疏肝和胃方对GERD大鼠食管运动功能的影响及机制研究

Study on the Effect and Mechanism of Shugan Hewei Formula on Esophageal Motility in GERD Rats

目的探讨疏肝和胃方对GERD大鼠食管运动功能的影响。方法大鼠随机分为空白组、模型组、疏肝和胃方组、雷贝拉唑组。采用半幽门结扎贲门肌切开术建立动物模型。HE染色观察食管组织,RT-qPCR、Western blot和免疫组化法检测PAR-2、IP3R、RyR、MLCK、CaM mRNA及蛋白表达,离体肌条组织实验探讨PAR-2激动剂对肌条的影响。结果与模型组比较,疏肝和胃方组与雷贝拉唑组PAR-2 mRNA和蛋白表达降低(P<0.05),CaM、MLCK的mRNA和蛋白表达升高(P<0.05),IP3R的蛋白表达升高(P<0.05),同时疏肝和胃方组IP3R mRNA表达升高(P<0.05);离体肌条组织实验结果显示,疏肝和胃方组和雷贝拉唑组肌条肌张力变化率上升(P<0.05),PAR-2激动剂对两组肌条的影响下降(P<0.05)。结论疏肝和胃方治疗GERD的机制可能和调控PAR-2信号通路有关。

Objective Study on the effect of Shugan Hewei formula on esophageal motility function in GERD rats.Methods Rats were randomly divided into blank group,model group,Shugan Hewei formula group,and rabeprazole group.The animal model was established using semi pyloric ligation and cardiac myotomy.HE staining was used to observe esophageal tissue,and RT-qPCR,Western blot,and immunohistochemistry were used to detect the mRNA and protein expression of PAR-2,IP3R,RyR,MLCK,CaM.In vitro muscle strip tissue experiments were conducted to investigate the effect of PAR-2 agonists on muscle strips.Results Compared with the model group,the expression of PAR-2 mRNA and protein was reduced(P<0.05),while the expression of CaM and MLCK mRNA and protein was increased(P<0.05),and the expression of IP3R protein was increased(P<0.05)in the Shugan Hewei formula group and the rabeprazole group.At the same time,the expression of IP3R mRNA was increased(P<0.05)in the Shugan Hewei group;The results of in vitro muscle strip tissue experiments showed that the change rate of muscle strip tension in the Shugan Hewei formula group and rabeprazole group increased(P<0.05),while the effect of PAR-2 agonist on the two groups of muscle strips decreased(P<0.05).Conclusion The mechanism of Shugan Hewei Formula in treating GERD may be related to the regulation of PAR-2 signaling pathway.