醒脾胶囊调控血管活性肠肽干预功能性腹泻脾虚证大鼠机制研究  

Mechanism of Xingpi Capsule(醒脾胶囊)to Modulate Vasoactive Intestinal Peptide Intervening in Model Rats with Spleen-Deficiency Syndrome Functional Diarrhea

作　　者：王港 崔伶雯 刘相凝 刘一舟 千维娜 胡玥瑶 邓琼[1] 董健藤 孙小仟 李俊俊 李春[1] 王勇[1] WANG Gang;CUI Lingwen;LIU Xiangning;LIU Yizhou;QIAN Weina;HU Yueyao;DENG Qiong;DONG Jianteng;SUN Xiaoqian;LI Junjun;LI Chun;WANG Yong(Beijing University of Chinese Medicine,Beijing 102488,China)

机构地区：[1]北京中医药大学,北京102488

出　　处：《辽宁中医药大学学报》2023年第3期19-26,共8页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(82174215)。

摘　　要：目的探讨醒脾胶囊通过血管活性肠肽(vasoactive intestinal peptide,VIP)修复肠道屏障且调节RhoA/ROCK2信号通路抑制肠道运动亢进,缓解功能性腹泻(functional diarrhea,FDr)脾虚证的作用机制。方法将50只SPF级大鼠按照随机数字表均分成正常组,模型组,洛哌丁胺组和醒脾胶囊高、低剂量组。采用高浓度番泻叶水煎剂连续灌胃14 d制备脾虚型FDr膜型。依据临床脾虚泄泻诊断标准,动态采集并量化排便情况、体质量、饮食量、疲劳程度、皮毛情况等大鼠宏观体征;28 d测定小肠推进率并取结肠组织和腹主动脉血;采用酶联免疫吸附试验测定血清和结肠组织中VIP含量,通过检测苏木精-伊红(HE)染色法和蛋白质印迹法评价结肠屏障完整性、肠道运动及相关通路蛋白的表达情况。结果与正常组相比,14 d后,模型组大鼠便多稀溏,体质量增长减缓,喜眯眼蜷卧,毛枯少泽,食少乏力,量化后各项指标评分及总分显著升高(P<0.01),小肠推进率明显上升(P<0.01),血清和结肠组织中VIP含量升高(P<0.01)。HE染色提示模型组结肠黏膜层不完整,可见明显充血、水肿,结肠紧密连接蛋白Occludin和Claudin-5表达下调(P<0.01),结肠收缩蛋白RhoA和ROCK2表达均上调(P<0.01,P<0.05)。与模型组相比,各治疗组大鼠腹泻及其伴随症状和结肠组织切片的病理表现有不同程度缓解;其中,高剂量的醒脾胶囊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和Claudin-5表达(P<0.01),降低血清和结肠组织中VIP含量(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和ROCK2表达(P<0.01)。结论醒脾胶囊可有效缓解大鼠脾虚症状并改善腹泻,调控VIP从上调肠屏障紧密连接与抑制肠道过亢运动两个方面达到缓解脾虚型FDr的功效。Objective To explore the potential mechanism of Xingpi Capsule(醒脾胶囊,XPC)to restore intestinal barrier and regulate RhoA/ROCK2 signaling pathway by inhibiting intestinal hypermotility via vasoactive intestinal peptide(VIP)and alleviate functional diarrhea(FDr)with spleen-deficiency syndrome.Methods Fifty specific pathogen free rats were divided into normal,model,loperamide,and XPC high and low dose groups by a random number table.The model of spleendeficiency syndrome FDr was prepared by gavaging with a high concentration of senna leaf decoction for 14 d.According to the diagnostic criteria of the spleen-deficiency syndrome diarrhea in clinic,signs such as defecation,body mass,food intake,fatigue,and skin condition were collected and quantified.On day 28,small intestine propulsion was measured.VIP levels in serum and colonic tissues were measured by enzyme-linked immunosorbent assay.The colonic barrier integrity and intestinal motility,and related protein expression were assessed by hematoxylin-eosin staining(HE)and Western blotting.Results Compared with the normal group,the rats in the model group had more loose stools,slower body mass growth,squinting and curling up,less lustrous hair,less food intake and weakness.The score of each index and total score were significantly increased(P<0.01).The small intestine propulsion in the model group was increased(P<0.01).The VIP levels in the serum and colon tissues were increased(P<0.01).HE staining showed incomplete mucosal layer of the colon,swelling,and edema in the model group.The expression of the tight junction proteins Occludin and Claudin-5 was downregulated(P<0.01)and the expression of contractile proteins RhoA and ROCK2 in the colon tissues was upregulated(P<0.01 and P<0.05).Compared with the model group,the treatment groups showed different degrees of alleviation of diarrhea,its accompanying symptoms,and pathological manifestations of sliced colon tissues.High dose of XPC effectively alleviated diarrhea,rearranged the structure of intestinal mucosa tigh

关 键 词：功能性腹泻 脾虚证 醒脾胶囊 血管活性肠肽 Occludin CLAUDIN-5 RhoA/ROCK2通路 

分 类 号：R574.62[医药卫生—消化系统]