电针对脾虚湿蕴型溃疡性结肠炎大鼠水液代谢及肠黏膜通透性的调节机制

Regulatory Mechanism of Regulating Water Metabolism and Intestinal Mucosal Permeability on Electric Acupuncture of Spleen Deficiency with Dampness Retention-ulcerative Colitis Rats

观察电针“足三里”“脾俞”对脾虚湿蕴型溃疡性结肠炎(SDDR-UC)大鼠水液代谢及肠黏膜通透性的影响,并探讨其作用机制。60只SD大鼠随机分为4组:正常组、模型组、电针组和非经非穴组。正常组正常饲养,其余3组采用病证结合法复制SDDR-UC模型。电针组选取“足三里”“脾俞”穴,非经非穴组选取非经非穴点,干预过程20 min,持续14 d。观察大鼠的一般情况,测量体重、抓力、结肠长度变化、结肠黏膜损伤指数(CMDI)评分。苏木精-伊红染色(hematoxylin-eosin staining,HE)法观察结肠组织病理形态;免疫印迹(western blot,WB)法检测丝裂原活化蛋白激酶(p38MAPK)、细胞外信号调节激酶(ERK MAPK)磷酸化情况及水通道蛋白4(aquaporin protein 4,AQP4)表达水平;免疫组化(Immunohistochemistry,IHC)法检测紧密粘连蛋白1(ZO-1)、闭合蛋白(Occludin)表达水平;实时荧光定量(quantitative real-time PCR,RT-qPCR)法检测AQP4 mRNA表达量。结果表明:与正常组相比,模型组、非经非穴组大鼠体重、抓力、结肠长度下降、CMDI评分升高(P<0.01);结肠组织出现炎性细胞浸润、隐窝脓肿等病理形态表现,AQP4、ZO-1、Occludin表达降低(P<0.01),p38MAPK、ERK MAPK磷酸化程度升高(P<0.05);相较模型组,电针组大鼠体重、抓力、结肠长度提升、CMDI评分下降(P<0.01),结肠组织炎性浸润减少、隐窝边缘相对规则,AQP4、ZO-1、Occludin表达水平显著升高(P<0.01),p38MAPK、ERK MAPK蛋白磷酸化程度显著降低(P<0.05)。电针“足三里”“脾俞”可能通过抑制p38MAPK、ERK MAPK磷酸化程度,上调AQP4、ZO-1、Occludin的表达,对SDDR-UC大鼠的水液代谢及肠黏膜通透性发挥改善作用。

To observe the effects of electroacupuncture at“ST36”and“BL20”on water metabolism and intestinal mucosal permeability in colon tissue of rats with spleen deficiency and dampness-containing type ulcerative colitis(SDDR-UC)and to investigate its mechanism of action.60 SD rats were randomly divided into 4 groups:normal group,model group,non-meridian non-acupuncture group and non-meridian non-acupuncture group,electroacupuncture group and non-meridian and non-acupoint group.The normal group was fed normally,and the other three groups were replicated the SDDR-UC model using the method of combining disease and evidence.In the electroacupuncture group,the“ST36”and“BL20”were selected,and in the non-meridian and non-acupoint group,the non-meridian and non-acupoint points were selected,and the intervention process lasted 20 min for 14 d.The general conditions of the rats were observed,and the body weights,grasping strengths,changes in the lengths of the colon,and the scores of the colonic mucous membrane damage index(CMDI)scores were measured.Hematoxylin-eosin staining(HE)method was used to observe the histopathological morphology of colon.Western blot(WB)method was used to detect the phosphorylation of mitogen-activated protein kinase(p38MAPK),extracellular signal-regulated kinase(ERK MAPK),and aquaporin protein 4(AQP4),immunohistochemistry(IHC)for ZO-1 and Occludin.Quantitative real-time PCR(RT-qPCR)for AQP4.Quantitative real-time PCR(RT-qPCR)method was used to detect AQP4 mRNA expression.The results show that compared with the normal group,rats in the model group and the non-transthoracic non-cavity group show decreased body weight,grasping power,colon length and increased CMDI score(P<0.01).Colon tissues show pathological morphological manifestations such as inflammatory cell infiltration and crypt abscess,with decreased expression of AQP4,ZO-1 and Occludin(P<0.01),and phosphorylation of p38MAPK,ERK MAPK elevated(P<0.05).Compared with the model group,rats in the electroacupuncture group show elevated body weight,grip strength,colon length,decreased CMDI score(P<0.01),reduced inflammatory infiltration of colon tissue,relatively regular crypt margins,significantly higher expression levels of AQP4,ZO-1,Occludin(P<0.01),and phosphorylation of p38MAPK,ERK MAPK proteins significantly decrease(P<0.05).Electroacupuncture of“ST36”and“BL20”may play an important role in improving water metabolism and intestinal mucosal permeability in SDDR-UC rats by inhibiting the phosphorylation of p38MAPK,ERK MAPK and up-regulating the expression of AQP4,ZO-1 and Occludin.