灭幽汤对幽门螺杆菌相关性胃炎脾胃湿热证模型小鼠TLRs/NF-κB65信号通路的影响

Effect of Mieyou decoction on TLRs/NF-κB65 signal pathway in mice with Helicobacter pylori associated gastritis with splenogastric hygropyrexia syndrome

目的:观察灭幽汤对幽门螺杆菌(Helicobacter pylori,H.pylori)相关性胃炎脾胃湿热证模型小鼠Toll样受体2(Toll-like receptor 2,TLR2)、TLR4、核转录因子-κB65(nuclear factorkappa B 65,NF-κB65)蛋白及mRNA,白介素(interleukin,IL)-6、IL-8表达的影响,探讨灭幽汤治疗H.pylori相关性胃炎脾胃湿热证的机制.方法:将70只BALB/c小鼠随机分为对照组、模型组、高浓度灭幽汤组(高灭组)、低浓度灭幽汤组(低灭组)、胃三联组(替硝唑+克拉霉素+枸橼酸铋钾颗粒),每组14只;采用复合因素(肥甘食物+湿热环境+H.pylori)建立BALB/c小鼠H.pylori相关性胃炎脾胃湿热证模型;造模成功后连续给药14 d后,分别采用免疫组织化学检测TLR2、TLR4、NF-κB65蛋白,qPCR检测TLR2 mRNA、TLR4mRNA、NF-κB65 mRNA的表达情况,采用ELISA检测血清IL-6、IL-8的表达.结果:模型组TLR2、TLR4、NF-κB65的蛋白及mRNA、血清IL-6、IL-8含量表达均高于对照组,差异有统计学意义;通过药物治疗后,低灭组、高灭组、胃三联组TLR2、TLR4、NF-κB65的蛋白及mRNA、血清IL-6、IL-8含量表达均低于模型组,差异有统计学意义;高灭组TLR2、TLR4、NF-κB65的蛋白及mRNA、血清IL-6、IL-8含量表达虽低于胃三联组,但差异无统计学意义.结论:灭幽汤可能通过干预TLRs/NF-κB65信号通路抑制TLR2、TLR4、NF-κB65及下游因子的表达以达到治疗H.pylori相关性胃炎脾胃湿热证的目的.

AIM： To detect the gastric expression of Toll-like receptor 2（TLR2）,TLR4,nuclear factor-kappa B 65（NF-κB65）,and serum levels of interleukin-6（IL-6） and IL-8 in mice with Helicobacter pylori（H.pylori） associated gastritis with splenogastric hygropyrexia syndrome（SGHS）,and to investigatethe mechanism underlying the therapeutic effect of Mieyou decoction in the treatment of chronicgastritis.METHODS： BALB/c mouse were randomly divided into five groups： a control group,a modelgroup,a high-concentration Mieyou decoction group,a low-concentrations Mieyou decoctiongroup,and a triple therapy group（tinidazole,clarithromycin and bismuth potassium citrategranules）,with 14 rats in each group.The BALB/c model of H.pylori-associated gastritis withSGHS using a composite method（fatness food ＋hygropyrexia environment ＋ H.pylori）.After thesuccess of modeling,continuous administration14 d.Immunohistochemistry was used to detectthe expression TLR2,TLR4 and NF-κB65 proteinsin gastric tissue; qPCR was used to detect theexpression of TLR2,TLR4 and NF-κB65 mRNAs;ELISA was used to detect the levels of IL-6 andIL-8 in serum.RESULTS： Compared with the control group,the expression of TLR2,TLR4,NF-κB65 pro-teins and mRNAs,and serum levels of IL-6and IL-8 in the model group were significantlyincreased.After therapy,the expression ofTLR2,TLR4,NF-κB65 proteins and mRNAs,and serum levels of IL-6 and IL-8 in the high-concentration Mieyou decoction group,low-concentration Mieyou decoction group,and tri-ple therapy group were significantly decreasedcompared with the model group.The expres-sion of TLR2,TLR4,NF-κB65 proteins andmRNAs,and serum levels of IL-6 and IL-8 inthe high-concentration Mieyou decoction groupwere higher than those in the triple therapygroup,but the differences were not statisticallysignificant.CONCLUSION： Mieyou decoction has a rolein the treatment of H.pylori-associated gastritis with SGHS possibly by intervening the TLRs/NF-κB65 signal pathway.