湖南中医药大学学报2025，Vol.45Issue(11)：2044-2053,10.DOI:10.3969/j.issn.1674-070X.2025.11.004
基于TLR4/MyD88/NF-κB通路与肠道菌群探究仲叶止泻合剂对脾虚泄泻小鼠的作用机制
Exploring the mechanism of action of Zhongye Zhixie Mixture on mice with spleen-deficiency diarrhea based on the TLR4/MyD88/NF-κB pathway and gut microbiota
摘要
Abstract
Objective To investigate the effects and mechanism of Zhongye Zhixie Mixture(ZYZXM)on mice with spleen-deficiency diarrhea based on the Toll-like receptor 4/myeloid differentiation primary response protein 88/nuclear factor-kappa B(TLR4/MyD88/NF-κB)pathway and gut microbiota.Methods Gavage administration of rhubarb decoction was performed to establish a spleen-deficiency diarrhea model.Sixty male KM mice were randomly divided into six groups,including a blank group,a model group,a positive drug group,and high-,medium-,and low-dose groups of ZYZXM,with 10 mice in each group.After successful model establishment,each drug administration group received corresponding medication via gavage for 7 days.The body weight,diarrhea condition,and spleen index of the mice were compared before and after treatment.Hematoxylin-eosin(HE)staining was used to observe the morphological changes in the colon tissues of mice in each group.16S rDNA high-throughput sequencing was employed to check alterations in the gut microbiota structure of mice in each group.Enzyme-linked immunosorbent assay(ELISA)was utilized to measure the levels of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in the serum of mice in each group.Western blot analysis was performed to examine the protein expression levels of TLR4 and MyD88 in the colon tissues of mice in each group.Immunofluorescence was used to check the nuclear translocation of nuclear factor-kappa B(NF-κB)p-p65 protein in the colon tissues of mice in each group.Results Compared with the blank group,the mice in the model group exhibited decreases in body weight and spleen index(P＜0.05),while increases were observed in fecal water content,diarrhea index,serum levels of IL-6 and TNF-α,protein expression levels of TLR4 and MyD88 in colon tissues,and relative fluorescence intensity of nuclear NF-κB p-p65 in colon tissues(P＜0.05).In comparison with the model group,the mice in the positive drug group and different-dose groups of ZYZXM showed increases in body weight and spleen index(P＜0.05),along with decreases in fecal water content,diarrhea index,serum levels of IL-6 and TNF-α,protein expression levels of TLR4 and MyD88 in colon tissues,and relative fluorescence intensity of nuclear NF-κB p-p65 in colon tissues(P＜0.05).When compared with the positive drug group and the high-and medium-dose groups of ZYZXM,the low-dose group of ZYZXM demonstrated decreases in fecal water content,diarrhea index,serum levels of IL-6 and TNF-α,protein expression levels of TLR4 and MyD88 in colon tissues,and relative fluorescence intensity of nuclear NF-κB p-p65 in colon tissues(P＜0.05).The results of 16S rDNA sequencing indicated that ZYZXM could regulate the gut microbiota structure in mice,restore α and β diversity,increase the relative abundance of beneficial bacteria,and decrease the relative abundance of harmful bacteria.Conclusion ZYZXM can alleviate diarrhea symptoms in mice with spleen-deficiency diarrhea and improve pathological damage in colon tissues.This effect may be achieved by down-regulating the expression of proteins in the TLR4/MyD88/NF-κB signaling pathway and improving the gut microbiota.关键词
腹泻/仲叶止泻合剂/脾虚泄泻/绿原酸/TLR4/MyD88/NF-κB通路/肠道菌群Key words
diarrhea/Zhongye Zhixie mixture/spleen-deficiency diarrhea/chlorogenic acid/TLR4/MyD88/NF-κB/gut microbiota分类
医药卫生引用本文复制引用
朱媛媛,潘龙,韩曼,宋延平..基于TLR4/MyD88/NF-κB通路与肠道菌群探究仲叶止泻合剂对脾虚泄泻小鼠的作用机制[J].湖南中医药大学学报,2025,45(11):2044-2053,10.基金项目
陕西省中医药管理局项目(ZYJXG-Y23009). （ZYJXG-Y23009）
