四神丸对脾肾阳虚泄泻大鼠胃窦组织Ghrelin-CaM-MLCK信号通路的影响

Effects of Sishen Pills on Ghrelin-Ca M-MLCK Signaling Pathway in Sinuses Ventriculi of Rats with Diarrhea of Yang Deficieney of Spleen and Kidney

目的观察四神丸对脾肾阳虚型泄泻模型大鼠胃窦组织Ghrelin-Ca M-MLCK信号通路的影响,探讨其作用机制。方法采用灌服腺嘌呤和番泻叶水煎剂制作脾肾阳虚型泄泻大鼠模型。将72只SD大鼠随机分为正常组、模型组、阳性药组和四神丸高、中、低剂量组,每组12只。各给药组给予相应药物灌胃,连续14 d,免疫组化和Western blot分别检测胃窦组织脑肠肽胃促生长素(Ghrelin)及其受体(GHSR)、钙调素(Ca M)、肌球蛋白轻链激酶(MLCK)的蛋白表达。结果模型组大鼠胃窦组织Ghrelin、GHSR、Ca M和MLCK蛋白表达均较正常组明显升高(P<0.01);各给药组上述指标蛋白表达均明显低于模型组(P<0.05,P<0.01),且以四神丸高剂量组最为明显。结论四神丸对脾肾阳虚泄泻模型大鼠的作用可能与调节Ghrelin-Ca M-MLCK信号通路指标的蛋白表达有关。

Objective To explore the effects of Sishen Pills on Ghrelin-CaM-MLCK signaling pathway with sinuses ventriculi in diarrhea rats of yang deficieney of spleen and kidney;To discuss its mechanism of action.Methods A model rats with diarrhea model of yang deficieney of spleen and kidney was established by treated with intragastric administration of adenine and Senna water decoction to replicate.72SD rats were randomly divided into normal group,model group,positive medicine group,Sishen Pills high-,medium-and low-dose groups,with12rats in each group.Each medication group was given relevant medicine for gavage,for14days.Immunohistochemistry and Western blot were used to detect the expressions of Ghrelin and GHSR,CaM,and MLCK protein in the sinuses ventriculi.Results In the model group,Ghrelin,GHSR,CaM and MLCK protein expressions of the sinuses ventriculi significantly increased(P<0.01).In each medication group,protein expression of the indexes was lower than the model group(P<0.05,P<0.01).The Sishen Pills high-dose group was particularly obvious.Conclusion The effect of Sishen Pills on diarrhea model rats of yang deficieney of spleen and kidney may be associated with regulation of index protein expression in Ghrelin-CaM-MLCK signaling pathway.