祛风通络配伍对高糖诱导人肾小管上皮细胞活力及炎症反应的影响  

Effects of Qufeng Tongluo fomula of human renal tubular epithelial cells on cell viability and inflammation induced by high glucose

作　　者：苏冠旬 李丽娜 赵进喜[2] 陈宇 薛泰骑[1,2] 魏金艳 王淑艳 王世东 SU Guanxun;LI Lina;ZHAO Jinxi;CHEN Yu;XUE Taiqi;WEI Jinyan;WANG Shuyan;WANG Shidong(The First Clinical Medical Collage of Beijing University of Chinese Medicine,Beijing 100700,China)

机构地区：[1]北京中医药大学第一临床医学院,100700 [2]北京中医药大学东直门医院肾病内分泌科 [3]北京中医药大学中医学院

出　　处：《环球中医药》2021年第12期2121-2126,共6页Global Traditional Chinese Medicine

基　　金：北京市自然科学基金面上项目(7182096);北京市通州区卫生发展科研专项(TZ2016HZPP)。

摘　　要：目的观察祛风通络配伍对高糖诱导人肾小管上皮细胞(human renal tubular epithelial cell,HKC)活力及炎症反应的影响。方法体外培养HKC并采用高糖DMEM培养基诱导其增殖,设立正常组、模型组、抑制剂组、祛风通络组,CCK-8法测定各组细胞的活力;酶联免疫吸附测定法检测白细胞介素1β(interleukin 1β,IL-1β)、白细胞介素6(interleukin 6,IL-6)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)的含量;逆转录—聚合酶链式反应(RT-PCR)法测定IL-1β、IL-6、TNF-α、细胞间黏附分子-1(intercellular adhesion molecule 1,ICAM-1)、单核细胞趋化蛋白-1(monocyte chemoattractant protein 1,MCP-1)、p38丝裂原活化蛋白激酶(p38 mitogen activated protein kinase,p38 MAPK)的mRNA的表达;蛋白免疫印迹法检测p38 MAPK、ICAM-1、MCP-1的蛋白表达水平。结果与正常组比较,模型组细胞活力显著下降(P<0.05),IL-1β、IL-6、TNF-α的含量增加(P<0.05),IL-1β、IL-6、TNF-α、ICAM-1、MCP-1、p38 MAPK的mRNA表达增加(P<0.05),p38 MAPK、ICAM-1、MCP-1蛋白的表达增加(P<0.05);与模型组比较,抑制剂组和祛风通络组细胞活力显著增高(P<0.05),IL-1β、IL-6、TNF-α的含量降低(P<0.05),IL-1β、IL-6、TNF-α、ICAM-1、MCP-1、p38 MAPK mRNA的表达降低(P<0.05),MCP-1、p38 MAPK、ICAM-1蛋白的表达减少(P<0.05)。结论祛风通络配伍可以明显提高高糖诱导HKC的细胞活力,并可能通过作用于p38 MAPK信号通路减少IL-1β、IL-6、TNF-α、MCP-1、ICAM-1的表达,进而减轻炎症反应。Objective To observe the effects of Qufeng Tongluo fomula on human renal tubular epithelial cell(HKC)viability and inflammation induced by high glucose.Methods In vitro culture of renal tubular epithelial cells and the use of high-glucose DMEM media to induce their proliferation, the normal group, model group, inhibitor group, andQufeng Tongluogroup were established. The CCK-8 method was used to determine the viability of human renal tubular epithelial cells. The levels of interleukin 1β (IL-1β), interleukin 6(IL-6) and tumor necrosis factor α(TNF-α) were determined by enzymy-linked immunosorbent assay(Elisa). The mRNA expressions of IL-1β, IL-6, TNF-α, intercellular adhesion molecule1(ICAM-1), monocyte chemoattractant protein 1(MCP-1) and p38 mitogen activated protein kinase(p38 MAPK) were detected by reverse transcription polymerase chain reaction(RT-PCR). The expression levels of p38 MAPK, ICAM-1 and MCP-1 were detected by western blot.Results Compared with the normal group, the cell viability of the model group decreased significantly(P< 0. 05), the levels of IL-1β, IL-6 and TNF-α increased(P< 0. 05), the mRNA expressions of IL-1β, IL-6, TNF-α, ICAM-1, MCP-1 and p38 MAPK increased(P< 0. 05) and the protein expressions of p38 MAPK, ICAM-1 and MCP-1 were increased(P< 0. 05). Compared with the model group, the cell viability of the inhibitor group and the Qufeng Tongluogroup significantly increased(P< 0. 05), and the levels of IL-1β, IL-6, TNF-α, the mRNA expressions of IL-1β, TNF-α, ICAM-1, MCP-1, p38 MAPK, and the protein expressions of MCP-1,p38 MAPK and ICAM-1 protein all decreased(P< 0. 05) in both groups.Conclusions Qufeng Tongluo fomula can obviously increase the viability of renal tubular epithelial cells induced by high glucose, and may reduce the inflammatory reaction by decreasing the expressions of IL-1β, IL-6, TNF-α, MCP-1,ICAM-1 through p38 MAPK signal pathway.

关 键 词：祛风通络配伍 人肾小管上皮细胞 糖尿病肾病 从风论治 P38丝裂原活化蛋白激酶 

分 类 号：R285.5[医药卫生—中药学]