盐酸小檗碱对脓毒症大鼠肠黏膜屏障损伤的保护作用及机制  

Protective effects and mechanisms of berberine hydrochloride on intestinal mucosal barrier injury in rats with sepsis

作　　者：凌颖 沈琪 王剑[2] 袁玉凡 王鹏 李泽林 晋金兰 Ling Ying;Shen Qi;Wang Jian;Yuan Yufan;Wang Peng;Li Zelin;Jin Jinlan(Department of Critical Care Medicine,Shenzhen Hospital(Futian)of Guangzhou University of Chinese Medicine,Shenzhen 518034,Guangdong,China;Department of Laboratory of Integrated Traditional Chinese and Western Medicine of Guangzhou University of Chinese Medicine,Guangzhou 510006,Guangdong,China)

机构地区：[1]广州中医药大学深圳医院(福田)重症医学科,广东深圳518034 [2]广州中医药大学中西医结合基础研究室,广东广州510006

出　　处：《中华危重病急救医学》2024年第6期597-603,共7页Chinese Critical Care Medicine

基　　金：国家自然科学基金(82274288);广东省自然科学基金(2022A1515011641);广东省深圳市科技计划项目(JCYJ20210324121808024);深圳市福田区临床重点专科项目(2024-6)。

摘　　要：目的探讨盐酸小檗碱对脓毒症大鼠肠黏膜屏障损伤的保护作用及其机制。方法将48只雄性SD大鼠按随机数字表法分为对照组(Sham组,6只)、脓毒症模型组(LPS组,14只)、盐酸小檗碱干预组(Ber组,14只)和Notch信号通路抑制组(DAPT组,14只)。DAPT组于制模前2 h腹腔注射5 mg/kg Notch信号通路抑制剂DAPT。采用向大鼠腹腔注射10 mg/kg脂多糖(LPS)的方式复制脓毒症模型;Sham组注射等量生理盐水2 mL。Ber组和DAPT组于制模2 h后灌胃50 mg/kg盐酸小檗碱;Sham组和LPS组灌胃等量生理盐水2 mL。分别于制模0、6、12、24 h观察大鼠体温、体质量、行为学和存活情况。制模24 h后麻醉取腹主动脉血,处死大鼠后取回肠组织。光镜下观察回肠组织病理学改变;透射电镜下观察回肠组织超微结构;酶联免疫吸附试验(ELISA)检测血清二胺氧化酶(DAO)、肠脂肪酸结合蛋白(iFABP)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平;实时聚合酶链反应(RT-PCR)和蛋白质免疫印迹试验(Western blotting)分别检测回肠组织紧密连接蛋白〔闭合蛋白(Occludin)、封闭蛋白1(Claudin1)〕、Notch1及其下游靶信号的mRNA和蛋白表达。结果制模后24 h,与Sham组比较,LPS组、Ber组和DAPT组大鼠体质量均下降,体温均有所升高,其中LPS组变化最明显,DAPT组次之,Ber组改变最轻;LPS组、Ber组和DAPT组存活率均较Sham组下降〔42.9%(6/14)、57.1%(8/14)、57.1%(8/14)比100%(6/6)〕,故每组取6只用于后续检测。肠道大体观察显示,LPS组大鼠回肠组织水肿及腹腔渗血情况最为严重,Ber组情况明显改善,DAPT组次之。光镜下显示,LPS组大鼠回肠黏膜腺体排列紊乱,杯状细胞明显减少,伴大量炎症细胞浸润;Ber组明显改善;DAPT组差于Ber组。电镜下显示,LPS组大鼠回肠微绒毛大量脱落,肠上皮细胞间连接复合体结构严重损伤;Ber组情况明显改善,DAPT组差于Ber组。LPS组大鼠血清DAO、iFABP、TNF-α、IL-6水�Objective To investigate the protective effect of berberine hydrochloride on intestinal mucosal barrier damage in sepsis rats and its mechanism.Methods Forty-eight male SD rats were divided into a control group(Sham group,6 cases),a sepsis model group(LPS group,14 cases),a berberine hydrochloride intervention group(Ber group,14 cases),and a Notch signaling pathway inhibition group(DAPT group,14 cases)according to random number table method.The DAPT group was intraperitoneally injected with 5 mg/kg Notch signaling pathway inhibition DAPT 2 hours before modeling.The sepsis model was established by intraperitoneal injection of 10 mg/kg lipopolysaccharide(LPS);Sham group was injected with an equal amount of saline(2 mL).The Ber group and DAPT group were treated with gavage of 50 mg/kg berberine hydrochloride 2 hours after modeling;Sham group and LPS group were treated with gavage of an equal amount of saline(2 mL).The temperature,weight,behavior and survival rate of rats were observed at 0,6,12 and 24 hours of modeling.After 24 hours of modeling,abdominal aortic blood was collected under anesthesia,and intestinal tissues were obtained after euthanasia.The pathological changes of ileum were observed under light microscope.The ultrastructure of ileum was observed under transmission electron microscope.Enzyme linked immunosorbent assay(ELISA)was used to detect the levels of serum diamine oxidase(DAO),intestinal fatty acid binding protein(iFABP),tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6).Real time-polymerase chain reaction(RT-PCR)and Western blotting were used to detect the mRNA and protein expressions of tight junction proteins(Occludin and Claudin1),Notch1 and their downstream target signals in the ileum tissue.Results After 24 hours of modeling,compared with the Sham group,the LPS group,Ber group,and DAPT group showed a decrease in weight and an increase in temperature.Among them,the LPS group showed the most significant changes,followed by the DAPT group,and the Ber group showed the least significant ch

关 键 词：盐酸小檗碱 脓毒症 肠黏膜屏障 紧密连接 Notch1信号通路 

分 类 号：R459.7[医药卫生—急诊医学]