糖通饮含药血清对高糖环境下HBZY-1细胞增殖的影响及其作用机制

Effect of tangtongyin drug-containing serum on proliferation of HBZY-1 cells in high glucose environment and its mechanism

目的分析糖通饮含药血清对高糖环境下大鼠肾小球系膜细胞(HBZY-1)增殖的影响及可能作用机制。方法将30只SPF级雄性SD大鼠随机均分为空白血清组和糖通饮含药血清组,糖通饮含药血清组大鼠予以糖通饮灌胃,空白血清组大鼠予以等体积的双蒸水灌胃,灌胃7 d后股动脉取血;常规培养第3~15代HBZY-1细胞分为正常糖+空白血清组、高糖+空白血清组以及高糖+糖通饮含药血清组,按分组给予相应大鼠血清干预;细胞培养结束后,CCK-8法检测各组细胞增殖情况,实时荧光定量PCR法检测各组细胞转化生长因子-β1(TGF-β1)、Smad 2、Smad 3 mRNA表达,Western blot法检测各组细胞TGF-β1、p-Smad2/Smad2、p-Smad3/Smad3蛋白表达。结果与正常糖+空白血清组相比,高糖+空白血清组HBZY-1细胞明显增殖,细胞中TGF-β1、Smad2、Smad3 mRNA表达增加,TGF-β1、p-Smad2/Smad2、p-Smad3/Smad3蛋白表达也增加,差异均具有统计学意义(P<0.05);与高糖+空白血清组相比,高糖+糖通饮含药血清组HBZY-1细胞增殖被抑制,细胞中的TGF-β1、Smad2、Smad3 mRNA表达降低,TGF-β1、p-Smad2/Smad2、p-Smad3/Smad3蛋白表达也降低,差异均具有统计学意义(P<0.05)。结论糖通饮可以抑制高糖状态下HBZY-1细胞的增殖,其作用机制可能与抑制TGF-β1/Smad信号通路的过度激活有关。

Objective To analyze the effect of tangtongyin drug-containing serum serum on the proliferation of rat glomerular mesangial(HBZY-1)cells in high glucose environment and its possible mechanism.Methods Thirty SPF male SD rats were randomly divided into blank serum group and tangtongyin drug-containing serum group.Rats in tangtongyin drug-containing group were fed with tangtongyin by intragastric administration,and rats in blank serum group were given equal volume double distilled water.Blood was collected from femoral artery after continuous intragastric administration for 7 days.HBZY-1 cells cultured from 3 rd to 15 th generation were divided into normal glucose+blank serum group,high glucose+blank serum group and high glucose+tangtongyin drug-containing serum group.At the end of cell culture,the cell proliferation of each group was detected by CCK-8 method,the expression of TGF-β1,Smad 2,and Smad 3 mRNA was detected by real-time fluorescence quantitative PCR,and the expression of TGF-β1,p-Smad2/Smad2,and p-Smad3/Smad3 protein was detected by Western blot method.Results Compared with the normal glucose+blank serum group,cells in the high glucose+blank serum group proliferated significantly,and the expression of TGF-β1,Smad 2,and Smad 3 mRNA and the expression of TGF-β1,p-Smad2/Smad2,and p-Smad3/Smad3 protein were also increased(P<0.05).Compared with the high glucose+blank serum group,the cell proliferation was inhibited,and the expression of TTGF-β1,Smad 2,and Smad 3 mRNA and the protein expression of TGF-β1,p-Smad2/Smad2,and p-Smad3/Smad3 were decreased in the high glucose+tangtongyin group(P<0.05).Conclusion Tangtongyin can inhibit the proliferation of hbzy-1 cells in high glucose state,and its mechanism may be related to the inhibition of TGF-β1/Smad signaling pathway.