Department of Pharmacy, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
School of Minerals Processing and Bioengineering, Central South University, Changsha, China.
Biopharm Drug Dispos. 2023 Dec;44(6):406-419. doi: 10.1002/bdd.2377. Epub 2023 Sep 7.
The study aimed to explore the pharmacokinetic and pharmacodynamic alterations of the active components of Shenkang injection (i.e. hydroxy saffron yellow pigment A [HSYA], tanshinol, rheum emodin, and astragaloside IV) in rats with chronic renal failure (CRF), and establish a pharmacokinetic-pharmacodynamic model (PK-PD model) in order to provide a scientific and theoretical basis for the rational clinical use of Shenkang injection. Sprague-Dawley (SD) rats were randomly divided into a normal group, model group, and Shenkang injection group. A rat model of CRF was induced by adenine gavage and then followed by drug administration via tail vein injection. Orbital blood was collected at different timepoints and the blood concentrations of the four active components were measured by UHPLC-Q-Orbitrap HRMS. Serum levels of creatinine (Scr), urea nitrogen (BUN), and uric acid (UA) were determined using an automatic biochemical analyzer. A PK-PD model was established, and DAS 3.2.6 software was used for model fitting as well as statistical analysis. TGF-β1 was utilized to induce normal rat kidney cells to construct a renal fibrosis model to investigate the protective effect of the pharmacological components on renal fibrosis. The pharmacokinetic analysis of hydroxy saffron yellow pigment A, tanshinol, rheum emodin, and astragaloside IV based on UHPLC-Q-Orbitrap HRMS was stable. The linear regression equations for the four active components were as follows: Y = 0.031X + 0.0091 (R = 0.9986) for hydroxy saffron yellow pigment A, Y = 0.0389X + 0.164 (R = 0.9979) for tanshinol, Y = 0.0257X + 0.0146 (R = 0.9973) for rheum emodin, and Y = 0.0763X + 0.0139 (R = 0.9993) for astragaloside IV, which indicated good linear relationships. The methodological investigation was stable, with the interday and intraday precision RSD <10%. Meanwhile, the recoveries ranged between 90% and 120%, in accordance with the requirements for in vivo analysis of drugs. Compared with the model group, the levels of Scr, BUN, and UA were significantly decreased after 20 min in the Shenkang injection group (p < 0.01). The PK-PD model showed that the four active components in the Shenkang injection group could fit well with the three effect measures (i.e. Scr, BUN, and UA), with the measured values similar to the predicted values. The cell model of renal fibrosis showed that the connective tissue growth factor and FN1 protein expression levels were significantly lower in the Shenkang injection group than those in the model group, and the cell fibrosis was improved. The established method for in vivo analysis of Shenkang injection was highly specific, with good separation of the components and simple operation. The total statistical moment could well integrate the pharmacokinetic parameters of the four active components. After treatment with Shenkang injection, all indexes in the administered group improved and showed significant inhibition of renal cell fibrosis in vitro. This study could provide scientific reference ideas for the clinical rational use of traditional Chinese medicine.
本研究旨在探讨肾康注射液（即羟基红花黄色素 A[HSYA]、丹参醇、大黄素和黄芪甲苷 IV）在慢性肾衰竭（CRF）大鼠体内的药代动力学和药效学变化，并建立药代动力学-药效学模型（PK-PD 模型），为肾康注射液的临床合理应用提供科学理论依据。SD 大鼠随机分为正常组、模型组和肾康注射液组。通过灌胃腺嘌呤诱导大鼠 CRF 模型，然后通过尾静脉注射给药。在不同时间点采集眶内血，采用 UHPLC-Q-Orbitrap HRMS 测定 4 种活性成分的血药浓度。采用自动生化分析仪测定血清肌酐（Scr）、尿素氮（BUN）和尿酸（UA）水平。采用 DAS 3.2.6 软件建立 PK-PD 模型并进行模型拟合和统计分析。利用 TGF-β1 诱导正常大鼠肾细胞构建肾纤维化模型，探讨药理成分对肾纤维化的保护作用。基于 UHPLC-Q-Orbitrap HRMS 的羟基红花黄色素 A、丹参醇、大黄素和黄芪甲苷 IV 的药代动力学分析稳定。四种活性成分的线性回归方程如下：羟基红花黄色素 A 的 Y = 0.031X + 0.0091（R² = 0.9986），丹参醇的 Y = 0.0389X + 0.164（R² = 0.9979），大黄素的 Y = 0.0257X + 0.0146（R² = 0.9973），黄芪甲苷 IV 的 Y = 0.0763X + 0.0139（R² = 0.9993），表明具有良好的线性关系。方法学考察稳定，日内和日间精密度 RSD<10%。同时，回收率在 90%~120%之间，符合药物体内分析的要求。与模型组相比，肾康注射液组 Scr、BUN 和 UA 水平在 20 min 时明显降低（p<0.01）。PK-PD 模型显示，肾康注射液组的四种活性成分均能很好地拟合三个效应指标（即 Scr、BUN 和 UA），实测值与预测值相似。肾纤维化细胞模型显示，肾康注射液组结缔组织生长因子和 FN1 蛋白表达水平明显低于模型组，细胞纤维化得到改善。建立的肾康注射液体内分析方法具有高度特异性，各成分分离良好，操作简单。总统计矩可很好地整合四种活性成分的药代动力学参数。经肾康注射液治疗后，给药组各项指标均有改善，体外对肾细胞纤维化有明显抑制作用。本研究可为中药临床合理应用提供科学参考思路。
