健脾解毒方调控TLRs/NF-κB信号通路对5-FU致大鼠肠黏膜炎的保护作用  

Jianpi Jiedu Formula alleviated 5-FU-induced intestinal mucositis in rats through inhibiting TLRs/NF-κB signaling pathway

作　　者：刘慧[1] 冯丹青[1] 邵诗芸 贾茹 张影茹 程悦蕾[1] 张彦博[1] 赖优莹[1] 占向阳 张必萌[4] 李琦[1,2,3] 王炎[1,2,3] LIU Hui;FENG Danqing;SHAO Shiyun;JIA Ru;ZHANG Yingru;CHENG Yuelei;ZHANG Yanbo;LAI Youying;ZHAN Xiangyang;ZHANG Bimeng;LI Qi;WANG Yan(Department of Oncology,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Cancer Institute,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Academy of Integrative Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Department of Acupuncture and Moxibusdon,Shanghai General Hospital,Shanghai Jiao Tong University,Shanghai 201620,China)

机构地区：[1]上海中医药大学附属曙光医院肿瘤科,上海201203 [2]上海中医药大学附属曙光医院肿瘤研究所,上海201203 [3]上海中医药大学中西医结合研究院,上海201203 [4]上海交通大学附属第一人民医院针灸科,上海201620

出　　处：《上海中医药杂志》2020年第12期84-90,共7页Shanghai Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81830120,81703887,82074232,81774095);上海市科委科研计划项目(19401972200);上海市中医药事业发展三年行动计划资助项目[ZY(2018-2020)-CCCX-2003-03];上海中医药大学预算内项目(2015YSN41)。

摘　　要：目的观察健脾解毒方通过TLRs/NF-κB信号通路对5-FU致大鼠肠黏膜炎的保护作用。方法将60只SD大鼠随机分为空白组、模型组、健脾解毒方低、中、高剂量组(0.31 g/mL,0.62 g/mL,1.24 g/mL)、培菲康组6组,每组10只。除空白组外,每组大鼠予5-FU(50 mg/kg)腹腔注射,每日1次,连续5 d;中药治疗组在化疗的同时分别灌胃给予低、中、高剂量健脾解毒方(3.1 g/kg,6.2 g/kg,12.4 g/kg),每天1次,连续7 d;培菲康组在化疗的同时灌胃给予培菲康(151 mg/kg),每天1次,连续7 d。每日观察大鼠腹泻情况、测量大鼠体质量,7天后HE染色观察大鼠小肠组织结构,免疫组化检测小肠组织中紧密连接蛋白Occludin蛋白的表达,ELISA法检测各组大鼠血清TNF-α、IL-10表达,Western blot法检测肠组织中MyD88、p65与TLRs含量。结果 (1)实验过程中空白组大鼠体质量进行性增加,实验第3天起模型组及各给药组大鼠体质量开始下降,实验第5天起,健脾解毒方高剂量组大鼠体质量下降小于模型组(P<0.05);第5天起,除空白组外的其他组大鼠出现腹泻,至第6天达到腹泻高峰,且第6天健脾解毒方高剂量组的腹泻发生率低于模型组(P<0.05)。(2)与空白组比较,模型组Occludin蛋白表达量减少(P<0.01);与模型组比较,健脾解毒方各剂量组Occludin蛋白表达量均增加,其中以健脾解毒方高剂量组增加最显著(P<0.01)。(3)模型组大鼠血清TNF-α表达高于空白组(P<0.05),各治疗组的TNF-α均低于模型组,且健脾解毒方中、高剂量组的TNF-α低于健脾解毒方低剂量组(P<0.05);模型组大鼠的IL-10低于空白组(P<0.05),健脾解毒方高剂量组、培菲康组高于模型组(P<0.05)。(4)模型组MyD88、p65、TLR4蛋白表达高于空白组(P<0.01),健脾解毒方中、高剂量组、培菲康组低于模型组,且健脾解毒方中、高剂量组的蛋白表达低于健脾解毒方低剂量组(P<0.01)。结论健脾解毒方能够抑制TLRs/NF-κB信号通路,调节炎Objective To observe the protective effect of Jianpi Jiedu Formula(JPJDF)on 5-FU-induced intestinal mucositis in rats through inhibiting TLRs/NF-κB signaling pathway. Methods Sixty SD rats were randomly divided into blank group, model group, JPJDF-Low(0.31 g/mL), JPJDF-Medium(0.62 g/mL) and JPJDF-High(1.24 g/mL) groups of(0.31 g/mL, 0.62 g/mL, 1.24 g/mL)and Bifico group, with 10 rats in each group. All the rats except the rats in the blank group were injected intraperitoneally with 5-FU(50 mg/kg) once a day for 5 consecutive days. Rats in Chinese medicine treatment groups were given low, middle and high dose of JPJDF(3.1 g/kg, 6.2 g/kg, 12.4 g/kg)respectively, once a day for 7 days during the chemotherapy, while those in Bifico group were given Bifico(151 mg/kg)daily for 7 days. Diarrhea and body weight of rats were recorded daily. Seven days later, HE staining was used to observe the structure of rat small intestine. Immunohistochemistry was used to detect the expression of Occludin protein in small intestine.The serum levels of TNF-α and IL-10 were measured by ELISA, and the expressions of MyD88, p65 and TLRs in the intestine were detected by Western blot. Results(1)During the experiment, the weight of the rats in the blank group increased progressively. From the 3 rd day, the weight of the rats in the model, JPJDF-Low, JPJDF-Medium, JPJDF-High and Bifico groups began to decrease. From the 5 th day of the experiment, the rats in the JPJDF-High group had milder weight loss compared with those in the model group(P<0.05). Diarrhea occurred in all groups except the blank group from the 5 th day and reached the peak on the 6 th day, and the incidence of diarrhea in the JPJDF-High group was lower than that in the model group on the 6 th day(P<0.05).(2)Compared with the blank group, the expression of Occludin protein in the model group decreased(P<0.01). Compared with the model group, the expression of Occludin protein in all dose groups of JPJDF increased, among which the JPJDF-High group increased most significant

关 键 词：肠黏膜炎 大鼠 5-氟尿嘧啶 健脾解毒方 信号通路 

分 类 号：R285.5[医药卫生—中药学]