目的 探究含羞草Mimosa pudica根水提物对干酵母致发热模型大鼠的解热作用及其药效物质基础。方法 采用HPLC法指认含羞草根水提物的化学成分。通过背部sc 20%干酵母混悬液建立大鼠发热模型，ELISA法测定大鼠血清中肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素-1β（interleukin-1β，IL-1β）、IL-6及下丘脑中5-羟色胺（5-hydroxytryptamine，5-HT）、前列腺素E₂（prostaglandin E₂，PGE₂）、环磷酸腺苷（cyclic adenosine monophosphate，cAMP）水平，采用超分辨率液质联用技术对大鼠血清和下丘脑进行非靶向代谢组学分析，以阐明含羞草根水提物的解热作用机制。采用柱色谱和半制备液相并结合NMR、MS等现代波谱技术对含羞草根化学成分进行分离与鉴定，采用分子对接评价含羞草根化学成分与解热关键靶点Toll样受体4（Toll-like receptor 4，TLR4）的亲和力，并通过考察含羞草根化学成分对脂多糖诱导RAW264.7细胞一氧化氮（nitric oxide，NO）释放量的影响评价其抗炎活性。结果 4种黄酮类成分（槲皮素、芦丁、杨梅素、木犀草素）在含羞草根水提物中被指认。药效学实验表明在造模5 h后大鼠体温明显升高（P＜0.01），含羞草根水提物组大鼠体温在各个时段均显著低于模型组（P＜0.01），含羞草根显著降低发热大鼠下丘脑中cAMP、5-HT、PGE₂和血清中TNF-α、IL-6、IL-1β水平（P＜0.001）。代谢组学结果显示干酵母致大鼠发热后下丘脑及血清的代谢轮廓发生了显著的变化，分别鉴定出34、16个潜在的生物标志物；经含羞草根干预后，以上生物标志物含量显著回调，主要影响氨基酸代谢、嘌呤代谢、嘧啶代谢、柠檬酸循环、维生素B₆代谢、初级胆汁酸生物合成、类固醇激素生物合成等途径。从含羞草根中分离出8个有机酸类成分，分别鉴定为二十六烷（1）、十七烷酸（2）、(2S)-1-O-三十七烷酰基甘油（3）、二氢咖啡酸甲酯（4）、3,4-二羟基苯酰甲醇（5）、原儿茶酸（6）、咖啡酸（7）、没食子酸乙酯（8）。其中，除化合物7外，其余化合物均为含羞草属植物首次发现。分子对接结果显示除化合物1和3外，其余化合物和黄酮类成分与TLR4有一定的亲和性。化合物1～8均具有较强的体外抗炎作用，可能为含羞草根发挥解热作用的物质基础。结论 含羞草水提物对干酵母致热大鼠有显著的解热作用，其作用机制可能与抑制致热内源性因子的释放和调节氨基酸代谢、嘌呤代谢、能量代谢等途径有关，其发挥解热的药效物质基础可能为有机酸类成分和黄酮类成分。

Objective To investigate the antipyretic effects and pharmacological substance basis of water extract of root of Mimosa pudica on a fever model rat induced by dry yeast. Methods The chemical constituents of water extract of root of M. pudica were identified using HPLC. A rat fever model was established by subcutaneous injection of 20% dry yeast suspension. The levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and IL-6 in serum, as well as 5-hydroxytryptamine (5-HT), prostaglandin E₂ (PGE₂) and cyclic adenosine monophosphate (cAMP) in hypothalamus were measured using ELISA. Non-targeted metabolomics analysis of serum and hypothalamus in rats was performed using UPLC-Orbitrap Fusion MS to elucidate the antipyretic mechanism of water extract of root of M. pudica. Column chromatography and semi-preparative liquid chromatography, combined with modern spectroscopic techniques such as NMR and MS, were employed to isolate and identify the chemical constituents of root of M. pudica. Molecular docking was used to evaluate the affinity of the identified chemical constituents for the key antipyretic target Toll-like receptor 4 (TLR4). And its anti-inflammatory activity was evaluated by examining the effect of chemical components in root of M. pudica on lipopolysaccharide induced nitric oxide (NO) release in RAW264.7 cells. Results A total of four flavonoid components (quercetin, rutin, myricetin, luteolin) were identified in water extract of root of M. pudica. Pharmacological experiments showed that the body temperature of rats significantly increased 5 h after modeling (P < 0.01), water extract of root of M. pudica group exhibited significantly lower body temperatures at all time points compared to model group (P < 0.01), the root of M. pudica significantly reduced the levels of cAMP, 5-HT, PGE₂ in hypothalamus and TNF-α, IL-6, IL-1β in serum of feverish rats (P < 0.001). Metabolomics results revealed significant changes in metabolic profiles of hypothalamus and serum after dry yeast-induced fever, identifying 34 and 16 potential biomarkers, respectively. After intervention with root of M. pudica, the levels of these biomarkers were significantly regulated, primarily affecting pathways such as amino acid metabolism, purine metabolism, pyrimidine metabolism, citric acid cycle, vitamin B₆ metabolism, primary bile acid biosynthesis and steroid hormone biosynthesis. A total of eight organic acid components were isolated from root of M. pudica and identified as hexacosane (1), heptadecanoic acid (2), (2S)-1-O-heptatriacontanoylglycerol (3), methyl dihydrocaffeate (4), 3,4-dihydroxybenzyl alcohol (5), protocatechuic acid (6), caffeic acid (7) and ethyl gallate (8). Among these, compounds 1—8, except for compound 7, were discovered for the first time in Mimosa genus. Molecular docking results showed that except for compounds 1 and 3, the remaining compounds and flavonoids exhibited affinity for TLR4. Compounds 1—8 demonstrated strong anti-inflammatory effects in vitro, suggesting their potential as the material basis for the antipyretic effects of root of M. pudica.Conclusion The water extract of root of M. pudica exhibits significant antipyretic effects on dry yeast-induced fever in rats. Its mechanism may be related to the inhibition of endogenous pyrogenic factor release and the regulation of metabolic pathways such as amino acid metabolism, purine metabolism, and energy metabolism. The pharmacological substance basis responsible for its antipyretic effects are likely organic acids and flavonoids.

R285.5

国家中医药管理局中医药传承与创新“百千万”人才工程-岐黄工程首席科学家支持项目（国中医药人教函[2021]7号）；2022年全国名老中医药专家传承工作室建设项目（国中医药人教函[2022]75号）；第七批全国老中医药专家学术经验继承工作项目（国中医药人教函[2022]76号）；黑龙江省“头雁”团队支持项目（黑龙江省头雁行动领导小组文件[2019]5号）；全国老药工传承工作室建设项目（国中医药人255号）；广东省基础与应用基础研究基教函[2024]金（2023A1515110740）