葛根芩连汤三氯甲烷提取物缓解伊立替康所致肠毒性的作用机制分析  

Analysis of Mechanism of Chloroform Extract of Gegen Qinliantang on Alleviating Enterotoxicity Induced by Irinotecan

作　　者：吴亿晗 成颜芬 王迪 杨晓琴 钟雪梅 林洁 傅超美[1] 章津铭[1] 胡一晨[2] WU Yi-han;CHENG Yan-fen;WANG Di;YANG Xiao-qin;ZHONG Xue-mei;LIN Jie;FU Chao-mei;ZHANG Jin-ming;HU Yi-chen(College of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;College of Food and Biological Engineering,Chengdu University,Chengdu 610106,China)

机构地区：[1]成都中医药大学药学院,成都611137 [2]成都大学食品与生物工程学院,成都610106

出　　处：《中国实验方剂学杂志》2021年第16期16-23,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：四川省科技支撑计划项目(2020YJ0195);成都中医药大学“杏林学者”学科人才科研提升计划项目;四川省科技创新苗子工程项目(2019051)。

摘　　要：目的:基于经典名方葛根芩连汤"解表清里、主协热下利"的功效,制备葛根芩连汤三氯甲烷提取物用于缓解伊立替康所致的肠毒性,为经典名方葛根芩连汤的开发与应用提供实验依据。方法:60只小鼠随机分为正常组、模型组、洛哌丁胺组(阳性药盐酸洛哌丁胺胶囊,0.4 mg·kg^(-1))和葛根芩连汤三氯甲烷提取物高、低剂量组(2.3,1.16 g·kg^(-1))。采用盐酸伊立替康(CPT-11,剂量55 mg·kg^(-1))连续4 d腹腔注射制备小鼠迟发性腹泻模型,正常组注射等体积生理盐水;分别在第5天灌胃给予相应药物,正常组和模型组给予相应体积蒸馏水,连续给药5 d。观察各组小鼠的一般情况,计算腹泻指数。运用苏木素-伊红(HE)染色观察小鼠结肠组织病理变化,按试剂盒说明书检测结肠组织中肿瘤坏死因子-α(TNF-α),白细胞介素-1β(IL-1β),环氧合酶-2(COX-2),细胞间黏附分子-1(ICAM-1),谷胱甘肽过氧化物酶(GSH-Px),超氧化物歧化酶(SOD),丙二醛(MDA),一氧化氮(NO)的含量。同时,采用蛋白免疫印迹法(Western blot)检测各组小鼠结肠组织的Kelch样环氧氯丙烷相关蛋白1(Keap1),核转录因子E2相关因子2(Nrf2),紧密连接蛋白-1(ZO-1),血红素氧合酶-1(HO-1),紧密连接蛋白(Occludin)表达情况。结果:与正常组比较,模型组经腹腔注射CPT-11造模后小鼠体质量显著降低(P<0.01),腹泻指数显著升高(P<0.01),结肠组织中TNF-α,IL-1β,COX-2,ICAM-1,MDA,NO含量显著升高(P<0.01),GSH-Px,SOD显著降低(P<0.01);HE染色显示,模型组小鼠结肠可见大量炎性细胞,结肠黏膜可见缺损、肿胀、出血及炎性渗出;结肠组织Keap1,Nrf2,ZO-1,HO-1与Occludin蛋白表达水平均显著降低(P<0.01)。与模型组比较,洛哌丁胺组和葛根芩连汤三氯甲烷提取物高、低剂量组可缓解小鼠体质量降低,且腹泻指数明显下降;TNF-α,IL-1β,COX-2,ICAM-1,MDA,NO水平降低,GSH-Px和SOD含量显著上升;HE染色显示,葛根芩连汤三氯甲烷Objective: Considering the efficacy of Gegen Qinliantang(GQT)in releasing exterior and clearing interior to alleviate dampness-heat dysentery,we analyzed the mechanism of the chloroform extract of GQT in alleviating enterotoxicity caused by irinotecan to provide an experimental basis for the development of GQT. Method:Kunming mice(n=60)were randomly divided into a blank group,a model group,a loperamide group(positive drug of loperamide hydrochloride capsule,0.4 mg·kg^(-1)),and high-(2.3 g·kg^(-1))and low-dose(1.16 g·kg^(-1))GQT chloroform extract groups. The mouse model of delayed diarrhea was established by intraperitoneal injection of irinotecan hydrochloride(CPT-11, 55 mg·kg^(-1)) for four consecutive days,meanwhile,the mice in the blank group only received the same volume of normal saline. Corresponding drugs were administered by gavage on the fifth day,respectively,while the ones in the blank group and model group were given distilled water for five consecutive days. The general condition of mice in each group was observed,and diarrhea indexes of mice were recorded. Pathological changes in colon tissues of mice were observed by hematoxylin-eosin(HE)staining. The tumor necrosis factor(TNF)-α,interleukin(IL)-1β,cyclooxygenase(COX)-2, intercellular adhesion molecule(ICAM)-1, glutathione peroxidase(GSH-Px), superoxide dismutase(SOD),malondialdehyde(MDA)and nitric oxide(NO)levels in colon tissues were detected with the assay kits. Furthermore,the expression levels of Kelch sample epoxy chloropropane associated protein 1(Keap1),nuclear factor E2 related factor 2(Nrf2),tight junction protein-1(ZO-1),heme oxygenase-1(HO-1)and tight junction protein(Occludin)were detected by Western blot. Result: Compared with the blank group,the model group showed declined body weight and reduced contents of GSH-Px and SOD(P<0.01),whereas increased diarrhea indexes and TNF-α,IL-1β,COX-2,ICAM-1,MDA and NO levels(P<0.01). Abundant inflammatory cells and colonic mucosa with defects,swelling,bleeding,and inflammatory exudation w

关 键 词：葛根芩连汤 盐酸伊立替康(CPT-11) 肠毒性 氧化应激 炎症 迟发性腹泻 洛哌丁胺 肿瘤 

分 类 号：R22[医药卫生—中医基础理论] R28[医药卫生—中医学]