机构地区:[1]首都医科大学附属北京朝阳医院中医科,北京100020
出 处:《山西医科大学学报》2025年第2期142-150,共9页Journal of Shanxi Medical University
基 金:北京市级中西医协同“旗舰”科室建设项目;朝阳区区级中医重点专科培育类项目;北京市医管局培育计划项目(PZ2022003)。
摘 要:目的 探索和肝汤对功能性消化不良(functional dyspepsia,FD)的作用机制是否与肠道菌群以及Toll样受体4(Tolllike receptor 4,TLR4)信号通路有关。方法 18只6周龄SD大鼠随机分为正常组、模型组以及和肝汤组,每组6只。模型组与和肝汤组大鼠以夹尾应激法建立FD模型,造模后和肝汤组予0.918 g生药/100 g体质量的和肝汤灌胃7 d,干预结束后取十二指肠组织和盲肠内容物,荧光定量PCR检测TLR4基因转录水平,蛋白印迹法检测TLR4蛋白表达水平,免疫荧光和免疫组化法观察TLR4的表达位置,16S rRNA和液相色谱-质谱联用分析盲肠内容物中菌群和代谢物情况。结果 与正常组比较,模型组TLR4蛋白表达水平显著增加(P<0.01);与模型组比较,和肝汤组TLR4表达下调(P<0.05)。免疫荧光和免疫组化显示,TLR4在各组十二指肠固有层和上皮层均存在表达。模型组相比正常组阳性着色增强(P<0.01),和肝汤组相比模型组阳性着色减弱(P<0.01);但各组TLR4基因转录水平无显著差异(P>0.05)。与正常组比较,模型组盲肠内容物中疣微菌门丰度增加(P<0.05),而厚壁菌门丰度减低(P<0.05);与模型组比较,和肝汤组拟杆菌门丰度提升(P<0.05),疣微菌门丰富下调(P<0.05);各组间n-γ-谷氨酰谷氨酰胺等代谢物含量存在一定差异(P<0.05)。结论 FD大鼠十二指肠TLR4表达上调,同时肠道菌群和代谢产物的结构存在异常改变。和肝汤可通过调节肠道菌群和代谢产物的结构,下调TLR4在十二指肠的表达改善FD。Objective To explore the effect of Hegantang granules on functional dyspepsia(FD)and its mechanism based on gut mi-crobiota and the Toll-like receptor 4(TLR4)signaling pathway.Methods Eighteen 6-week-old SD rats were randomly divided into normal group,model group,and Hegantang group,with 6 rats in each group.FD model was established using tail-clamping stress in model group and Hegantang group.After modeling,the rats in Hegantang group were given 0.918 g crude drug/100 g body weight by gavage for 7 d.After the intervention,duodenum tissues and cecal contents were collected.The transcription level of TLR4 gene was detected by quantitative polymerase chain reaction(qPCR).TLR4 protein expression was assessed by Western blotting.And the ex-pression location of TLR4 was observed using immunofluorescence(IF)and immunohistochemistry(IHC).Additionally,16S rRNA and liquid chromatograph-mass spectrometer(LC-MS)were used to analyze the gut microbiota and metabolites in the cecal contents.Results Compared to normal group,TLR4 protein expression was significantly increased in model group(P<0.01).In comparison with model group,TLR4 expression was downregulated in Hegantang group(P<0.05).IF and IHC results revealed that TLR4 ex-pressed in both lamina propria and epithelium of the duodenum in all groups.Compared with normal group,positive staining was en-hanced in model group(P<0.01),while it was reduced in Hegantang group compared with model group(P<0.01).However,there was no significant difference in TLR4 gene transcription level among the groups(P>0.05).Compared with normal group,the abundance of Verrucomicrobiota was increased in model group(P<0.05),while the abundance of Firmicutes was decreased(P<0.05).Compared with model group,the abundance of Bacteroidota was increased in Hegantang group(P<0.05),and the abundance of Verrucomicro-biota was downregulated(P<0.05).Additionally,there were differences in the levels of metabolites such as n-γ-glutamylglutamine among the groups(P<0.05).Conclusion The expression of TLR4 in the d
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