目的:脓毒症发病率高,发病率,和死亡率,是对人类安全的巨大威胁。肠道健康在脓毒症的发展中起着重要作用。齐黄方(QHF)含有黄芪,大黄,zhishi,还有白术.用于治疗气虚和正虚综合征。本研究旨在探讨QHF是否通过NLRP3炎性囊泡介导的细胞局灶性死亡改善小鼠肠屏障功能和微生物。
方法:采用无特异性病原体(SPF)级C57BL/6小鼠盲肠结扎穿孔(CLP)法,中等,和高剂量的黄芪配方或益生菌4周。术后24小时,QHF缓解脓毒症肠功能障碍和恢复肠道微生态的作用机制,从而减轻肠道损伤,通过病理观察进行评估,免疫组织化学,西方印迹,ELISA,和16SrDNA高通量测序。
结果:不同剂量的QHF和益生菌均有不同程度的改善小鼠肠道损伤和减少结肠细胞凋亡(P<0.05)。同时,不同剂量的QHF和益生菌能够降低血清IL-6,IL-1β,和TNF-α(P<0.05);下调NLRP3,caspase-1和caspase-11的蛋白表达(P<0.05);上调zonulaoccluden-1(ZO-1)和occludin的蛋白表达(P<0.05),改善小鼠肠道屏障功能。此外,QHF降低了有害细菌的相对丰度(Firmicutes,Muribaculaceae,弯曲杆菌,螺杆菌,和Alistipes),并增加了有益菌(拟杆菌和放线菌)的相对丰度(P<0.05)。
结论:QHF通过NLRP3炎性体介导的细胞焦亡改善小鼠肠屏障功能和肠道微生物学。
OBJECTIVE: Sepsis has a high incidence, morbidity, and mortality rate and is a great threat to human safety. Gut health plays an important role in sepsis development. Qi Huang Fang (QHF) contains astragalus, rhubarb, zhishi, and atractylodes. It is used to treat syndromes of obstructive qi and deficiency of righteousness. This study aimed to investigate whether QHF improves intestinal barrier function and microorganisms in mice through NLRP3 inflammatory vesicle-mediated cellular focal death.
METHODS: A mouse model of sepsis was constructed by cecal ligation and puncture (CLP) of specific pathogen-free (SPF)-grade C57BL/6 mice after continuous gavage of low, medium, and high doses of astragalus formula or probiotics for 4 weeks. Twenty-four hours postoperatively, the mechanism of action of QHF in alleviating septic intestinal dysfunction and restoring intestinal microecology, thereby alleviating intestinal injury, was evaluated by pathological observation, immunohistochemistry, western blotting, ELISA, and 16S rDNA high-throughput sequencing.
RESULTS: Different doses of QHF and probiotics ameliorated intestinal injury and reduced colonic apoptosis in mice to varying degrees (P < 0.05). Meanwhile, different doses of QHF and probiotics were able to reduce the serum levels of IL-6, IL-1β, and TNF-α (P < 0.05); down-regulate the protein expression of NLRP3, caspase-1, and caspase-11 (P < 0.05); and up-regulate the protein expression of zonula occluden-1 (ZO-1) and occludin (P < 0.05), which improved the intestinal barrier function in mice. In addition, QHF decreased the relative abundance of harmful bacteria (Firmicutes, Muribaculaceae, Campilobacterota, Helicobacter, and Alistipes) and increased the relative abundance of beneficial bacteria (Bacteroidetes and Actinobacteria) (P < 0.05).
CONCLUSIONS: QHF improves intestinal barrier function and gut microbiology in mice via NLRP3 inflammasome-mediated cellular pyroptosis.