大建中汤通过调节P2X7R介导胶质细胞活化减轻大鼠肠易激综合征内脏痛  

Dajianzhongtang attenuates visceral pain in rats with irritable bowel syn⁃drome by regulating P2X7R-mediated glial cell activation

作　　者：武静 田维毅 蔡琨 李尧锋 杨莎莎 WU jing;TIAN Weiyi;CAI Kun;LI Yaofeng;YANG Shasha(School of Basic Medicine,Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China;The First Affi-liated Hospital,Guizhou University of Traditional Chinese Medicine,Guiyang 550001,China)

机构地区：[1]贵州中医药大学基础医学院,贵州贵阳550025 [2]贵州中医药大学第一附属医院,贵州贵阳550001

出　　处：《中国病理生理杂志》2023年第6期1077-1085,共9页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金项目(No.82260855,No.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号);贵州省卫生健康委科学技术基金项目(No.gzwkj2023-503)。

摘　　要：目的:研究肠易激综合征(irritable bowel syndrome,IBS)内脏痛大鼠脊髓后角(posterior horn of spinal cord,PHSC)和前扣带回皮质(anterior cingulate cortex,ACC)星形胶质细胞中胶质细胞原纤维酸性蛋白(glial fibrillary acidic protein,GFAP)及P2X7受体(P2X7 receptor,P2X7R)表达的变化及大建中汤(Dajianzhongtang,DJZT)的干预作用。方法:通过腹腔注射卵清白蛋白、母婴分离、乙酸灌肠等方式制备IBS内脏痛大鼠模型,将模型大鼠随机分为3组:模型组(灌服等体积生理盐水,每天1次,持续2周)、DJZT组(10.8 g/kg DJZT灌胃,每天1次,持续2周)和P2X7R拮抗剂亮蓝G(brilliant blue G,BBG)组(腹腔注射50 mg/kg,每天1次,持续2周),另设正常组(灌服等体积生理盐水,每天1次,持续2周),共计4组,每组10只。采用腹部撤离反射(abdominal withdrawal reflex,AWR)评估各组大鼠内脏疼痛敏感度;苏木精-伊红(hematoxylin-eosin,HE)染色观察大鼠结肠黏膜的病理变化;RT-qPCR和Western blot检测大鼠PHSC和ACC组织中脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)、白细胞介素1β(interleukin-1β,IL-1β)、GFAP和P2X7R的mRNA及蛋白表达水平;免疫荧光双染法观察PHSC和ACC组织中GFAP和P2X7R的定位情况。结果:与正常组大鼠比较,模型组大鼠AWR评分于60、40和20 mmHg压力下增加(P<0.01),结肠组织HE染色病变均未见病理改变,PHSC和ACC组织中BDNF、IL-1β、GFAP和P2X7R的mRNA及蛋白表达水平显著升高(P<0.01);PHSC和ACC组织中GFAP与P2X7R共表达,且模型组PHSC和ACC组织中GFAP和P2X7R共表达阳性细胞数量显著增多(P<0.01)。与模型组相比,BBG组和DJZT组AWR评分于60、40和20 mmHg压力下下降(P<0.01),ACC和PHSC组织中IL-1β、BDNF、GFAP和P2X7R的mRNA及蛋白表达水平显著降低(P<0.01);BBG组大鼠PHSC和ACC组织中GFAP和P2X7R共表达阳性细胞数量显著减少(P<0.01)。DJZT组和BBG组上述各指标的差异均无统计学意义。结论:DJZT减轻大鼠IBS内脏痛的机制可�AIM:To explore the mechanism of Dajianzhongtang(DJZT)in treating visceral pain in rats with irritable bowel syndrome(IBS).METHODS:The IBS visceral pain rat model was prepared by intraperitoneal injection of oval albumin,mother-infant separation,acetic acid gavage,etc.The model rats were randomly divided into 3 groups:model group(instillation of equal volume of normal saline,once a day for 2 weeks),DJZT group(gavage of 10.8 g/kg DJZT,once a day for 2 weeks)and P2X7 receptor(P2X7R)antagonist brilliant blue G(BBG)group(intraperitoneal injection of 50 mg/kg BBG,once a day for 2 weeks).Another control group(gavage of equal volume of normal saline,once a day for 2 weeks)was set up.There were a total of 4 groups,with 10 rats in each group.The abdominal withdrawal reflex(AWR)was used to assess the visceral pain sensitivity of each group of rats.Hematoxylin-eosin(HE)staining was used to observe the pathological changes in the colonic mucosa of rats.The mRNA and protein expression levels of brain-derived neurotrophic factor(BDNF),interleukin-1β(IL-1β),glial fibrillary acidic protein(GFAP)and P2X7 receptor(P2X7R)in posterior horn of spinal cord(PHSC)and anterior cingulate cortex(ACC)tissues were measured by RT-qPCR and Western blot.The localization of GFAP and P2X7R in PHSC and ACC tissues was observed by immunofluorescence dou‐ble-staining.RESULTS:Compared with control group,AWR scores of the rats in model group were significantly in‐creased at 60,40 and 20 mmHg pressure(P<0.01).No significant pathological changes were seen in HE-stained lesions of colonic tissues.The mRNA and protein expression levels of BDNF,IL-1β,GFAP and P2X7R were significantly in‐creased in PHSC and ACC tissues(P<0.01).The GFAP and P2X7R were co-expressed in PHSC and ACC tissues,and the number of GFAP and P2X7R co-expressing positive cells increased in PHSC and ACC tissaes(P<0.01).Compared with model group,AWR scores of the rats in BBG and DJZT groups significantly decreased at 60,40 and 20 mmHg pres‐sure(P<0.01),and the IL-1β,BDNF,GFAP a

关 键 词：大建中汤 肠易激综合征 内脏痛 胶质细胞原纤维酸性蛋白 P2X7受体 星形胶质细胞 

分 类 号：R285[医药卫生—中药学] R363[医药卫生—中医学]