世界中医药
文章摘要
引用本文:冷静1,2,田华捷1,2,黄甫1,2,冯琴1,2,赵瑜1,2,胡义扬1,3,4,彭景华1,2,3.BZL方对非酒精性脂肪性肝炎模型小鼠肠黏膜损伤的保护作用[J].世界中医药,2019,(03):.  
BZL方对非酒精性脂肪性肝炎模型小鼠肠黏膜损伤的保护作用
Protective Effects of BZL Recipe on Intestinal Mucosal Injury in Non-alcoholic Steatohepatitis in Mice
投稿时间:2018-11-12  
DOI:10.3969/j.issn.1673-7202.2019.03.014
中文关键词:  非酒精性脂肪性肝炎  中药组分复方  天然产物  白术多糖  栀子苷  绿原酸  肠黏膜屏障  紧密连接
English Keywords:Non-alcoholic steatohepatitis  Chinese medicine components combination  Natural products  Atractylodes macrocephala polysaccharide  Geniposide  Chlorogenic acid  Intestinal mucosa barrier  Tight junction
基金项目:国家自然科学基金项目(81370094,81673750);上海市科委“浦江人才”计划项目(17PJ1408900),上海市“杏林新星”计划项目(ZY3-RCPY-2-2010)
作者单位
冷静1,2,田华捷1,2,黄甫1,2,冯琴1,2,赵瑜1,2,胡义扬1,3,4,彭景华1,2,3 1 上海中医药大学附属曙光医院上海201203 2 上海中医药大学肝病研究所上海201203 3 上海市中医临床重点实验室上海201203 4 上海中医药大学附属医院临床药理研究所上海201203 
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中文摘要:
      目的:观察中药组分复方BZL方(白术多糖、栀子苷、绿原酸)对高脂饮食诱导的小鼠脂肪性肝炎(NASH)及其肠黏膜损伤的作用。方法:C57BL/6J雄性小鼠27只,随机分为正常、模型和BZL方组,每组9只。除正常组外,其余各组均采用高脂饮食诱导NASH,第14周开始,BZL方组灌胃BZL方,其余各组灌胃等量饮用水,至17周末处死取材。观察各组小鼠体质量、进食量、肝体比,肝组织病理(HE和天狼猩红染色,SAF评分),检测血浆ALT、GGT活性、LPS水平,肝组织TG含量;肝组织胶原I型和IV型、内毒素受体CD14、TLR1、TLR2、TLR4、TLR9及炎性反应因子IL-1β、TNF-α mRNA表达(real-time PCR法);肠组织病理(HE染色);大肠组织紧密连接zo-1、occludin、claudin mRNA表达(real-time PCR法)及蛋白表达(免疫荧光染色)。结果:高脂饮食诱导NASH模型中,模型组血浆ALT、GGT、LPS及肝组织TG含量较正常组显著升高(P<0.01);肝组织可见脂肪沉积、炎性反应、气球样变及纤维化,SAF评分显著高于正常组,肝组织胶原ColI、ColIV mRNA显著升高,CD14、TLR1、TLR2、TLR4、TLR9及IL-1β、TNF-α mRNA表达升高(P<0.05,P<0.01);肠组织病理变化光镜下不明显,大肠组织紧密连接zo-1、occludin、claudin mRNA表达降低(P<0.01),zo-1、occludin免疫荧光阳性染色减少。BZL方可降低血浆ALT、GGT、LPS、肝组织TG含量(P<0.05,P<0.01),改善SAF评分,降低肝组织胶原ColI、ColIV mRNA表达(P<0.01),降低CD14、TLR2、TLR4、TLR9及IL-1β、TNF-α mRNA表达(P<0.05,P<0.01),恢复大肠组织紧密连接zo-1、occludin mRNA表达(P<0.01),恢复zo-1、occludin免疫荧光阳性染色。结论:BZL方有效减轻高脂饮食诱导的小鼠NASH,该作用与其保护肠黏膜、抑制LPS肠渗漏及肝组织中TLRs及炎性反应因子密切相关。
English Summary:
      To observe the effects of Chinese Medicine BZL recipe(composed of Atractylodes macrocephala polysaccharide,chlorogenic acid,and geniposide)on non-alcoholic steatohepatitis(NASH)and intestinal mucosal injury induced by high-fat diet in mice. Methods:Twenty seven male C57BL/6J mice were randomly divided into the normal group,the model group,and BZL group,with 9 rats in each group. Except the normal group,the other groups were induced by NASH. From the 14th week,mice in BZL group were administrated with BZL recipe by gavage daily,and the other mice were administrated with same dose distilled water. Till the end of 17th week,the mice were killed for collection materials. The body weight,food intake,liver-to-body ratio,liver histopathology(HE and Sirius red staining,SAF score)were observed in each group. Plasma ALT,GGT activity,LPS level,liver tissue TG content,liver tissue collagen type I and type IV,endotoxin receptor CD14,TLR1,TLR2,TLR4,TLR9 and inflammatory factors IL-1β,TNF-α mRNA expression(real-time PCR),intestinal histopathology(HE staining),large intestine tissue tight junction zo-1,occludin,claudin mRNA expression(real-time PCR)and protein expression(immunofluorescence staining)were detected.Results:In the high-fat diet induced NASH model,plasma ALT,GGT,LPS and liver tissue TG levels in the model group were significantly higher than those in the normal group(P<0.01); liver tissue showed fat deposition,inflammation,balloon-like changes and fibrosis. SAF score was significantly higher than the normal group. Liver tissue collagen ColI,ColIV mRNA increased significantly,and CD14,TLR1,TLR2,TLR4,TLR9 and IL-1β,TNF-α mRNA expression increased(P<0.05,P<0.01); The pathological changes of intestinal tissue were not obvious under light microscope. The expression of zo-1,occludin and claudin mRNA in the large intestine was decreased(P<0.01),and the immunofluorescence staining of zo-1 and occludin was decreased. BZL recipe can reduce plasma ALT,GGT,LPS,liver tissue TG content(P<0.05,P<0.01),improve SAF score,reduce liver tissue collagen ColI,ColIV mRNA expression(P<0.01),reduce CD14,TLR2,TLR4,TLR9,IL-1β and TNF-α mRNA expression(P<0.05,P<0.01),restored the expression of zo-1 and occludin mRNA in the large intestine,and restored the immunofluorescence staining of zo-1 and occludin. Conclusion:BZL recipe can effectively alleviate NASH induced by high-fat diet,which is closely related to the protection of intestinal mucosa,inhibition of LPS intestinal leakage and TLRs and inflammatory factors in liver tissue.
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