灭幽汤对幽门螺杆菌相关性胃炎脾胃湿热证小鼠NF-κB65、HSP70的影响

Effect of Mieyou Soup on NF-κB65 and HSP70 in the H.Pylori-associated gastritis mice of Pi-Wei hygropyrexia syndrome(PWHS)

[目的]研究灭幽汤对幽门螺杆菌(helicobacter pylori,Hp)相关性胃炎脾胃湿热证小鼠核因子-κB65(NF-κB65)及热休克蛋白70(HSP70)的相关性,探讨Hp相关性胃炎脾胃湿热证的发病机制及灭幽汤治疗慢性胃炎的作用机制。[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+幽门螺杆菌)建立BALB/c小鼠Hp相关性胃炎脾胃湿热证模型;造模成功并连续给药14d后,分别采用免疫组织化学、Western Blot检测NF-κB65蛋白、HSP70蛋白,qRT-PCR检测NF-κB65mRNA、HSP70mRNA的表达情况。[结果]HSP70蛋白及其mRNA的表达:与对照组比较,模型组HSP70蛋白及其mRNA表达均显著增加(P<0.01);与模型组比较,高灭组、低灭组、胃三联组小鼠HSP70蛋白及其mRNA表达均增加(P<0.01);高灭组HSP70及其mRNA表达高于低灭组(P<0.05);胃三联组、高灭组HSP70蛋白及其mRNA表达均增加,但2组间比较差异无统计学意义。NF-κB65蛋白及其mRNA的表达:与对照组比较,模型组NF-κB65蛋白及其mRNA表达增加(P<0.01);与模型组比较,高灭组、胃三联组小鼠NF-κB65蛋白及其mRNA表达均明显降低(P<0.01);高灭组NF-κB65蛋白及其mRNA表达低于低灭组(P<0.05);高灭组、胃三联组NF-κB65蛋白及其mRNA均降低,但2组间比较差异无统计学意义。[结论]灭幽汤可能通过上调HSP70、下调NF-κB65表达而发挥治疗Hp相关性胃炎脾胃湿热证的作用。

[Objective]To study the expressions of NF-kappaB65 （NF-κB65）and heat shock protein70 （HSP 70）in the H. Pylori-sssociated gastritis mice of Pi-Wei hygropyrexia syndrome（PWHS）, and to in- vestigate the mechanism of Mieyou Soup in the treatment of chronic gastritis. [Methods]BALB/c mice were randomly divided into five groups：control group, model group, low concentration of Mieyou soup group, high concentration of Mieyou soup group,gastric triad group（Tinidazole, Clarithromycin and Bismuth po- tassium citrate granules）, with 14 rats in each group. Pi-Wei hygropyrexia syndrome mouse model of H. Pylori-associated gastritis was established by composite factor（fatness foodq-hygropyrexia environment ＋ Helicobacter pylori） After the success of modeling,corresponding treatment was given to mice in each group for 14 days. Immunohistochemistry was used to detect the expression of NF-κB65 protein,western blot to detect the expression of HSP70 protein,and qRT-PCR to detect the expression of NF-κB65 mRNA and HSP70 mRNA. rResultsκCompared with control group,the expression of HSP70 protein and HSP70 mRNA in BALB/c mice model group was significantly increased （P〈0. 01） Compared with the model group,the HSP70 protein and HSP70 mRNA in high and low concentrations of Mieyou soup group as well as the gastric triad group were increased, and the differences were statistically significant（P〈0. 01） The expression of HSP70 protein and HSP70 mRNA in high-concentration Mieyou soup group was higher than low-concentrations group（P〈0.05） The expression of HSP70 protein and HSP70 mRNA in gastric triad group and high-concentration Mieyou soup group were increased, but the difference was insignificant（P〉 0.05）. Compared with control group,the expression of NF-κB65 protein and NF-κB65 mRNA in the model group was significantly increased（P〈0.01） Compared with the model group, the NF-~B65 protein and NF- κB65 mRNA in high-concentration Mieyou soup group and gastric triad group were decreased, and the differences were statistically significant（P〈0.01） The expression of NF-κB65 protein and NF-κB65 mRNA in high-concentration Mieyou soup was lower than those of low-concentration Mieyou soup group （P 0.05） The expression of NF-κB65 protein and NF-κB65 mRNA in gastric triad group and high-concentra- tion Mieyou soup were decreased, but the difference was not statistically significant （P〉 0.05）. EConelu- sion^Mieyou Soup may play a role in the treatment of H. Pylori-associated gastritis mice of Pi-Wei hygro- pyrexia syndrome by raising the expression of HSP70,reducing the expression of NF-κB65 protein expres- sion.