摘要
目的:观察半枝莲总黄酮(TF-SB)对高糖诱导的足细胞(MPC-5)损伤及Smad4/PKM2/HIF-1α信号通路的影响。方法:首先采用CCK8法分析TF-SB对MPC-5细胞的安全性及药效浓度。随后MPC-5分为空白组、模型组和TF-SB组,除空白组外,模型组和TF-SB组采用高糖诱导建立MPC-5细胞损伤模型,分别检测TF-SB对MPC-5细胞ATP、凋亡和ROS水平的影响,并采用酶联免疫吸附试验(ELISA)测定细胞因子(IL-1β、TNF-α、MCP-1)含量,反转录聚合酶链反应(RT-PCR)检测糖酵解基因(GLU1、PFK1和HK1)表达丰度,蛋白质印迹法(Western blot)检测各组细胞Smad4/PKM2/HIF-1α信号通路中相关蛋白的表达水平;结果:与空白组相比,模型组MPC-5细胞ATP含量、GLU1、PKF1和HK1表达丰度显著下降,凋亡和ROS水平、IL-1β、TNF-α与MCP-1的含量均显著增加(P<0.01);与模型组相比,TF-SB组ATP含量、GLU1、PKF1和HK1表达丰度显著升高,凋亡和ROS水平、IL-1β、TNF-α与MCP-1的含量均显著降低(P<0.01)。此外,与空白组相比,模型组Smad4、HIF-1α蛋白表达及细胞核中PKM2表达显著升高,细胞质中PKM2表达显著降低(P<0.01);与模型组相比,TF-SB组Smad4、HIF-1α表达及细胞核中PKM2表达显著降低,而细胞质中PKM2表达显著升高(P<0.01);结论:TF-SB促进MPC-5细胞线粒体活性诱导糖酵解,进而抑制炎症的分泌,达到治疗糖尿病肾病的作用可能与通过抑制Smad4/PKM2/HIF-1α信号通路有关。
Objective:To observe the effect of total flavonoids of Scutellaria barbata(TF-SB)on the injury of high glucose induced podocytes(MPC-5)and the influence of Smad4/PKM2/HIF-1αpathway.Methods:Firstly,CCK8 was used to analyze the safety and efficacy concentration of TF-SB on MPC-5 cells.Then,MPC-5 was then divided into the control group,model group and TF-SB group.In addition to the control group,model group and TF-SB group were induced by high glucose to establish MPC-5 cell injury model.The effects of TF-SB on ATP,apoptosis and ROS levels of MPC-5 cells were detected respectively.The contents of IL-1β,TNF-α,and MCP-1 were determined by ELISA,the expression abundance of glycolytic genes(GLU1,PFK1 and HK1)were detected by RT-PCR.Western blot method was used to detect the expression level of related proteins in Smad4/PKM2/HIF-1αpathway.Results:Compared with the blank group,ATP content,GLU1,PKF1 and HK1 expression abundance of MPC-5 cells in the model group decreased significantly,apoptosis,ROS level and IL-1β、TNF-αAnd MCP-1 significantly increased(P<0.01);Compared with model group,ATP content,GLU1,PKF1 and HK1 expression abundance,apoptosis,ROS level and IL-1βin TF-SB group were significantly increased,TNF-αThe contents of MCP-1 and MCP-1 decreased significantly(P<0.01).In addition,compared with the blank group,the model group Smad4 and HIF-1αThe protein expression and PKM2 expression in nucleus were significantly increased,while PKM2 expression in cytoplasm was significantly decreased(P<0.01);Compared with model group,TF-SB group Smad4,HIF-1αThe expression of PKM2 in the nucleus and expression of PKM2 were significantly decreased,while the expression of PKM2 in the cytoplasm was significantly increased(P<0.01).Conclusion:TF-SB promotes the mitochondrial activity of MPC-5 cells to induce glycolysis,and then inhibits the secretion of inflammation,which may play a role in treating diabetes nephropathy by inhibiting Smad4/PKM2/HIF-1αsignaling pathway.
作者
闫东
顾长瑞
张响
YAN Dong;GU Chang‑rui;ZHANG Xiang(Heilongjiang Academy of traditional Chinese Medicine,Harbin 150000,China;Heilongjiang Ruijing Diabetes Hospital,Harbin 150000,China)
出处
《海南医学院学报》
CAS
2023年第2期102-108,共7页
Journal of Hainan Medical University
基金
黑龙江省中医药科研项目(ZHY19-058)。
