTR1 group.The cells were cultured for 48 h.The drug concentration was screened by MTT.The expressions of nuclear factor-κB (NF-κB) and tumor necrosis factor (TNF)-α were determined by western blotting.The levels of α-smooth muscle actin (α-SMA) and fibronectin (FN) were detected by enzyme linked immunosorbent assay (ELISA).Results:The safe dose of NTR1 was below 60 μmol/L.The protein expressions of NF-κB p65,TNF-α,α-SMA and FN in high glucose group and high glucose + NTR1 group were significantly higher than those in the control group and mannitol group (P ＜0.05),but no significant difference was found between the control group and mannitol group (P ＞ 0.05).Compared with the high glucose group,the expression of NF-κB p65,TNF-α,α-SMA and FN in high glucose + NTR1 group was notably decreased (P ＜0.05).Conclusion:NTR1 could inhibit high glucose induced EMT in HK-2 cells,alleviate inflammatory response,and attenuate renal interstitial fibrosis.The mechanisms might be related to the suppression of NF-κB/TNF-α signaling pathway.