地五养肝方调控TLR4/NF-κB信号通路对肝癌模型肠道菌群调节作用  

Effect of Diwu Yanggan Decoction(地五养肝方)on Intestinal Flora in Liver Cancer Model Through Regulating TLR4/NF-κB Signal Pathway

作　　者：韩丹丹 胡珉芳 喻灿[1] HAN Dandan;HU Minfang;YU Can(Wuhan Hospital of Taditional Chinese Medicine,Wuhan 430000,Hubei,China)

机构地区：[1]武汉市中医医院,湖北武汉430000

出　　处：《辽宁中医药大学学报》2023年第11期68-72,共5页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：湖北省自然科学基金面上项目(2020CFB631);武汉市医学科研重点面上项目(WZ19A15)。

摘　　要：目的分析地五养肝方调控TLR4/NF-κB信号通路对肝癌模型肠道菌群及其菌群代谢功能的调节作用。方法选择24只SPF级大鼠随机分为对照组、模型组和观察组;模型组和观察组建立大鼠肝癌模型;末次给药后24 h时大鼠处死后取出肝脏组织,观察肝组织病理学变化;采用酶联免疫吸附法检测CA19-9、AFP、ALT水平;采用变性梯度凝胶电泳法(DGGE)绘制DGGE指纹图谱;利用气相色谱法检测大鼠粪便中短链脂肪酸水、D-乳酸水平;采用Western blotting法检测瘤组织中TLR4、NF-κB、GAPDH水平。结果对照组大鼠肝功能正常无受损,肝脏健全,模型组出现明显的癌细胞,且肝脏受损严重,观察组大鼠肝脏结构较为完整,肝脏组织炎性细胞减少,损伤程度减弱;观察组大鼠血清标志物CA19-9、AFP及ALT水平均明显低于模型组,且差异存在统计学意义(P<0.05);观察组大鼠大肠道菌群中双歧杆菌、乳酸杆菌、大肠埃希菌、肠球菌水平均明显低于模型组,且差异存在统计学意义(P<0.05);观察组大鼠肠道菌群DGGE指纹图谱的多样性指数、丰富度和均匀度明显高于模型组,且差异存在统计学意义(P<0.05);观察组大鼠TLR4蛋白和NF-κB蛋白表达水明显低于模型组,且差异存在统计学意义(P<0.05)。结论地五养肝方可有效调控TLR4/NF-κB信号通路,并改善肝癌模型肠道菌群及其菌群代谢功能。Objective To analyze the regulatory effect of Diwu Yanggan Decoction(地五养肝方)on TLR4/NF-κB signaling pathway on intestinal microflora and its metabolic function in hepatocellular carcinoma model.Methods Twenty-four SPF rats were randomly divided into control group,model group and observation group.Model group and observation group were used to establish rat hepatocellular carcinoma model.24 h after the last administration,the rats were sacrificed and the liver tissues were removed to observe the histopathological changes of the liver.The levels of CA19-9,AFP and ALT were detected by enzymelinked immunosorbent assay.DGGE fingerprints were obtained by denaturing gradient gel electrophoresis(DGGE).The levels of short-chain fatty acid water and D-lactic acid in feces of mice were determined by gas chromatography.TLR4,NF-κB and GAPDH levels in tumor tissues were detected by Western blotting.Results The liver function of the control group was normal without damage,and the liver was healthy.The model group showed obvious cancer cells,and the liver was seriously damaged.The liver structure of the observation group was relatively intact,and the inflammatory cells in the liver tissue were reduced,and the damage degree was weakened.The levels of serum markers CA19-9,AFP and ALT in observation group were significantly lower than those in model group,and the differences were statistically significant(P<0.05).The levels of Bifidobacteria,Lactobacillus,Escherichia coli and Enterococcus in the intestinal flora of rats in the observation group were significantly lower than those in the model group,and the differences were statistically significant(P<0.05).The diversity index,richness and evenness of DGGE fingerprints of intestinal flora in observation group were significantly higher than those in model group,and the differences were statistically significant(P<0.05).The expression levels of TLR4 protein and NF-κB protein in the observation group were significantly lower than those in the model group,and the differences

关 键 词：地五养肝方 TLR4/NF-κB信号通路 肝癌模型 肠道菌群 菌群代谢功能 

分 类 号：R285.5[医药卫生—中药学]