基于大鼠室性早搏心脏毒性的附子质量生物评价方法研究

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附子是确有疗效的毒剧类中药,构建关联临床毒副反应的质量评价方法对提高其炮制减毒质控水平和保障临床安全性具有重要意义。该文针对附子可导致室性早搏这一早期心脏中毒反应,构建基于大鼠室性早搏(premature ventricular contractions,PVC)最小中毒量(minimal toxic dose,MTD)测定的附子炮制品质量生物评价方法,通过影响因素考察优化方法学(动物性别、体重、对照品稳定性与供试品稳定性),检测结果的重现性符合药品质量生物评价要求。应用该方法检测生附子及其不同炮制品黑顺片、白附片、蒸附片、刨附片、炮天雄致大鼠室性早搏MTD,结果表明,生附子炮制后MTD均显著升高(P<0.05),其中黑顺片、蒸附片、刨附片与白附片的MTD分别为生附子的15.76,22.36,19.65,20.97倍;而炮天雄几乎无毒,用该方法检测不出MTD。该方法可以比较好的反映附子及其炮制品的整体心脏毒性。进一步结合附子炮制品的化学成分分析,发现炮制品中双酯型生物碱乌头碱、新乌头碱、次乌头碱,以及单酯型生物碱苯甲酰乌头原碱与PVC呈显著相关。此外,可能还存在其他与致心律失常相关的未测成分,值得深入研究。上述研究为附子临床早期心脏毒性评价与质量控制提供了整体、综合的方法,可作为化学评价方法的有益补充,为保障附子临床应用的安全性提供了新的方法。 Aconite is really a curative effect of traditional Chinese medicine, to build the quality of associated clinical adverse reactions to evaluate the quality of its processing of attenuated quality control and protection of clinical safety is of great significance. This article aimed at the early cardiotoxicity induced by aconite, which could lead to premature ventricular contractions, and established a biological evaluation method of aconite product quality based on the determination of minimal toxic dose (MTD) of premature ventricular contractions (PVC) The optimization methodology (animal sex, body weight, reference stability and test sample stability) was investigated by influencing factors, and the reproducibility of the test results was in line with the requirements of biological quality evaluation of drugs. The method was used to detect the MTD of ventricular premature beat in rats treated with Radix Aconiti Preparata and its different processed products. The MTD of ventricular premature beats in rats was significantly increased (P <0.05). The MTDs of black tablets, steamed tablets, plaque tablets and white tablets were 15.76, 22.36, 19.65 and 20.97 times higher than that of raw materials, while Gutianxiong was almost nontoxic and was detected by this method No MTD. This method can better reflect the overall cardiotoxicity of aconite and its processed products. Further analysis of the chemical composition of the aconite processed products revealed that the doubly-esterified alkaloids aconitine, mesaconitine, hypaconitine, and monoester alkaloid benzoyl aconitine in the processed product were significantly different from PVC Related. In addition, there may be other unmeasured components related to arrhythmia, it is worth further study. The above studies provide an overall and comprehensive method for the early clinical evaluation and quality control of aconite, which can be used as a useful complement to the chemical evaluation methods and provide a new method for the safety of aconite in clinical application.
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