Background: Hypecoum leptocarpum Hook. f. et Thoms., which is used in traditional Tibetan medicine as an antipyretic, antitussive, analgesic, and anti-inflammatory agent, contains a variety of alkaloids that could be responsible for its analgesic and anti-inflammatory properties. Objective: The present study was designed to investigate the anti-inflammatory activity of the total alkaloids from H. leptocarpum (AHL) in vitro and to elucidate the chemical structure of the anti-inflammatory components in AHL. Materials and Methods: Chemical characterization was performed using liquid chromatography/quadrupole-time-of-flight mass and diode-array detector-high performance liquid chromatography. The anti-inflammatory effects of AHL were investigated by measuring the production of inflammatory cytokines using enzyme-linked immunosorbent assay and mRNA expression by real-time polymerase chain reaction in lipopolysaccharide-induced RAW 264.7 macrophages. Results: Chemical analysis of AHL revealed the presence of seven alkaloids, protopine (13.3%), cryptopine (1.5%), leptopidinine, leptocarpine, corydamine, dihydroleptopine, and oxohydrastinine. AHL significantly suppressed the production of nitric oxide (NO), interleukin-1 beta (IL-1 β), IL-6, and tumor necrosis factor-alpha (TNF-α) in LPS-induced RAW 264.7 cells. The maximum levels of suppression of NO, IL-1 β, IL-6, and TNF-α were 86.8% ± 2.2%, 70.1% ± 1.5%, 100.1% ± 2.5%, and 50.8% ± 3.6%, respectively. IC50values of suppression of cytokine production by AHL were 7.47 ± 2.81 μg/mL (NO), 0.12 ± 0.28 μg/mL (IL-1 β), 0.56 ± 0.37 μg/mL (IL-6), and 18.95 ± 5.23 μg/mL (TNF-α). AHL was also shown to downregulate mRNA expression of inducible NO synthase, IL-1 β, IL-6, and TNF-α in vitro. Conclusion: The study provides convincing evidence that AHL has strong anti-inflammatory activity. The potent activity is likely a result of synergy between the different alkaloids. Abbreviations used: The total alkaloids from H. leptocarpum: AHL; Nitric oxide: NO; Interleukin-1 beta IL-1β; Interleukin-6: IL-6; Tumor necrosis factor-alpha: TNF-α; Prostaglandin E2: PGE2; Inducible nitric oxide synthase: iNOS; Nonsteroidal anti-inflammatory drugs: NSAIDs; lipopolysaccharide: LPS; The total ion chromatograms: TIC; The liquid chromatography/quadrupole-time of flight: LC/Q-TOF; Nuclear factor-kappa B: NF-κB; Janus kinase-signal transducers and activators of transcription: JAK-STAT.

中文翻译：

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Hypecoum leptocarpum 钩总生物碱的抗炎活性。F。汤姆斯

背景：Hypecoum leptocarpum Hook。F。et Thoms. 在传统藏药中用作解热、镇咳、镇痛和抗炎剂，含有多种生物碱，可能是其镇痛和抗炎特性的原因。目的：本研究旨在研究细果总生物碱（AHL）的体外抗炎活性，并阐明AHL中抗炎成分的化学结构。材料和方法：使用液相色谱/四极杆飞行时间质量和二极管阵列检测器-高效液相色谱进行化学表征。通过使用酶联免疫吸附试验测量炎性细胞因子的产生和通过脂多糖诱导的 RAW 264.7 巨噬细胞中的实时聚合酶链反应测量 mRNA 表达来研究 AHL 的抗炎作用。结果：AHL 的化学分析显示存在 7 种生物碱，即原平 (13.3%)、隐品碱 (1.5%)、瘦多碱、细果芸香碱、苯丁胺、二氢瘦素和氧代尿嘧啶。AHL 显着抑制 LPS 诱导的 RAW 264.7 细胞中一氧化氮 (NO)、白细胞介素 1 β (IL-1 β)、IL-6 和肿瘤坏死因子-α (TNF-α) 的产生。NO、IL-1 β、IL-6 和 TNF-α 的最大抑制水平分别为 86.8% ± 2.2%、70.1% ± 1.5%、100.1% ± 2.5% 和 50.8% ± 3.6%。AHL 抑制细胞因子产生的 IC50 值为 7.47 ± 2.81 μg/mL (NO)，0.12 ± 0.28 μg/mL (IL-1 β)、0.56 ± 0.37 μg/mL (IL-6) 和 18.95 ± 5.23 μg/mL (TNF-α)。AHL 还显示在体外下调诱导型 NO 合酶、IL-1 β、IL-6 和 TNF-α 的 mRNA 表达。结论：该研究提供了令人信服的证据，表明 AHL 具有很强的抗炎活性。有效的活性可能是不同生物碱之间协同作用的结果。使用的缩写：H. leptocarpum 的总生物碱：AHL；一氧化氮：NO；白细胞介素-1 β IL-1β；白细胞介素6：IL-6；肿瘤坏死因子-α：TNF-α；前列腺素 E2：PGE2；诱导型一氧化氮合酶：iNOS；非甾体抗炎药：非甾体抗炎药；脂多糖：LPS；总离子色谱图：TIC；液相色谱/四极杆-飞行时间：LC/Q-TOF；核因子-κB：NF-κB；