背景:黄芪(AM)及其有效成分主要用于抗炎,抗病毒,抗氧化剂,免疫调节,心血管和神经系统保护,抗癌,抗肿瘤等。
目的:探讨黄芪提取物改善溃疡性结肠炎(UC)的药理机制和生物学过程。
方法:建立葡聚糖硫酸钠(DSS)诱导的C57BL/6小鼠UC模型,并用黄芪提取物或salazo磺胺吡啶(SASP)治疗小鼠。使用DSS诱导的小鼠和人来源的结肠上皮细胞系来揭示UC的炎症环境。用黄芪提取物治疗后,实时定量PCR(qRT-PCR)检测细胞中磷脂酶C-β2(PLCB2)的表达,细胞计数试剂盒8(CCK-8)检测细胞增殖活性。最后,ELISA法检测细胞培养上清液中与焦亡相关的炎症因子水平。
结果:用黄芪提取物治疗UC小鼠可显著改善DAI评分和组织病理学损伤评分,降低了Eotaxin的水平,GCSF,KC,MCP-1,TNF-α,IL-6此外,黄芪提取物抑制核苷酸结合寡聚化片段样受体家族3(NLRP3)的表达,结肠组织中裂解的Caspase-1和GSDMD-N,降低血清和组织中炎症相关因子IL-1β和IL-18的水平。体外,黄芪提取物部分逆转了DSS诱导的CP-M030和NCM460中PLCB2表达的减少,促进了细胞增殖活性,并降低IL-1β和IL-18的水平。
结论:在DDS诱导的UC小鼠中,黄芪提取物通过PLCB2促进抑制结肠上皮细胞焦亡,改善溃疡性结肠炎。
BACKGROUND: Astragalus mongholicus Bunge (AM) and its active ingredients are mainly used for anti-inflammatory, antiviral, antioxidant, immune regulation, cardiovascular and nervous system protection, anti-cancer, anti-tumor and so on.
OBJECTIVE: To explore the Astragalus mongholicus Bunge extract pharmacological mechanisms and biology processes which improves ulcerative colitis (UC).
METHODS: Dextran sulfate sodium (DSS)-induced UC models in C57BL/6 mice were established, and the mice were treated with Astragalus mongholicus Bunge extract or salazosulfapyridine (SASP). DSS-induced mice- and human-derived colonic epithelial cell lines were used to reveal the inflammatory environment of UC. After treatment with Astragalus mongholicus Bunge extract, the expression of phospholipase C-β 2 (PLCB2) in the cells was detected by quantitative real-time PCR (qRT-PCR), and cell proliferative activity was detected by cell counting kit 8 (CCK-8) assay. Finally, the levels of pyroptosis-related inflammatory factors in cell culture supernatants was detected by ELISA.
RESULTS: Treatment of UC mice with Astragalus mongholicus Bunge extract do significantly improved DAI scores and histopathological damage scores, and decreased the levels of Eotaxin, GCSF, KC, MCP-1, TNF-α, and IL-6. Besides, Astragalus mongholicus Bunge extract inhibited the expression of nucleotide-binding oligomerization segment-like receptor family 3 (NLRP3), cleaved Caspase-1, and GSDMD-N in the colonic tissues, and reduced the levels of inflammation-related factors IL-1β and IL-18 in serum and tissues. In vitro, Astragalus mongholicus Bunge extract partially reversed the DSS-induced reduction of PLCB2 expression in CP-M030 and NCM460, promoted cell proliferative activity, and reduced the levels of IL-1β and IL-18.
CONCLUSIONS: In DDS-induced UC mice, Astragalus mongholicus Bunge extract improves ulcerative colitis by inhibiting colonic epithelial cell pyroptosis through PLCB2 promotion.