[关键词]
[摘要]
目的研究肾康注射液在正常和慢性肾衰大鼠体内的药动学,为肾康注射液的临床安全合理用药提供科学参考。方法选择黄芪甲苷和大黄酸作为肾康注射液的药动学标记物,建立液相色谱-串联质谱(LC-MS/MS)法检测其血药浓度,并进行专属性、精密度、准确度、回收率、基质效应、稳定性考察;采用连续21 d ig腺嘌呤(250 mg/kg)诱导大鼠慢性肾衰模型,观察并记录两组大鼠的体质量、肾质量、尿量;全自动生化仪检测血清肌酐(Cr)、尿素氮(BUN)水平,观察肾脏外观并进行组织切片HE染色处理。正常和慢性肾衰大鼠静脉滴注肾康注射液(2 mL/kg),采用LC-MS/MS法测定大鼠血浆中黄芪甲苷和大黄酸的血药浓度。结果 LC-MS/MS法采用Hypersil BDS-C18(4.6 mm×150 mm,5 μm)色谱柱;流动相为乙腈-0.1%甲酸水(8:2);体积流量0.8 mL/min;柱温30℃;进样量为10 μL;电喷雾离子源(ESI),负离子扫描检测方式,动态多反应监测模式(MRM)。方法学验证结果表明,LC-MS/MS测定方法快速、灵敏、特异。ig腺嘌呤21 d后,与对照组比较,造模组肾脏肥大,颜色苍白,造模组组织切片的肾小体数量减少、肾小球囊腔增大、肾小管内有棕色的腺嘌呤沉积。与对照组比较,从造模10 d起慢性肾衰大鼠的体质量显著减轻(P<0.05);造模结束后,肾质量显著增加、24 h尿量显著增加、血清肌酐和尿素氮水平显著升高(P<0.05),慢性肾衰大鼠模型成功建立。大鼠药动学实验结果发现,与对照组比较,慢性肾衰大鼠体内黄芪甲苷的药时曲线下面积(AUC0-t)和最大血药浓度(Cmax)分别下降了28.6%和21.6%,黄芪甲苷和大黄酸的清除半衰期(t1/2z)分别缩短了23.5%和43.7%,均具有统计学差异(P<0.05)。结论慢性肾衰大鼠对肾康注射液的清除加快,药物作用时间缩短。
[Key word]
[Abstract]
Objective Pharmacokinetics of astragaloside IV and rhein in normal and chronic renal failure rats were studied after intravenous injection of Shenkang Injection to provide useful information for the clinical rational drug use. Methods Astragaloside IV and rhein were selected as the pharmacokinetic markers of Shenkang Injection. A liquid chromatography tandem mass spectrometry (LC-MS/MS) method was established to detect the plasma concentration of Shenkang injection, and the specificity, precision, accuracy, recovery, matrix effect and stability were investigated. Chronic renal failure was induced by adenine (250 mg/kg) in rats. Rats received Shenkang Injection (2 mL/kg) intravenously. The body weight, kidney weight and urine volume of the two groups were observed and recorded, and the levels of serum creatinine (Cr) and urea nitrogen (BUN) were detected by automatic biochemical analyzer, and the appearance of kidney was observed and the tissue sections were stained with HE. Plasma concentrations of astragaloside IV and rhein were determined by LC-MS/MS. Results Hypersil BDS-C18 (4.6 mm×150 mm, 5 μm) column was used. The mobile phase was acetonitrile-0.1% formic acid water (8: 2); volume flow rate was 0.8 mL/min; column temperature 30 ℃; injection volume was 10 μL; electrospray ion source (ESI), negative ion scanning mode, dynamic multi reaction monitoring mode (MRM). The results showed that LC-MS/MS was rapid, sensitive and specific. After ig adenine for 21 days, compared with control group, the kidney of the model group was hypertrophy and pale, the number of renal corpuscles in the tissue section of model group was reduced, the glomerular capsule cavity was enlarged, and there was brown adenine deposition in the renal tubules. Compared with control group, the body weight of CRF rats was significantly reduced from the 10th day of modeling (P < 0.05). After modeling, the kidney weight, 24 h urine volume, serum creatinine and urea nitrogen levels were significantly increased (P < 0.05), and the CRF rat model was successfully established. In CRF rats, area under the plasma concentration-time curve (AUC0-t) and maximum plasma concentration (Cmax) of astragaloside IV were significantly decreased by 28.6% and 21.6%, respectively (P < 0.05). The half-life (t1/2z) of astragaloside IV and rhein were significantly reduced by 23.5% and 43.7%, respectively (P < 0.05). Conclusion Shenkang Injection exhibited an accelerated clearance in CRF rats, which might shorten effective period.
[中图分类号]
R969.1
[基金项目]
国家自然科学基金资助项目(81302838,81903706)