半夏泻心汤调控巨噬细胞分泌HSP70抗Hp性胃炎作用机制研究

Effect of Banxiaxiexin Decoction on H.pylori gastritis by regulating the secretion of HSP70

[目的]探讨半夏泻心汤体外杀幽门螺杆菌作用及调节巨噬细胞分泌应激性细胞因子抗胃炎的作用机制。[方法]固体稀释法测定体外半夏泻心汤对幽门螺杆菌的最低杀菌浓度(MBC)。用荧光定量PCR和WesternBlot的方法,检测含药血清作用原代培养的小鼠巨噬细胞后,巨噬细胞内HSP70细胞因子表达变化情况。[结果]体外杀菌实验测得半夏泻心汤对幽门螺杆菌的最低杀菌浓度为45mg/ml。荧光定量PCR结果表明,5%含药血清作用3h、6h时,均能抑制巨噬细胞分泌HSP70(P<0.01);而10%含药血清作用3h、6h均不能抑制巨噬细胞分泌HSP70(P>0.05)。Western-Blot结果表明,5%含药血清作用巨噬细胞3h、6h时,巨噬细胞内HSP70表达均下调。而10%含药血清作用巨噬细胞3h、6h时,巨噬细胞内HSP70表达均没有改变。[结论]半夏泻心汤在体外有杀幽门螺杆菌作用,其最低杀菌浓度为45mg/ml。其抗胃炎作用机制可能是如下两方面:一方面半夏泻心汤杀死胃黏膜下层的幽门螺杆菌,另一方面,半夏泻心汤调节巨噬细胞的活性,抑制巨噬细胞分泌HSP70应激因子,减轻胃黏膜上皮细胞的炎性损伤。此研究结果将初步阐明了半夏泻心汤的抗胃炎机制。

[Objective]To investigate the effect of Banxiaxiexin Decoction（BD）on Helicobacter pylori in vitro and its mechanism of anti-gastritis by regulating macrophage secretion stress cytokine.[Methods]The minimum bactericidal concentration（MBC）of BD for killing Helicobacter pylori in vitro was determined by solid dilution method.Fluorescence quantitative PCR and Western Blot methods were applied to test HSP70 cytokine expression in primary cultured mouse macrophages affected by drug-containing serum.[Results]Bactericidal experiment in vitro measured that MBC of BD was 45mg/ml.Fluorescence quantitative PCR results showed that action of 5% drug-containing serum for 3hor 6hwas able to inhibit HSP70 secretion of macrophage（P〈0.01）,while 10% drug-containing serum for for 3hor 6hdid not（P〈0.05）Western Blot results showed that after 5% drug-containing serum acted on macrophages for 3hor 6h,HSP70 expression in macrophages were down-regulated,while with action by 10% drug-containing serum,HSP70 expression in macrophages did not change.[Conclusion]The BD had the effect of killing Helicobacter pylori in vitro with MBC 45mg/ml.The mechanism of its anti-gastritis action may be related to the following two aspects：on the one hand,BD killed the Helicobacter pylori of gastric mucosa,on the other hand,it regulated the activity of macrophages,inhibited the HSP70 stress cytokines secreted by macrophages and reduced the inflammatory injury of gastric mucosal epithelial cells.