基于TLR4/MYD88/NF-κB信号通路和肠道菌群探讨罗乐胃蜜丸对慢性非萎缩性胃炎大鼠的作用及机制OA

Objective:This paper aims to investigate the therapeutic effects and potential mechanism of Luolewei Honey Pills(罗乐胃蜜丸)on chronic non-atrophic gastritis(CNAG)rats.Methods:The CNAG rat model was estab-lished by a multi-factor combined stimulation method.The successfully modeled rats were randomly divided into a model control group,Luolewei Honey Pill groups(0.2,0.4,and 0.8 g/kg),and a Weisu Granule group(4.7 g/kg),and a normal control group was set.Each group received intragastric administration for four weeks.The general state and body mass changes of the rats were observed.The gastric juice pH value and pepsin activity were measured.The levels of in-terleukin-6(IL-6),interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),gastrin(GAS),and motilin(MTL)in serum were determined by enzyme-linked immunosorbent assay(ELISA).Histopathological changes of the gastric muco-sa were observed by hematoxylin-eosin(HE)staining.The protein expressions of toll-like receptor 4(TLR4),myeloid differentiation primary response protein 88(MYD88),and nuclear factor kappa-B p65(NF-κB p65)in gastric tissue were detected by Western blot.Changes in fecal intestinal microbiota structure were analyzed by 16S rRNA high-through-put sequencing,and the correlations between microbiota and inflammatory factors,as well as gastrointestinal hormones,were evaluated.In addition,normal mice and model mice with gastrointestinal motility disorder induced by atropine were used to observe the effects of Luolewei Honey Pills on gastric emptying rate and small intestinal propulsion rate.Results:Compared with those in the normal control group,the rats in the model control group showed significantly slower body weight gain and significantly decreased pepsin activity(P<0.01).The gastric juice pH value and the levels of IL-6,IL-1β,TNF-α,and GAS in serum were increased(P<0.01),and the MTL content was significantly decreased(P<0.01).Obvious pathological damage was observed in the gastric mucosa.Protein expressions of TLR4,MYD88,and NF-κB p65 were significantly up-regulated(P<0.01).Intestinal microbial diversity was altered,characterized by a decreased rela-tive abundance of beneficial bacteria such as Bacteroidetes and an increased relative abundance of harmful bacteria such as Proteobacteria.Compared with those in the model control group,the body weight and pepsin activity of rats in the Lu-olewei Honey Pill groups were significantly increased(P<0.01),while gastric juice pH value and the contents of IL-6,IL-1β,TNF-α,and GAS in serum were decreased(P<0.05 or P<0.01).The MTL content was significantly increased(P<0.01).In addition,the pathological damage of the gastric mucosa was alleviated,and the protein expressions of TLR4,MYD88,and NF-κB p65 were down-regulated(P<0.05 or P<0.01).Luolewei Honey Pills increased the relative abundance of beneficial bacteria like Bacteroidetes and decreased the relative abundance of harmful bacteria like Pro-teobacteria.The abundance of protective microbiota(e.g.,Bacteroidetes)was negatively correlated with inflammatory factors and GAS and positively correlated with MTL,while the abundance of harmful microbiota(e.g.,Proteobacteria)showed the opposite correlations.Luolewei Honey Pills significantly increased the gastric emptying rate and small intesti-nal propulsion rate in the model mice with gastrointestinal motility disorder induced by atropine(P<0.05 or P<0.01).Conclusion:Luolewei Honey Pills can effectively ameliorate gastric mucosal inflammatory injury in CNAG rats.The mechanism may be related to inhibiting the activation of the TLR4/MYD88/NF-κB signaling pathway,alleviating inflam-matory responses,regulating gastrointestinal hormones,and remodeling the intestinal microbiota structure.