醒脾胶囊调节5-HT-RhoA/ROCK2通路干预脾虚腹泻型肠易激综合征大鼠的机制  

Mechanism of Intervening with Diarrhea-predominant Irritable Bowel Syndrome in Rats with Spleen Deficiency by Xingpi Capsules Through Regulating 5-HT-RhoA/ROCK2 Pathway

作　　者：王港 崔伶雯 刘相凝 朱镕鑫 黄明月 孙颖 焦伯阳 王然 李春 WANG Gang;CUI Lingwen;LIU Xiangning;ZHU Rongxin;HUANG Mingyue;SUN Ying;JIAO Boyang;WANG Ran;LI Chun(School of Traditional Chinese Medicine,School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学中医学院,中药学院,北京100029

出　　处：《中国实验方剂学杂志》2025年第6期60-69,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(8222200821);北京中医药大学校企合作项目(JXHT-202012-011)。

摘　　要：目的:探讨醒脾胶囊(XPC)在治疗脾虚腹泻型肠易激综合征(IBS-D)中的功效,并阐明其潜在的分子机制。方法:采用番泻叶给药联合约束应激和游泳疲劳连续14 d制备大鼠脾虚IBS-D模型。取10只SPF级健康大鼠作为正常组;将造模成功后的SPF级大鼠随机分为模型组、匹维溴铵组(1.5 mg·kg^(-1))和XPC低、高剂量组(0.135、0.54 g·kg^(-1)),每组10只。正常组与模型组大鼠灌胃蒸馏水,其余各组灌胃相应药液,每天1次,连续14 d。每日观测大鼠体质量和粪便情况,记录粪便Bristol评分;采用酶联免疫吸附测定法(ELISA)检测血清和结肠组织中5-羟色胺(5-HT)水平;利用透射电子显微镜观察结肠微绒毛和紧密连接等细微结构;通过苏木素-伊红(HE)染色法、免疫组织化学法和蛋白免疫印迹法(Western blot)评价结肠屏障完整性、检测肠道运动及相关通路蛋白的表达情况。结果:与正常组比较,模型组大鼠体质量下降和稀便率、稀便级及Bristol评分均显著升高(P<0.01),HE染色结果模型组见结肠黏膜层明显充血、水肿,提示其不完整,电镜结果提示结肠屏障结构致密度和完整性下降,微绒毛脱落与消失,紧密连接处发生明显增宽,结肠组织膜整合蛋白(Occludin)、紧密连接蛋白-5(Claudin-5)表达显著下调(P<0.01),血清和结肠组织中5-HT含量显著升高(P<0.01),小肠推进率显著上升(P<0.01),结肠组织Ras同源物基因家族A(RhoA)、Rho相关卷曲螺旋激酶2(ROCK2)表达及肌球蛋白轻链(MLC_(20))的磷酸化程度均显著上调(P<0.01)。与模型组比较,各治疗组腹泻及其伴随症状和结肠组织的病理表现有不同程度缓解;其中,高剂量的XPC可有效缓解腹泻并促进结肠黏膜结构恢复整齐致密同时有效减轻充血水肿,显著上调Occludin、Claudin-5表达(P<0.01),明显降低血清和结肠组织中5-HT含量(P<0.05,P<0.01),显著减缓小肠推进率(P<0.01),明显下调结肠组织收缩蛋白RhoA、ROCK2表Objective:To investigate the efficacy of Xingpi capsules(XPC)in treating diarrhea-predominant irritable bowel syndrome(IBS-D)with spleen deficiency and elucidate its potential molecular mechanisms.Methods:A rat model of IBSD with spleen deficiency was established by administering senna leaf in combination with restrained stress and swimming fatigue for 14 d.Ten specific pathogen free(SPF)-grade healthy rats were used as the normal control group.After successful modeling,SPFgrade rats were randomly divided into a model group,a pinaverium bromide group(1.5 mg·kg^(-1)),and low-and high-dose XPC groups(0.135 and 0.54 g·kg^(-1)),with 10 rats in each group.Rats in the normal control group and the model group were given distilled water by gavage,while the remaining groups were administered corresponding drug solutions by gavage once a day for 14 consecutive days.The rat body weights and fecal condition were observed every day,and the Bristol score was recorded.Enzymelinked immunosorbent assay(ELISA)was used to determine the levels of 5-hydroxytryptamine(5-HT)in serum and colon tissue.Transmission electron microscopy was used to observe the microvilli and tight junctions in the colon.The integrity of the colonic barrier,intestinal motility,and expression of related pathway proteins were evaluated by hematoxylin-eosin(HE)staining,immunohistochemistry,and Western blot.Results:Compared with those in the normal control group,rats in the model group showed a significantly decreased body weight and increased diarrhea rate,diarrhea grade,and Bristol score(P<0.01).HE staining revealed incomplete colonic mucosa in the model group,with evident congestion and edema observed.Electron microscopy results indicated decreased density and integrity of the colonic barrier,shedding and disappearance of microvilli,and significant widening of tight junctions.The expression levels of colonic tight junction proteins Occludin and Claudin-5 were downregulated(P<0.01),and the levels of 5-HT in serum and colon tissue were elevated(P<0.01).The sm

关 键 词：腹泻型肠易激综合征 醒脾胶囊 5-羟色胺(5-HT)-Ras同源物基因家族A(RhoA)/Rho相关卷曲螺旋激酶2(ROCK2)2途径 

分 类 号：R242[医药卫生—中医临床基础] R289[医药卫生—中医学] R318.14