摘要
目的 研究黄芩汤(HQT)对结肠上皮细胞炎症模型上皮间质转化进程的影响,并探讨其药效及相关作用机制。方法 使用脂多糖(lipopolysaccharide,LPS)诱导结肠上皮细胞IEC-6制备炎症模型并分别给予高、中、低剂量(500、250、125μg·mL^(-1))黄芩汤干预后,ELISA法检测肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)含量,Western-Blot法检测海马无翅基因3A(recombinant wingless type MMTV integration site family member 3A,WNT3A)、β-连环蛋白(β-catenin)、E-钙黏素(E-cadherin)、糖原合成酶激酶3β(glycogen synthase kinase-3β,GSK3β)、前梯度蛋白2(anterior gradient-2,AGR2)、紧密连接蛋白-1(claudin-1,Cldn-1)的蛋白表达,qRT-PCR法检测WNT3A、β-catenin、E-cadherin、GSK3β、AGR2、Cldn-1的mRNA表达。结果 与模型组相比,黄芩汤组细胞内炎症因子IL-1β、TNF-α含量均明显降低(P<0.01)且存在一定剂量依赖性;黄芩汤组细胞内E-cadherin、GSK-3β蛋白和mRNA表达水平均明显升高(P<0.01),β-catenin、AGR2蛋白和mRNA表达水平均明显降低(P<0.01),以高剂量黄芩汤组效果最为显著。结论 黄芩汤可有效遏制炎症环境下结肠上皮细胞中异常的上皮间质转化进程,这一作用机制可能与抑制经典WNT通路中WNT3A、β-catenin、AGR2、Cldn-1表达,同时促进E-cadherin、GSK3β表达有关。
Objective To investigate the effect of Huangqin Decoction(HQT)on epithelialmesenchymal transition of colonic epithelial cells inflammatory model.Methods Lipopolysaccharide(LPS)was used to induce IEC-6 cells to construct colonic epithelial cells inflammatory model.After administration with HQT(500,250,125μg·mL^(-1)),Tumor necrosis factor-α(TNF-α),Interleukin-1β(IL-1β)content were detected by ELISA;Western blotting was used to detect recombinant wingless type MMTV integration site family member 3A(WNT3A),β-catenin,E-cadherin,glycogen synthase kinase3β(GSK3β),Anterior gradient-2(ACR2),Claudin-1(Cldn-1)protein expressions;qRT-PCR was used to detect WNT3A,β-catenin,E-cadherin,AGR2,Cldn-1 mRNA expressions.ResultssCompared with model group,the levels of intracellular inflammatory factors IL-1βand TNF-αwere significantly reduced in the cells of HQT group(P<O.01),and there was a certain dose-dependence;The intracellular expression levels of E-cadherin,GSK-3βprotein and mRNA in the HQT group.were significantly higher(P<0.01),and the expression levels ofβ-catenin,AGR2 protein and mRNA were significantly lower(P<0.01),the most significant effect was seen in the high-dose HQT group.Conclusionn HQT can effectively inhibit the excessive epithelial mesenchymal transition of colonic epithelial cells inflammatory model,and its mechanism maybe related to reducing the expressions of WNT3A,β-catenin,AGR2 and Cldn-1 in the WNT pathway and promoting the expression of E-cadherin and GSK3β.
作者
马旭冉
姜言红
杨伟鹏
王淑慧
MA Xuran;JIANG Yanhong;YANG Weipeng;WANG Shuhui(Institute of Pharmacy,Shandong University of Traditional Chinese Medicine,Ji'nan 250355;The Second Clinical Medical Institute,Shandong University of Traditional Chinese Medicine;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences;State Key Laboratory of Natural and Biomimetic Drugs,School of Pharmaceutical Sciences,Peking University)
出处
《现代中医临床》
2025年第4期110-116,共7页
Modern Chinese Clinical Medicine
基金
国家自然科学基金青年项目(No.82304690)
国家自然科学基金面上项目(No.82074328)。
关键词
黄芩汤
肠道上皮屏障
炎症
上皮间质转化
作用机制
Huangqin Decoction
intestinal epithelial barrier
inflammation
epithelial mesenchymal transition
mechanism of action
