商陆长期毒性及毒代动力学研究

Study on Long-term Toxicity and Toxicokinetics of Phytolaccae Radix

关键词:  商陆  商陆皂苷甲  肾毒性  “量-时-毒”关系

Key words:  Phytolaccae Radix  esculentoside A  nephrotoxicity  “dose-time-toxicity” relationship

摘要:

目的 通过商陆对大鼠的长期毒性以及毒代动力学的研究，考察商陆“量-时-毒”关系。方法 连续120 d给大鼠灌胃低、中、高剂量商陆水提物，观察大鼠一般状况、体质量、血生化指标、血液学指标、尿液指标及早期损伤指标，观察主要脏器、计算脏器指数并进行病理学检查。并于1，28，120 d各组大鼠静脉取血，用LC-MS/MS测定商陆皂苷甲(esculentoside A，EsA)血药浓度并拟合毒代动力学参数。结果 与空白对照组相比，中、高剂量组大鼠体质量明显降低(P<0.05)、情绪萎靡、活动减少，白细胞含量显著升高(P<0.05或P<0.01)，尿液中尿蛋白显著增高(P<0.05或P<0.01)、肾损伤因子-1含量显著升高(P<0.05或P<0.01)，肾脏指数显著升高(P<0.05)，组织病理学检查直接观察到肾脏出现肾小管损伤与肾间质肾炎，大鼠出现肾脏损伤。毒代动力学研究显示，相同剂量商陆水提物给药时，各组大鼠血浆中EsA的C_max与AUC_0-t随给药时间的延长而升高；相同给药时间点，不同剂量组的大鼠血浆中EsA的C_max与AUC_0-t随剂量的升高而增加。结论 商陆重复给药120 d时具有明显肾毒性，EsA是商陆毒性成分之一；商陆的“量-时-毒”关系为给药剂量越大、给药周期越长，大鼠血浆内EsA的暴露量越高，商陆的肾毒性作用越强。

Abstract:

OBJECTIVE To obtain the “quantity-time-toxicity” relationship by studying the long-term toxic reaction and toxicokinetics of Phytolaccae Radix to rats. METHODS Rats were given low, medium and high doses of aqueous extracts of Phytolaccae Radix for 120 d to observe the general condition, body weight, blood biochemical indexes, hematological indexes, urine indexes, early injury indexes, organ indexes and pathological examinations of the rats. And on 1, 28, and 120 d, blood was collected from each group of rats, and LC-MS/MS method was used to determine the plasma concentration of esculentoside A(EsA) and fit the toxicokinetic parameters. RESULTS Compared with the blank control group, the rats in the middle and high-dose groups were significantly reduced in body weight(P<0.05), sluggish and reduced in activity, white blood cell in the blood were significantly increased(P<0.05 or P<0.01), urine protein in the urine was significantly increased(P<0.05 or P<0.01), kidney injury molecule 1 increased significantly(P<0.05 or P<0.01), kidney-to-body ratio increased significantly(P<0.05), histopathological examination directly observed renal tubular damage and renal interstitial nephritis in the kidneys, and kidney damage in rats. The toxicokinetic study showed that when the same dose was administered, the C_max and AUC_0-t of EsA in the plasma of each group of rats increased with the extension of the administration time. At the same time point of administration, the C_max and AUC_0-t of EsA in the plasma of rats in different doses groups increased with the increase of the dose. CONCLUSION Phytolaccae Radix has obvious nephrotoxicity when it is repeatedly administered for 120 d. EsA is one of the toxic components of Phytolaccae Radix. The relationship of “quantity-time-toxicity” of Phytolaccae Radix is that the greater the dose and the longer the period of administration, the higher the EsA exposure in rat plasma, the stronger the nephrotoxicity of Phytolaccae Radix.