黄芩素介导PI3K/Akt信号通路对高糖诱导HK-2细胞氧化应激和炎症反应的影响研究  

Effects of baicalein-mediated PI3K/Akt signaling pathway on high glucose-induced oxidative stress and inflammatory response in HK-2 cells

作　　者：杨柳[1] 张欣雅 王静[1] 王怡 程汝阳 李晨响 李爽[1] 赵世超 李建民[1] YANG Liu;ZHANG Xinya;WANG Jing;WANG Yi;CHENG Ruyang;LI Chenxiang;LI Shuang;ZHAO Shichao;LI Jianmin(Heilongjiang University of Chinese Medicine, Harbin 150040, Heilongjiang, China)

机构地区：[1]黑龙江中医药大学,黑龙江哈尔滨150040

出　　处：《现代中西医结合杂志》2022年第4期455-458,463,共5页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：国家自然科学基金面上项目(81274035);黑龙江中医药大学校科研基金资助项目(201820);黑龙江省卫健委科研项目(202102040369);黑龙江省大学生创新创业训练计划项目(201910228058);黑龙江中医药大学大学生科技创新项目(2021-9)。

摘　　要：目的探讨黄芩素通过磷脂酰肌醇3-激酶/蛋白激酶B(BPI3K/Akt)通路对高糖诱导的人近端肾小管上皮细胞(HK-2)氧化应激和炎症反应的调控作用。方法实验分为5组:正常组HK-2细胞只加入DMEM低糖培养基培养48 h;高糖模型组HK-2细胞在DMEM低糖培养基中加入4×10^(-2)mol/L浓度高糖培养48 h;黄芩素低、中、高剂量组HK-2细胞先分别加入1×10^(-7)mol/L、1×10^(-6)mol/L、1×10^(-5)mol/L黄芩素培养24 h后,再加入4×10^(-2)mol/L浓度高糖继续培养24 h。采用四甲基噻唑蓝(MTT)法检测各组细胞增殖率,按照试剂盒说明检测各组细胞中活性氧(ROS)、丙二醛(MDA)含量,采用逆转录-聚合酶链反应(RT-PCR)检测各组细胞中肿瘤坏死因子-α(TNF-α)mRNA表达量,采用Western blot法检测各组细胞中磷酸化磷酸肌醇3-激酶(p-PI3K)、磷酸化蛋白激酶B(p-Akt)、TNF-α蛋白表达量。结果与正常组比较,高糖模型组细胞存活率明显降低(P<0.05),细胞中ROS、MDA含量及TNF-αmRNA表达量和p-PI3K、p-Akt、TNF-α蛋白表达量均明显升高(P均<0.05);与高糖模型组比较,黄芩素各组细胞存活率均明显升高(P均<0.05),细胞中ROS、MDA含量及TNF-αmRNA表达量和p-PI3K、p-Akt、TNF-α蛋白表达量均明显降低(P均<0.05)。结论黄芩素可能通过抑制高糖诱导的氧化应激反应及PI3K/Akt通路活化来减少炎症因子产生,从而发挥保护HK-2细胞作用。Objective It is to investigate the regulation effect of baicalein on oxidative stress and inflammatory response in human proximal renal tubular epithelial cells(HK-2)induced by high glucose though phosphatidylinositol 3-kinase/protein kinase B(BPI3K/Akt)pathway.Methods The experiment was divided into 5 groups:the HK-2 cells in the normal group were cultured with DMEM low-glucose medium for 48 hours;the HK-2 cells in the high-glucose model group were cultured in DMEM low-glucose medium added with 4×10^(-2)mol/L high glucose for 48 hours;the HK-2 cells in baicalein low,medium and high dose groups were firstly cultured with 1×10^(-7)mol/L,1×10^(-6)mol/L,1×10^(-5)mol/L baicalein for 24 hours,and then added with 4×10^(-2)mol/L high glucose continued to culture for another 24 h.The cell proliferation rate in each group was detected by tetramethylthiazole blue(MTT)assay,the contents of reactive oxygen species(ROS)and malondialdehyde(MDA)in each group were detected according to kit instructions,the mRNA expression of tumor necrosis factor-α(TNF-α)in the cells of each group was detected by reverse transcription-polymerase chain reaction(RT-PCR),and the phosphorylated phosphoinositide 3-kinase(phosphoinositide 3-kinase)(p-PI3K),phosphorylated protein kinase B(p-Akt),TNF-αprotein expression in the cells of each group was detected by Western blot.Results Compared with the normal group,the cell survival rate of the high-glucose model group was significantly reduced(P<0.05),and the contents of ROS,MDA,TNF-α,p-PI3K/PI3K and p-AKT/AKT protein and mRNA in the cells were all increased(all P<0.05).Compared with the high-glucose model group,the cell survival rate was significantly increased in each baicalein group,and the contents of ROS,MDA,TNF-α,p-PI3K/PI3K and p-AKT/AKT protein and mRNA in the cells were significantly reduced(all P<0.05).Conclusion Baicalein may reduce the production of inflammatory factors by inhibiting high glucose-induced oxidative stress and activation of PI3K/Akt pathway,thereby protecting HK-2 cel

关 键 词：黄芩素 人近端肾小管上皮细胞 磷脂酰肌醇3-激酶/蛋白激酶B信号通路 氧化应激 炎症反应 

分 类 号：R-33[医药卫生]