[目的]探究桦褐孔菌乙醇提取物(ethanol extract of Inonotus obliquus,EEIO)对脂多糖(lipopolysaccharides,LPS)诱导肠损伤的保护作用及分子机制.[方法]利用LPS诱导的小鼠小肠上皮细胞和C57BL/6小鼠,检测EEIO处理后LDH和NO水平;RT-qPCR和ELISA方法检测EEIO对炎症因子IL-1β和IL-18表达的影响;RT-qPCR和WB方法检测EEIO对MAPK和焦亡信号通路的影响.[结果]体内外结果一致表明,EEIO可有效缓解LPS导致的LDH和NO水平升高;EEIO处理后,IL-1β和IL-18表达明显降低.EEIO抑制MAPK(ERK、P38、JNK)信号通路,显著降低p-ERK/ERK、p-p38/p38和p-JNK/JNK的水平.同时,焦亡途径被抑制,ASC、Caspase-1、GSDMD和NLRP3水平显著降低.阳性药物白藜芦醇(resveratrol,RES)对各项检测指标的影响与EEIO相似,但效果不如EEIO.[结论]EEIO通过调控MAPK信号通路,抑制焦亡途径进而降低炎性因子的表达,缓解LPS诱导肠损伤.
Alleviating Roles and Mechanisms of Ethanol Extract from Inonotus obliquus to Intestinal Injury Induced by Lipopolysaccharide
[Objective]To investigate the protective effect and molecular mechanism of ethanol extract of Inonotus obliquus(EEIO)on lipopolysaccharide(LPS)-induced intestinal injury.[Method]Using LPS to induce mouse small intestinal epithelial cells and C57BL/6 mice,LDH and NO levels were measured after EEIO treatment.RT qPCR and ELISA methods were to detect the effects of EEIO on the expressions of inflammatory factor IL-1 β and IL-18.RT-qPCR and WB methods were used to detect the effects of EEIO on MAPK and pyroptosis signaling pathways.[Result]Both in vivo and in vitro results showed that EEIO effectively alleviated the increase in LDH and NO levels induced by LPS.After EEIO processing,the expressions of IL-1β and IL-18 were significantly reduced.EEIO inhibited the MAPK(ERK,P38 and JNK)signaling pathway,significantly reducing the levels of p-ERK/ERK,p-p38/p38 and p-JNK/JNK.Meanwhile,the pyroptosis pathway was inhibited,and the levels of ASC,Caspase-1,GSDMD and NLRP3 significantly reduced.The positive drug resveratrol(RES)had a similar effect on various detection indicators as EEIO,but its effect was not as good as EEIO.[Conclusion]EEIO regulates the MAPK signaling pathway,inhibits the pyroptosis pathway,and reduces the expression of inflammatory factors,thus alleviating LPS-induced intestinal injury.
ethanol extract of Inonotus obliquuslipopolysaccharidesintestinal injuryinflammationpyroptosis
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