基础医学与临床 ›› 2017, Vol. 37 ›› Issue (9): 1263-1269.

• 研究论文 • 上一篇    下一篇

合生元调整肠道微生态治疗大鼠NASH及其对TLR4的影响

蔡洁毅,罗丹萍,张扬,袁瑜,何兴祥   

  1. 广东药学院附属第一医院
  • 收稿日期:2016-02-22 修回日期:2016-11-04 出版日期:2017-09-05 发布日期:2017-08-28
  • 通讯作者: 何兴祥 E-mail:13724887928@qq.com
  • 基金资助:
    广东省医学科研基金资助项目

Synbiotics adjusting the intestinal microecologe to treat rats NASH and its effect on TLR4

  • Received:2016-02-22 Revised:2016-11-04 Online:2017-09-05 Published:2017-08-28

摘要: 目的 探讨肠道菌群改变在非酒精性脂肪性肝炎(NASH)形成过程中的作用,进而探索通过合生元干预治疗NASH。方法 以饲喂高脂饲料建非酒精性脂肪性肝炎(NASH)模型,动态观察大鼠自主活动次数;全自动生化分析仪检测大鼠血清的甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白(HDL)、低密度脂蛋白(LDL)、空腹血糖(FBS)及空腹胰岛素(FINS)的含量;用16S rRNA荧光定量PCR检测大鼠肠道主要菌群;常规HE染色观察肝组织病理学,计算NAFLD活动度积分(NAS);用酶联免疫吸附试验和免疫组织化学法检测TLR4表达。在NASH建模第4,8及12周分别取10只大鼠,添加合生元配方饲养2周(BIO组),继续观察上述指标。结果 1)随着高脂饮食喂养时间的延长,肝细胞脂肪变程度明显加重,NAFLD评分显著增高(P<0.01)。2)合生元干预2周后大鼠自主活动次数显著上调、血清学指标TG、TC、LDL、FBS和FINS水平显著下调(P<0.05)。3)合生元干预2周后,可使双歧杆菌和乳酸杆菌的数量显著上调,肠球菌的数量显著下调。4)NASH模型组TLR4的表达逐渐增高(P<0.05),经合生元干预2周后可显著下调大鼠TLR4的表达(P<0.05)。结论 肠道微生态改变与NASH的发生发展密切相关,合生元通过调整肠道微生态改善NASH鼠的生活质量与生化指标,其机制可能与TLR4蛋白水平回调有关。

关键词: 非酒精性脂肪性肝炎, 肠道微生态, 益生菌, 益生元, 合生元

Abstract: Objective To observe the change of intestinal flora on the process of nonalcoholic steatohepatitis(NASH),and to explore the synbiotics therapeutic effect on NASH. Methods Rats were administrated with high fat diet to establish NASH model.In the process of NASH rats modeling,the level of triglyceride (TG), total cholesterol (TC), high-density lipoprotein (HDL), low density lipoprotein (LDL), fasting blood sugar (FBS) and fasting insulin (FINS) was tested dynamically by automatic biochemical analyzer.The changes of main intestinal flora were detected by 16 S rRNA fluorescence quantitative polymerase chain reaction.NAFLD activity score was calculated. HE staining was used to observe the hepaticpathological changes and the TLR4 expression was detected by using enzyme-linked immunosorbent assay and immunohistochemical method. Until the 4th,8th,10th week in the process of NASH modeling, 10 rats were feeded with Synbiotics for 2 weeks, and all of above indicators were tested and observed. Results 1) With the extension of a high-fat diet feeding time, the degree of hepatocyte steatosis obviously was increased.NAFLD score was significantly heightened(P < 0.01).2)Number of independent activities of rats was significantly increased, the serological level of TG, TC, LDL, FBS and FINS were lower significantly after intervention with synbiotics for 2 weeks(P < 0.05). 3) Synbiotics intervention for two weeks could significantly increase the amount of bifidobacterium and lactobacillus and decrease the amount of enterococcus significantly(P < 0.05). 4) The expression of TLR4 was gradually increased in the process of NASH rats modeling(P < 0.05),but decreased after 2 weeks of the synbiotics-intervention (P < 0.05). Conclusions Intestinal microecology change is closely related to the development of NASH,therefor, syinbiotics could improve the quality of life and biochemical indicators of NASH rats through adjusting intestinal microecology and the expression level of TLR4 protein might been involved.

Key words: nonalcoholic steatohepatitis(NASH), intestinal microflora, probiotics, prebiotics, synbiotics