大黄提取物调控JAK2/STAT6通路对脂多糖诱导巨噬细胞极化的影响  

Effect of Rhubarb Extract on Lipopolysaccharide Induced Macrophage Polarization by Regulating the JAK2/STAT6 Pathway

作　　者：周甜 蒋婷 华丰 刘产明[1] 朱月琴 张晓鸣[1] 戴亦娴 ZHOU Tian;JIANG Ting;HUA Feng;LIU Chanming;ZHU Yueqin;ZHANG Xiaoming;DAI Yixian(Department of Pharmacy,Changzhou Hospital of Traditional Chinese Medicine,Nanjing University of Chinese Medicine,Changzhou 213000,China)

机构地区：[1]南京中医药大学附属常州市中医医院药学部,江苏常州213000

出　　处：《药物生物技术》2025年第1期1-6,共6页Pharmaceutical Biotechnology

基　　金：国家中药炮制技术传承基地建设项目([2022]45号);江苏省中医药管理局项目(No.202232);常州市科技计划项目(No.2021157)。

摘　　要：探讨大黄提取物(RE)调控酪氨酸激酶2(JAK2)/信号转导和转录激活因子6(STAT6)通路对脂多糖(LPS)诱导巨噬细胞极化的影响。取融合度达到70%的RAW264.7细胞,分为对照组、LPS组、RE-L组、RE-M组、RE-H组、RE-H+AG490(JAK2阻断剂)组。采用CCK-8法检测RAW264.7细胞活力,免疫荧光检测细胞中CD86、CD206表达,qRT-PCR检测细胞中诱导型一氧化氮合酶(iNOS)mRNA、精氨酸-1(Arg-1)mRNA表达,ELISA法检测细胞上清中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6、一氧化氮(NO)、IL-10水平,Western blot检测细胞中p-JAK2、p-STAT6蛋白表达水平。与对照组相比,LPS组RAW264.7细胞活力、CD206表达的相对荧光强度、Arg-1 mRNA表达、IL-10水平及p-JAK2、p-STAT6蛋白表达降低,CD86表达的相对荧光强度、iNOS mRNA表达、TNF-α、IL-6、NO水平升高(P<0.05);与LPS组相比,RE-L组、RE-M组、RE-H组RAW264.7细胞活力、CD206表达的相对荧光强度、Arg-1 mRNA表达、IL-10水平及p-JAK2、p-STAT6蛋白表达升高,CD86表达的相对荧光强度、iNOS mRNA表达、TNF-α、IL-6、NO水平降低(P<0.05);与RE-H组相比,RE-H+AG490组RAW264.7细胞活力、CD206表达的相对荧光强度、Arg-1 mRNA表达、IL-10水平及p-JAK2、p-STAT6蛋白表达降低,CD86表达的相对荧光强度、iNOS mRNA表达、TNF-α、IL-6、NO水平升高(P<0.05)。研究证实,RE促进LPS诱导的RAW 264.7细胞由M1型向M2型转化的机制可能与激活JAK2/STAT6通路有关。To investigate the effect of rhubarb extract(RE)on lipopolysaccharide(LPS)induced macrophage polarization by regulating the tyrosine kinase 2(JAK2)/signal transducer and activator of transcription 6(STAT6)pathway,RAW264.7 cells with a fusion degree of 70%were selected and separated into control group,LPS group,RE-L group,RE-M group,RE-H group,and RE-H+AG490(JAK2 blocker)group.CCK-8 method was applied to detect RAW264.7 cell viability.Immunofluorescence was applied to detect the expression of CD86 and CD206 in cells.QRT-PCR was applied to detect the expression of inducible nitric oxide synthase(iNOS)mRNA and arginine-1(Arg-1)mRNA in cells.ELISA method was applied to detect the levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),nitric oxide(NO),and IL-10 in the cell supernatant.Western blot was applied to detect the expression of p-JAK2 and p-STAT6 proteins in cells.Compared with the control group,the RAW264.7 cell viability,relative fluorescence intensity of CD206 expression,the mRNA expression of Arg-1,level of IL-10,and the protein expression of p-JAK2 and p-STAT6 were reduced in the LPS group;the relative fluorescence intensity of CD86 expression,the mRNA expression of iNOS,levels of TNF-α,IL-6,and NO were increased(P<0.05).Compared with the LPS group,the RAW264.7 cell viability,relative fluorescence intensity of CD206 expression,the mRNA expression of Arg-1,level of IL-10,and the protein expression of p-JAK2 and p-STAT6 were increased in RE-L,RE-M,and RE-H groups;the relative fluorescence intensity of CD86 expression,the mRNA expression of iNOS,levels of TNF-α,IL-6,and NO were decreased(P<0.05).Compared with the RE-H group,the RAW264.7 cell viability,relative fluorescence intensity of CD206 expression,the mRNA expression of Arg-1,level of IL-10,and the protein expression of p-JAK2 and p-STAT6 were reduced in the RE-H+AG490 group;the relative fluorescence intensity of CD86 expression,the mRNA expression of iNOS,levels of TNF-α,IL-6,and NO were increased(P<0.05).The mechanism by which RE promotes

关 键 词：大黄提取物 酪氨酸激酶2/信号转导和转录激活因子6通路 脂多糖 巨噬细胞 细胞极化 炎症 

分 类 号：R285.5[医药卫生—中药学]