小檗碱调节ERK/NF-κB信号通路改善高糖诱导的系膜细胞异常增殖

Berberine regulates ERK/NF-κB signaling pathway to improve high glucose-induced mesangial cell proliferation

目的探讨小檗碱(BBR)对高糖(HG)诱导的系膜细胞(MCs)异常增殖的调节作用及可能的机制。方法体外培养MCs,实验分为对照、高糖模型和BBR给药组(30、60、90μmol/L);CCK-8法检测细胞增殖活力;高内涵系统成像法检测细胞的增殖能力;激光共聚焦法检测细胞外基质沉积蛋白(ECM)Ⅳ型胶原蛋白(Col-Ⅳ)、纤维连接蛋白(FN)表达情况;Western blot法检测细胞上细胞外信号调节激酶(ERK)、p-ERK、磷酸化促分裂原激活激酶1(p-MSK1)、IκB、p-IκB、p65、p-p65的蛋白表达情况。结果CCK-8法结果显示,BBR(30、60、90μmol/L)均可抑制MCs的增殖活力;高内涵系统成像结果显示,BBR(30、60、90μmol/L)可抑制HG诱导的MCs的异常增殖;激光共聚焦实验结果显示,BBR(90μmol/L)能降低ECM上的Col-Ⅳ、FN的表达水平;Western blot实验结果显示,BBR(30、60、90μmol/L)能降低MCs上p-ERK、p-MSK1、p-IκB、p-p65蛋白的表达水平。结论BBR可能调节MCs上的ERK/NF-κB的信号通路,改善HG诱导的MCs异常增殖。

Objective To investigate the regulation effect of berberine(BBR) on abnormal proliferation of mesangial cells(MCs) induced by high glucose(HG) and its possible mechanism. Methods MCs were cultured in vitro. The experiment was divided into control group, HG model group and BBR different concentration gradient action group(30, 60, 90 μmol/L). CCK-8 method was used to detect cell viability;high content system imaging method was used to detect cell proliferation;laser confocal method was used to detect extracellular matrix(ECM) deposition protein collagen Ⅳ(Col-Ⅳ) and fibronectin(FN) expression;Western blot was used to detect extracellular signal-regulated kinase(ERK),p-ERK, Phosphorylated mitogen-activated kinase 1(p-MSK1), IκB, p-IκB, p65, p-p65 protein expression. Results The results of CCK-8 method showed that BBR(30,60,90 μmol/L) could regulate the proliferation of MCs;high-content system imaging results showed that BBR(30,60,90 μmol/L)could inhibit the abnormal proliferation of MCs induced by HG;the results of laser confocal experiments showed BBR(90 μmol/L) could reduce the expression of ECM deposition proteins Col-Ⅳ, FN;Western blot results showed that BBR(30,60,90 μmol/L) could reduce the expression of p-ERK, p-MSK1, p-IκB,p-p65 protein. Conclusion BBR may regulate the ERK/NF-κB signaling pathway of MCs and improve the abnormal proliferation of MCs induced by HG.