摘要
目的:探讨四神丸改善腹泻型肠易激综合征大鼠的内脏敏感性的作用及其可能机制。方法:40只雄性SPF级大鼠随机分为空白组、模型组、四神丸低、高剂量组(3.51、7.02 g·kg^(-1))、培菲康组(0.54 g·kg^(-1))。除空白组外,其余均以母子分离刺激+番泻叶水煎剂灌胃构建脾肾阳虚型腹泻型肠易激综合征大鼠模型。给予相应药物干预后:观察大鼠一般情况,测量大鼠6 h排便颗粒及腹壁撤退反射实验(AWR)最小容量阈值,酶联免疫吸附测定法检测大鼠血清肿瘤坏死因子-α(TNF-α)、胃泌素(GAS)、皮质酮(CORT)、促肾上腺皮质激素(ACTH)含量,苏木素-伊红(HE)染色法观察大鼠结肠组织形态,甲苯胺蓝染色法观察大鼠结肠组织肥大细胞脱颗粒情况,蛋白免疫印迹法(Western blot)检测大鼠结肠组织p38丝裂原活化蛋白激酶(p38 MAPK)、c-Jun氨基末端激酶(JNK)、瞬时感受器电位香草酸受体1(TRPV1)、蛋白酶激活受体2(PAR2)蛋白表达,免疫组化法检测结肠组织TRPV1蛋白含量,免疫荧光法检测结肠组织神经肽P物质(SP)和降钙素基因相关肽(CGRP)蛋白阳性表达。结果:与空白组比较,模型组大鼠6 h排便颗粒显著增加(P<0.01),AWR最小容量阈值显著降低(P<0.01),TNF-α含量显著升高(P<0.01),GAS、CORT、ACTH含量明显降低(P<0.05,P<0.01),肥大细胞脱颗粒率升高(P<0.01),TRPV1的阳性表达明显升高(P<0.05),SP、CGRP蛋白阳性表达明显升高(P<0.05,P<0.01),p38 MAPK、JNK、TRPV1、PAR2蛋白表达量均显著升高(P<0.01);与模型组比较,四神丸高剂量组AWR最小容量阈值显著升高(P<0.01),四神丸高剂量组、培菲康组排便频率明显下降(P<0.05,P<0.01),TNF-α含量明显降低(P<0.05,P<0.01),GAS、CORT、ACTH含量明显升高(P<0.05,P<0.01),肥大细胞脱颗粒率下降(P<0.01),TRPV1阳性表达下降(P<0.05),SP、CGRP表达水平明显降低(P<0.05,P<0.01),p38 MAPK、JNK、TRPV1、PAR2蛋白表达量明显降低(P<0.05,P<0.01)。结论:四�
Objective:To investigate the effect and possible mechanism of Sishenwan in ameliorating visceral sensitivity in the rat model of diarrhea-predominant irritable bowel syndrome(IBS-D)due to spleenkidney Yang deficiency.Method:Forty male SPF-grade rats were randomly assigned into five groups:blank control,model,low-(3.51 g·kg^(-1))and high-dose(7.02 g·kg^(-1))Sishenwan,and Peifikang(0.54 g·kg^(-1))groups.Except the blank control group,the other groups underwent maternal separation stress and Sennae Folium decoction gavage for the modeling of IBS-D due to spleen-kidney Yang deficiency.After corresponding drug interventions,the general conditions of the rats were observed,and the number of defecation pellets within 6 h and the minimum threshold of abdominal withdrawal reflex(AWR)were measured.Enzyme-linked immunosorbent assay(ELISA)was used to measure the serum levels of tumor necrosis factor(TNF)-α,gastrin(GAS),corticosterone(CORT),and adrenocorticotropic hormone(ACTH).Hematoxylin-eosin(HE)staining was employed to observe pathological changes in the colon tissue.Toluidine blue staining was used to assess mast cell degranulation in the colon tissue.Western blot was performed to determine the protein levels of p38 mitogen-activated protein kinase(MAPK),c-Jun N-terminal kinase(JNK),transient receptor potential vanilloid 1(TRPV1),and protease-activated receptor 2(PAR2)in the colon tissue.Immunohistochemistry was employed to measure the protein level of TRPV1 in the colon tissue,and immunofluorescence was used to detect the positive expression of substance P(SP)and calcitonin gene-related peptide(CGRP)in the colon tissue.Result:Compared with the blank control group,the model group showed increased number of defecation pellets within 6 h(P<0.01),decreased minimum threshold of AWR(P<0.01),elevated serum TNF-αlevel(P<0.01),lowered levels of GAS,CORT,and ACTH(P<0.05,P<0.01),increased mast cell degranulation rate(P<0.01),increased positive expression of TRPV1,SP,and CGRP(P<0.05,P<0.01),and upregulated protein levels of p3
作者
黎思琪
胡运莲
苏成霞
萧闵
江晓翠
文娜
章茜
LI Siqi;HU Yunlian;SU Chengxia;XIAO Min;JIANG Xiaocui;WEN Na;ZHANG Qian(College of Traditional Chinese Medicine(TCM),Hubei University of Chinese Medicine,Wuhan 430061,China;Hubei Shizhen Laboratory,Wuhan 430061,China;First Clinical College,Hubei University of Chinese Medicine,Wuhan 430061,China;Hubei Provincial Hospital of TCM,Affiliated Hospital of Hubei University of Chinese Medicine,Wuhan 430060,China;Laboratory Animal Center,Hubei University of Chinese Medicine,Wuhan 430065,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2024年第21期10-18,共9页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金面上项目(82374205)
湖北省中医药管理局2023—2024年度中医药青年人才项目(ZY2023Q039)
湖北省中医药管理局2023—2024年度中医药指导性项目(ZY2023F004)
武汉市科技局的曙光项目(2022020801020508)。
