天然产物研究与开发 ›› 2024, Vol. 36 ›› Issue (5): 825-837.doi: 10.16333/j.1001-6880.2024.5.012

• 开发研究 • 上一篇    下一篇

基于炎症大鼠代谢组学的马钱子炮制减毒存效机制研究

秦伟瀚,魏江平,张小梅,阳   勇*   

  1. 重庆市中药研究院,重庆 400065
  • 出版日期:2024-05-28 发布日期:2024-05-29
  • 基金资助:
    国家重大新药创制项目(2017ZX09101002-002-004);重庆市卫生健康委项目(2021ZY023651)

Study on mechanism of attenuating toxicity and preserving effect of Strychnine Semen processing based on metabolomics in inflammatory rats

QIN Wei-han,WEI Jiang-ping,ZHANG Xiao-mei,YANG Yong*   

  1. QIN Wei-han,WEI Jiang-ping,ZHANG Xiao-mei,YANG Yong*
  • Online:2024-05-28 Published:2024-05-29

摘要:

运用动物模型、血浆药物化学及代谢组学方法,探讨马钱子炮制前后干预炎症大鼠作用机制。按不同剂量灌胃小鼠,以死亡率初步考察炮制前后毒性;建立大鼠蛋清性足跖肿胀模型,通过肿胀率和炎性因子水平评价炮制前后抗炎效果;采用高分辨质谱结合多元统计学,鉴定入血后的移行成分及相应的代谢产物;比较分析各实验组代谢轮廓及差异生物标志物,并构建其代谢通路。结果显示,高剂量下马钱子砂烫品的毒性要明显低于生品。生品和砂烫品均能显著降低大鼠足肿胀率和肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)、白介素-6(IL-6)的水平,炮制前后在抗炎效果上差异无统计学意义(P>0.05)。通过血浆药物分析,鉴定出了12个移行成分,以及相应60个代谢产物,S-plot鉴定出10个差异性化合物。代谢组学实验共发现10个内源性代谢物含量发生显著变化(P<0.05),生物标志物主要涉及苯丙氨酸代谢、甘油磷脂代谢、谷氨酸及谷氨酰胺循环、精氨酸和脯氨酸代谢等途径。通过本研究确证马钱子生品和砂烫品均有较好抗炎治疗作用,高温炮制可以降低马钱子毒性;入血后有效成分应多为生物碱,经过I相反应就容易被体内清除;马钱子炮制抗炎、镇痛、减毒机制主要与氨基酸和脂质代谢相关。该实验结果进一步阐明了起效物质基础及体内作用机制,为后续炮制工艺改进和新药研发等提供科学支撑。

关键词: 马钱子, 炮制, 抗炎作用, 代谢组学, 减毒存效

Abstract:

Using the methods of animal model,plasma pharmacochemistry and metabolomics,the mechanism of intervention of Strychnine Semen before and after processing inflammation in rats was investigated.The toxicity before and after processing was preliminarily investigated according to the mortality of mice given different doses.The rat model of egg white foot swelling was established,and the anti-inflammatory effect was evaluated by swelling rate and inflammatory factor level before and after processing.High resolution mass spectrometry combined with multivariate statistics were used to identify the migratory components and their metabolites.The metabolic profiles and different biomarkers of each experimental group were compared and analyzed,and their metabolic pathways were constructed.The results showed that the toxicity of sand-scorch product at high dose was significantly lower than that of raw product.Both raw and sand-scorch products could significantly reduce the swelling rate and the levels of tumor necrosis factor-α (TNF-α),interleukin-1β (IL-1β) and interleukin-6 (IL-6) of rats′ feet,and there was no statistical difference in anti-inflammatory effect before and after processing (P>0.05).Through plasma drug analysis,12 transitional components and corresponding 60 metabolites were identified,and 10 different compounds were identified by S-plot.Metabolomics experiments found significant changes in the content of ten endogenous metabolites (P<0.05).The biomarkers mainly involved phenylalanine metabolism,glycerophospholipid metabolism,glutamic acid and glutamine cycle,arginine and proline metabolism and other pathways.This study confirmed that both raw and sand-scorch products of Strychnine Semen had good anti-inflammatory effects,and high temperature processing could reduce the toxicity of Strychnine Semen.After entering the blood,the active ingredients of the sand-scorch products should mostly be alkaloids,and they are easily eliminated by the body after I reaction.The difference analysis can indirectly prove that the anti-inflammatory effect of the sand-scorch products is higher than that of the raw products.The anti-inflammatory,analgesic and attenuated mechanisms of sand-scorch products are mainly related to amino acid metabolism and lipid metabolism.The experimental results further elucidated the active substance basis and the mechanism of action in vivo,and provided scientific support for the subsequent improvement of processing technology and the development of new drugs.

Key words: Strychnine Semen, processing, anti-inflammatoryeffect, metabolomics, attenuating toxicity and preserving effect

中图分类号:  R917