海南医科大学学报2025，Vol.31Issue(10)：721-731,11.DOI:10.13210/j.cnki.jhmu.20240906.001
海南萝芙木果胶多糖通过MAPK/NF-κB信号通路调控小鼠溃疡性结肠炎的焦亡机制研究
The pyroptosis mechanism of pectic polysaccharides of Rauvolfia erticillata(Lour.)Baill.var.hainanensis Tsiang in regulating ulcerative colitis mice through MAPK/NF-κB signaling pathway
摘要
Abstract
Objective:To investigate the therapeutic efficacy of pectic polysaccharides(PPs)on dextran sulfate sodium salt(DSS)induced ulcerative colitis in mice.The therapeutic effect of ulcerative colitis(UC)and its role in mediating mitogen-activated protein kinase(MAPK)/nuclear factor kappa-B(NF-κB),and effect of NF-κB/nucleotide-binding oligomeriza-tion domain-like receptorprotein 3(NLRP3)signaling pathway on pyroptosis in colonic epithelial cells was also identified.Meth-ods:The study utilized a total of 24 male C57 mice,aged between 6-8 weeks,which were randomly assigned into three groups:control group,model group,and PPs treatment group.Each group consisted of 8 mice.In addition to the control group,the other two groups were given 5%dextran sulfate sodium salt(DSS)to drink freely for 7 days to prepare UC mouse model.After success-ful modeling,the mice in the PPs treatment group continued to receive gavage for 7 days.The changes in body weight,presence of blood in stool,and disease activity index(DAI)of the mice were recorded,and the whole colon was separated and measured.The pathological changes of colon were observed by hematoxylin-eosin(HE)staining.The expressions of caspase-1,IL-1β and TNF-α were detected by immunohistochemistry,while pyroptosis(caspase-1,IL-18,IL-1β,NLRP3,Cleaved GSDMD)and the protein expression of MAPK/NF-κB pathway(p38,p-p38,p65,p-p65,JNK,p-JNK,ERK,p-ERK)in colon tissues was analyzed using Western blotting.The morphology of pyroptosis was observed by transmission electron microscope.Dual luciferase reporter gene assay and ChIP assay were used to detect NF-κB p65 binding to NLRP3 promoter.Results:Compared with the model group,treatment with PPs attenuated weight loss in mice and inhibited colon shortening while alleviating colon pathological injury;DAI score was lower than that of DSS model group(P<0.05).The protein expressions of NLRP3,caspase-1,IL-1β,IL-18,TNF-α,cleaved GSDMD in colon epithelial cells were significantly decreased(P<0.01),and the expressions of p38,P-P38,p65,P-P65,JNK,P-JNK,ERK,P-ERK proteins in colon tissues were significantly decreased(P<0.05).The morphol-ogy of pyroptosis in some cells was observed by transmission electron microscope.Dual luciferase reporter gene assay and ChIP as-say confirmed that NF-κB p65 gene targeted the promoter region of NLRP3 gene,and PPs promoted NLRP3 transcription by en-hancing it.Conclusion:PPs may exert an inhibitory effect on cell pyroptosis through the modulation of the MAPK/NF-κB signal-ing pathway,subsequently attenuating colonic inflammation in DSS-induced mice models.The transcriptional binding of NF-κB to the NLRP3 promoter may be represents a pivotal mechanism that influences the pyroptosis of colorectal cells.关键词
溃疡性结肠炎/果胶多糖/海南萝芙木/MAPK信号通路/核因子-κB/细胞焦亡Key words
Ulcerative colitis/Pectic polysaccharides/Rauvolfia erticillata(Lour.)Baill.var.hainanensis Tsiang/MAPK signaling pathway/Nuclear factor-κB/Pyroptosis分类
中医学引用本文复制引用
王柳丹,陈旭永,吴海东,苗新普..海南萝芙木果胶多糖通过MAPK/NF-κB信号通路调控小鼠溃疡性结肠炎的焦亡机制研究[J].海南医科大学学报,2025,31(10):721-731,11.基金项目
This study was supported by Natural Science Foundation of Hainan Province(821MS126) （821MS126）
Hainan Provincial Department of Education Key Project(Hnky2021ZD-13) （Hnky2021ZD-13）
Hainan Province Graduate Innovation Research Project(HYYB2023A25) （HYYB2023A25）
Hainan Province Key R&D Program Project(ZDYF2022SHFZ099) 海南省自然科学基金(821MS126) （ZDYF2022SHFZ099）
海南省教育厅重点课题(Hnky2021ZD-13) （Hnky2021ZD-13）
海南省研究生创新科研课题(HYYB2023A25) （HYYB2023A25）
海南省重点研发计划项目(ZDYF2022SHFZ099) （ZDYF2022SHFZ099）
