基于“肠脑互动”探讨大建中汤治疗肠易激综合征大鼠内脏痛作用机制  

Mechanism of Dajianzhong Decoction(大建中汤)Treating Visceral Pain in Rats with IBS Based on“Intestine-Brain Interaction”

作　　者：武静 王慧 李尧锋 王俊霞 杨毅 陈刚[1] WU Jing;WANG Hui;LI Yaofeng;WANG Junxia;YANG Yi;CHEN Gang(Hubei University of Chinese Medicine,Wuhan 430065,Hubei,China;Guizhou University of Traditional Chinese Medicine,Guiyang 550025,Guizhou,China)

机构地区：[1]湖北中医药大学,湖北武汉430065 [2]贵州中医药大学,贵州贵阳550025

出　　处：《中华中医药学刊》2024年第8期102-106,I0016,I0017,I0018,共8页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(82260855,81960821);贵州省基础研究计划(自然科学)项目(黔科合基础-ZK[2022]一般495);贵州省卫生健康委员会科学技术基金项目(gzwkj2023-503)。

摘　　要：目的研究大建中汤对肠易激综合征(irritable bowel syndrome,IBS)大鼠内脏痛的影响及神经-免疫-内分泌网络的分子机制。方法将新生大鼠48只,随机分为正常对照组(control组)、肠易激综合征模型组(IBS组)、酮替芬组(Ketotifen组)、低剂量大建中汤治疗组(DJZT-L)、中剂量大建中汤治疗组(DJZT-M)、高剂量大建中汤治疗组(DJZT-H)。采用母婴分离法制备IBS内脏痛大鼠模型,利用腹壁撤退反应(abdominal withdrawal reflex,AWR)评估大鼠内脏敏感性;免疫组化法测定大鼠下丘脑星形胶质细胞(glial fibrillary acidic protein,GFAP)、小胶质细胞(OX-42);免疫蛋白印迹(Western blot,WB)检测结肠和下丘脑P物质(substance P,SP);甲苯胺蓝染色法检测结肠组织肥大细胞(mast cell,MC)脱颗粒率;酶联免疫吸附法(enzyme-linked immunosorbentassay,ELISA)测定分析结肠组织SP、特异性IgE(sIgE)和组胺。结果与正常大鼠相比,模型大鼠中60、40和20 mm Hg压力下AWR评分明显升高(P<0.05),下丘脑GFAP、OX-42、SP表达显著上升(P<0.05),结肠组织中肥大细胞脱颗粒率、sIgE、组胺和SP表达显著上升(P<0.05);与模型大鼠相比,大建中汤高剂量组(40和20 mm Hg压力),大建中汤中剂量组(60、40和20 mm Hg压力)的AWR评分,下丘脑GFAP、OX-42、SP,结肠肥大细胞脱颗粒率、sIgE、组胺、SP表达明显下降(P<0.05)。结论大建中汤能够降低IBS模型大鼠内脏痛,可能与其抑制SD大鼠结肠组织中肥大细胞活性,进而影响免疫神经网络有关。Objective To study the effects of Dajianzhong Decoction(大建中汤,DJZD)on visceral pain in rats with irritable bowel syndrome(IBS)and the molecular mechanism of neuroimmune-endocrine network.Methods Forty-eight newborn rats are randomly divided into normal control group,IBS model group,Ketotifen group and DJZD low-dose,medium-dose and high-dose groups(DJZD-L,DJZD-M,DJZD-H).The rat model of IBS visceral pain was prepared by mother-infant separation method,and abdominal withdrawal reflex(AWR)was used to evaluate the visceral sensitivity of rats.Astrocytes(GFAP)and microglia cells(OX-42)were determined by immunohistochemistry in the hypothalamus.Substance P(SP)of colon and hypothalamus was detected by Western blot.The degranulation rate of colon mast cells was determined by toluidine blue staining.Colon tissue-specific IgE(sIgE),Histamine(Histamine)and Substance P(SP)were analyzed by enzymedy-linked immunosorbent assay(ELISA).Results Compared with those of the normal rats,AWR score of model rats under pressure of 60,40 and 20 mm Hg significantly increased(P<0.05),the expressions of GFAP,OX-42 and SP in hypothalamus significantly increased(P<0.05),mast cell degranulation rate,sIgE,histamine and SP in colon tissue significantly increased(P<0.05).Compared with those of the model rats,AWR scores,GFAP,OX-42 and SP in hypothalamus,mast cell degranulation rate,sIgE,histamine and SP expression in colon significantly decreased in DJZD-H group(40 and 20 mm Hg pressures)and DJZD-M group(60,40 and 20 mm Hg pressures)(P<0.05).Conclusion DJZD can reduce visceral pain in IBS model rats,which may be related to inhibiting the activity of mast cells in colon tissue of SD rats to affect the immune neural network.

关 键 词：肠易激综合征 内脏痛 肥大细胞 大建中汤 胶质细胞 

分 类 号：R289.5[医药卫生—方剂学]