小陷胸汤对5-氟尿嘧啶介导的小鼠肠黏膜损伤的保护作用研究  

Study on the protective effect of Xiaoxianxiong decoction on 5⁃fluorouracil⁃induced intestinal damage in mice

作　　者：秦美洁 叶倩倩 丁芮 黄金玲[2,3] QIN Meijie;YE Qianqian;DING Rui;HUANG Jinling(School of Traditional Chinese Medicine,Anhui University of Traditional Chinese Medicine,Hefei 230012,China;School of Integrated Traditional Chinese and Western Medicine,Anhui University of Traditional Chinese Medicine,Hefei 230012,China;Anhui Province Key Laboratory of Compound Traditional Chinese Medicine,Hefei 230012,China)

机构地区：[1]安徽中医药大学中医学院,安徽合肥230012 [2]安徽中医药大学中西医结合学院,安徽合肥230012 [3]中药复方安徽省重点实验室,安徽合肥230012

出　　处：《海南医学院学报》2024年第19期1441-1450,共10页Journal of Hainan Medical University

基　　金：国家自然科学基金资助项目(81973844、81573864)。

摘　　要：目的:探究小陷胸汤对5-氟尿嘧啶(5-fluorouracil,5-FU)介导的小鼠肠黏膜损伤的保护作用。方法:将100只SPF级雄性ICR小鼠随机分为正常对照组、模型组、小陷胸汤低、中、高剂量(8.3、16.6、33.2g/kg)组。除正常对照组外,其余各组采用5-FU(30 mg/kg,ip,7 d)诱导肠黏膜损伤小鼠模型。小陷胸汤给药组造模同时按照设定剂量以10 mL·kg^(-1)·d^(-1)灌胃,正常对照组和模型组给予等量蒸馏水,连续处理7 d。采用苏木素-伊红染色法(HE)观察小肠的病理形态学变化并测定肠绒毛高度/隐窝深度(VH:CD)比值;采用阿利新蓝染色法(AB)测定小肠黏膜杯状细胞数量;采用分光光度法检测血清内毒素(ET)、D-乳酸(D-LA)及小肠组织二胺氧化酶(DAO)水平;采用酶联免疫吸附法(ELISA)检测血清肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)水平;采用实时荧光定量聚合酶链式反应法(RT-qPCR)检测小肠组织IL-6、信号转导和转录激活因子3(STAT3)和表皮生长因子受体(EGFR)mRNA表达;采用Western blot法检测小肠组织IL-6、STAT3及EGFR蛋白表达。结果:与正常对照组相比,模型组小鼠体质量显著降低,腹泻评分、粪便质量、粪便含水量及小肠推进率明显升高(P<0.01),小肠黏膜VH:CD比值和杯状细胞数量降低(P<0.01),血清ET和D-LA水平升高,小肠组织DAO水平降低(P<0.05),脾指数和胸腺指数降低(P<0.01),血清IL-6、TNF-α水平升高(P<0.05),小肠组织IL-6、STAT3 mRNA表达升高,EGFR mRNA表达降低(P<0.01);小肠组织IL-6、STAT3蛋白表达升高,EGFR蛋白表达降低(P<0.01)。与模型组相比,小陷胸汤能够明显改善上述指标(P<0.05)。结论:小陷胸汤对5-FU介导的肠黏膜损伤具有保护作用,其作用机制可能与小陷胸汤激活EGFR信号转导,下调IL-6/STAT3信号通路,减轻5-FU诱导的肠黏膜炎症反应,改善免疫功能有关。Objective:To investigate how Xiaoxianxiong decoction can protect against intestinal damage caused by 5⁃fluoroura⁃cil(5⁃FU)in mice.Methods:One hundred male ICR breed mice with SPF⁃grade were randomised into the normal control group,the model group and the low,medium and high dose(8.3,16.6 and 33.2 g/kg)groups of Xiaoxianxiong decoction.Intestinal mu⁃cosal injury was induced in all groups,except the control group,by administering 5⁃FU(30 mg/kg,ip,7 d).While modeling was ongoing,the mice in the Xiaoxianxiong decoction group were administered 10 mL·kg^(⁃1)·d^(⁃1)via gavage,following the prescribed dos⁃age.The control and model groups were administered equal volume of distilled water and treated continuously for 7 d.Histopatho⁃logical alterations in the small intestine were assessed utilizing hematoxylin⁃eosin staining(HE),with the villus height to crypt depth ratio(VH:CD)subsequently determined.The number of goblet cells in the small intestinal mucosa was determined by Al⁃cian blue staining(AB).The concentrations of endotoxin(ET)and D⁃lactic acid(D⁃LA)in serum were measured using spectro⁃photometry,along with diamine oxidase(DAO)levels in the small intestine.The levels of interleukin⁃6(IL⁃6)and tumor necro⁃sis factor⁃α(TNF⁃α)in serum were detected using enzyme⁃linked immunosorbent assay(ELISA).The mRNA expression of IL⁃6,signal transducer and activator of transcription 3(STAT3),and epidermal growth factor receptor(EGFR)was detected us⁃ing reverse transcription quantitative polymerase chain reaction(RT⁃qPCR).The IL⁃6,STAT3 and EGFR protein were expres⁃sion in small intestinal tissues were detected by the Western blot method.Results:Compared to the control group,the model group of mice presented a significant reduction in body mass and an increase in scores for diarrhoea,faecal mass,faecal water con⁃tent,and small intestinal propulsion rates.(P<0.01).The VH:CD ratio,as well as the number of goblet cells in the small intesti⁃nal mucosa,exhibited a noteworthy de

关 键 词：小陷胸汤 5-氟尿嘧啶(5-FU) 肠黏膜损伤 表皮生长因子受体(EGFR) 白细胞介素6(IL-6)/信号转导和转录激活因子3(STAT3)信号通路 

分 类 号：R285.5[医药卫生—中药学]