黄连素通过PI3K/Akt/mTOR信号通路抑制高糖条件下的肾脏系膜细胞增殖

Berberine inhibited the proliferation of glomerular mesangial cells under high glucose conditions though PI3 K/Akt/mTOR signal pathway

目的：探讨黄连素（berberine,BBR）对高糖条件下的大鼠肾脏系膜细胞 HBZY-1 cells （MCS）增殖的影响及其作用机制。方法将对数生长期细胞分成正常糖处理组（NG 组）,高糖组（HG组）和黄连素组（BBR组）。NG组细胞体系常规培养培养,HG 组细胞体系加入40 mmol/L 葡萄糖,BBR组细胞培养体系中加入40 mmol/L葡萄糖＋黄连素30μmol/L。各组细胞培养48 h 后,利用MTT检测各组HBZY-1细胞增殖；RT-PCR检测各组细胞p85,Akt和mTOR基因表达；West-ern blotting检测细胞中 p85、p-p85、Akt、p-Akt,mTOR 和 Collagen-Ⅳ蛋白的变化。结果 NG 组与HG组增值率分别为（25%±3%）和（75%±5%）,与 NG组增值率比较,HG组肾脏系膜细胞增殖率明显增高（P〈0.05）,p85与 Akt mRNA表达水平和蛋白水平均无明显变化（P＆gt;0.05）,但 p-p85、p-Akt,Collagen-Ⅳ蛋白表达水平和 mTOR mRNA 表达与蛋白水平均明显增加（P〈0.05）；BBR 组增值率为（42%±5%）,与 HG 组比较,BBR 组肾脏系膜细胞增值率明显降低（P〈0.05）,但 p-p85、p-Akt、Collagen-Ⅳ蛋白表达水平和 mTOR mRNA 表达与蛋白水平均明显下降（P〈0.05）,而 p85和Akt mRNA和蛋白表达水平均无统计学差异（P〈0.05）。结论黄连素能抑制高糖导致的系膜细胞增殖,其作用可能是通过抑制PI3K/Akt/mTOR信号通路激活。

Objective To explore the effect and mechanism of berberine （BBR）on the prolifera-tion of glomerular mesangial cells HBZY-1 （MCs）cultured under high glucose conditions.Methods The HBZY-1 （MCs）cells at logarithmic growth phase were randomly divided into NG,HG and BBR groups.The cells in the NG group were normally treated,and those in HG and FN groups were incuba-ted with high glucose （40 mmol/L）and high glucose （40 mmol/L）＋berberine （30μmol/L）,respec-tively.The proliferation of the MCSs was measured by MTT.The mRNA levels of p85,Akt and mTOR expression were detected by RT-PCR.The expression of p85,p-p85,Akt,p-Akt,mTOR and collagen-Ⅳ was detected by Western blotting.Results As compared with the NG group,the prolifera-tion rate of HBZY-1 cells and protein expression level of p-p85,p-Akt,mTOR and collagen-Ⅳ in HG group were significantly increased with the increases in mRNA expression levels of mTOR （P〈0.05）,but there was no significant difference in the mRNA and expression levels of p85 and Akt.As compared with the HG group,the proliferation rate of HBZY-1 cells and protein expression levels of p-p85,p-Akt,mTOR and collagen-Ⅳ in BBR group were significantly decreased with the decrease in the mRNA expression level of mTOR （P〈0.05）,but there was no significant difference in the mRNA and protein expression of p85 and Akt.The proliferation rate of HBZY-1 （MCs）in NG,HG and BBR groups was （25%±3%）,（75%±5%）and （42%±5%）,respectively.Conclusions Berberine could inhibit the proliferation of HBZY-1 cells and the effect may be achieved through attenuating PI3K/Akt/mTOR signal transduction pathway activation.