摘要
Objective To explore the therapeutic effect and mechanism of pachymic acid(PA)on Helicobacter py-lori(Hp)-associated gastritis in rats.Methods A rat model of Hp-associated gastritis was established;all rats were separated into control group(CT group),model group(group M),PA low-dose group(PA L group),PA high-dose group(PA H group),and PA H+phosphatidylinositol 3-kinase(PI3K)activator(740 Y-P)group;the gastric mucosal injury index(UI)of rats in each group was evaluated,transmission electron microscopy was applied to observe the morphology of gastric mucosal cells.HE staining was applied to evaluate the pathological characteristics of gastric mucosa.ELISA was applied to detect the levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),IL-10,induc-ible nitric oxide synthase(iNOS),and superoxide dismutase(SOD)in gastric tissue.Western blot method was applied to detect the expression of PI3K,phosphorylated PI3K(p-PI3K),protein kinase B(AKT),p-AKT,nuclear factor(NF)-κB p65,and p-NF-κB p65 proteins.Results Compared with the CT group,the gastric mucosa erosion,epithelial ede-ma,congestion,and severe ulcers were observed in the group M,with epithelial cell pyknosis and inflammatory cell in-filtration,the UI,IL-6,TNF-α,iNOS,and the expression levels of p-PI3K/PI3K,p-AKT/AKT,p-NF-κB p65/NF-κB p65 proteins increased,the levels of IL-10 and SOD decreased(P<0.05);compared with group M,the gastric mucosal damage and inflammatory cell infiltration in the PA L and PA H groups were improved,the UI,IL-6,TNF-α,iNOS by the host animal and the expression of p-PI3K/PI3K,p-AKT/AKT,p-NF-κB p65/NF-κB p65 proteins all decreased,the level of IL-10 and SOD was increased(P<0.05);compared with the PA H group,the pathological damage of the gastric mucosa in the PA H+740 Y-P group was aggravated,with epithelial cell pyknosis.The UI,IL-6,TNF-α,iNOS,and the expression of p-PI3K/PI3K,p-AKT/AKT,p-NF-κB p65/NF-κB p65 proteins increased,the levels of IL-10 and SOD decreased(P<0.05).Conclusions PA might facilitate the treatment of Hp-associated gastritis in rats by inhibiting the PI3K/AKT/NF-κB signaling pathway.
摘要
Aim To explore the mechanism of Polygonum capitatum(PC)in the treatment of Helicobacter Pylori associated gastritis(HAG). Methods The databases were used to identify the target of PC active compounds and HAG-related genes,and the intersection was taken to obtain the potential targets of PC treatment of HAG. The interaction network diagram of “drug-active compound-target-disease” and the protein-protein interaction(PPI)network of potential target protein interaction in HAG treated by PC were constructed by software Cytoscape 3.6.0. The important nodes in the network were screened by several topological indexes,and the GO and KEGG enrichment were analyzed by STRING database to obtain the potential signaling pathway of PC in the treatment of HAG. The binding ability of PC active components with key target proteins was observed by molecular docking method. On this basis,the related targets of PC in the treatment of HAG were verified in vivo and in vitro experiments. Results The PC active compounds and targets were identified through the database,and the “drug-active compound-target-disease” network diagram and the PPI network of potential target proteins were constructed. Combined with several topological indexes,the PPI network of potential target-protein interaction was analyzed,and 52 hub genes were screened. Further bioinformatics analysis and high-throughput sequencing revealed that PC exerted an effect on HAG through the Akt/NF-κB/NLRP3 pathway. Based on this,it was found that PC could reduce IL-18 and IL-1β in HAG GES-1 cells and HAG SD rats,up-regulate Akt and its phosphorylation level and reduce NF-κB expression,inhibit the activation of NLRP3 inflammatory body,so as to improve HAG inflammatory response. Conclusions PC could exert a therapeutic effect on HAG by activating Akt and its phosphorylation level,and inhibiting the expression of NF-κB and NLRP3 inflammasome related factors. This study provides a theoretical basis for explaining the mechanism of PC in the treatment of HAG.
摘要
Objective The activation of NF-kappa B (NF-κB) signaling pathway plays an important role in the development of helicobacter pylori associated gastritis (HAG). The article aimed to investigate the effects of polygonum capitatum on the treatment of HAG in NF-κB signaling pathway and observe whether the regulation of NF-κB acetylation by silent information regulator 1 (SIRT1) affects the therapeutic effects of HAG. Methods The immortalized human gastric epithelial cells (GES-1) were cultured and the H.pylori stand- ard strain ATCC700392 was used for the replication of HAG cell model by 100∶1. The cells were divided into model group,drug group and normal control group. Cells were treated with 80 μg/mL in drug group,H.pylori and GES-1 were cultured together in model group and untreated GES-1 cells were taken as control group. Real time PCR was used to detect the mRNA levels of SIRT1,NF-κB/p65 and TNF-α. Western blotting was used to detect the expression levels of SIRT1,NF-κB/p65 and its acetylated protein in the total protein,as well as the expression levels of SIRT1 and NF-κB/p65 in cytoplasm and nuclear protein. Results At 12 h after the infection of H. pylori,the level of TNF-α in the supernatant was higher than that in the normal control group(P<0.05). The expression of SIRT1 de-creased in the cytoplasm of model group,while the expression levels of NF-κB/p65,acetyl-NF-κB p65(Lys310) and TNF-α in the nu-cleus increased (P<0.05). But after the treatment of polygonum capitatum,the expression of SIRT1 in the nucleus increased(P<0.05) while the expression of NF-κB/p65,acetyl-NF-κB p65(Lys310) and TNF-α decreased (P<0.05). Conclusion Polygonum capita-tum can activate the SIRT1 in the nucleus,which makes activated NF-κB/p65 in the nucleus carry out deacetylation modification in or-der to antagonize the cell damage induced by H.pylori.