制附子总生物碱抗小鼠溃疡性结肠炎的作用及其机制研究

Anti-Ulcerative Colitis Effects and the Mechanisms of Total Alkaloids Extracted from Processed Aconitum carmichaelii Debx

目的本实验旨在初步探索制附子的总生物碱成分(Aconitum carmichaelii Debx.alkaloids,ACA)对溃疡性结肠炎(ulcerative colitis,UC)小鼠的潜在治疗作用及其相关作用机制。方法通过液质联用技术对ACA的化学成分进行分析,再通过葡聚糖硫酸钠(dextran sodium sulfate,DSS)诱导的小鼠UC模型,评估连续7 d给予柳氮磺吡啶(SASP,200 mg·kg^(-1))和ACA(10、20 mg·kg^(-1))对小鼠体质量、疾病活动指数、结肠长度和结肠病理损伤等指标的影响,并结合体外抗炎活性实验和酶联免疫吸附以及Western blot等实验考察ACA的抗炎活性和机制。结果ACA主要由多种特征型单体生物碱组成。ACA(10、20 mg·kg^(-1))能显著改善DSS诱导的小鼠体质量减轻、疾病指数升高、结肠缩短和病理损伤等。同时,ACA可降低DSS所致的结肠炎症因子(如IL-1β、IL-18和IL-6)的含量。体外炎症模型同样证实ACA具有类似于NOD样受体蛋白3(NLRP3)炎症小体的特异性抑制剂MCC950的抑制LPS和Nigericin双重诱导THP-1细胞上清中上述炎症因子分泌的作用。蛋白表达结果提示该作用可能与其抑制NLRP3信号通路有关。结论ACA含有多种结构类似的单体生物碱,具有抗小鼠UC的活性,可能与抑制NLRP3炎症小体发挥抗炎作用。

OBJECTIVE To investigate the potential therapeutic effects and underlying mechanisms of total alkaloids derived from processed Aconitum carmichaelii Debx(ACA)on ulcerative colitis(UC)in mice.METHODS The chemical composition of ACA was analyzed using liquid chromatography-mass spectrometry.A mouse model of UC was induced using dextran sodium sulfate(DSS)to assess the effects of continuous administration of salicylazosulfapyridine(SASP)(200 mg·kg^(-1))and ACA(10 and 20 mg·kg^(-1))over seven days,evaluating parameters such as body weight,disease activity index,colon length,and pathological damage to the colon.The anti-inflammatory activity and mechanisms of ACA were investigated through in vitro experiments,enzyme-linked immunosorbent assay,and Western blotting.RESULTS ACA is primarily composed of various characteristic monomeric alkaloids.Treatment with ACA(10 and 20 mg/kg)significantly mitigated weight loss,disease index elevation,colon shortening,and pathological damage induced by DSS in the mice.Additionally,ACA reduced the levels of inflammatory factors,including IL-1β,IL-18,and IL-6,elevated by DSS in colitis.The in vitro inflammatory model further demonstrated that ACA functions as a specific inhibitor,MCC950,similar to the NLRP3 inflammasome,which suppresses the secretion of the aforementioned inflammatory factors in the supernatant of LPS-and Nigericin-induced THP-1 cells.Protein expression analysis suggested that this effect may be linked to the inhibition of the NLRP3 signaling pathway.CONCLUSION ACA contains multiple structurally similar monomeric alkaloids that exhibit anti-UC activity in mice,potentially exerting anti-inflammatory effects through the inhibition of the NLRP3 inflammasome.