Sun Dongdong, Yan Qiuying, Xu Xiaofang, Shen Weixing, Xu Changliang, Tan Jiani, Zhang Haibin, Li Liu, Cheng Haibo
a Translational Medicine Center of Nanjing University of Chinese Medicine , Nanjing , China.
b Jiangsu Collaborative Innovation Center of Traditional Chinese Medicine (TCM) Prevention and Treatment of Tumor , Nanjing , China.
Pharm Biol. 2017 Dec;55(1):937-945. doi: 10.1080/13880209.2017.1285327.
BushenHuoxue decoction (BSHXD) is a Chinese medicine prescription, which is composed of nine Chinese medical materials, used to treat osteoarthritis (OA).
This study develops sensitive and convenient LC-MS/MS methods to analyze chemical components from BSHXD, and assess the anti-inflammatory activities thereof.
The chemical composition from BSHXD water extract was qualitative analyzed by high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (HPLC-ESI-Q-TOF-MS). Twelve reference compounds were analyzed by UPLC-ESI-MS/MS. Anti-inflammatory activities of target components were assessed by ELISA at 20 and 100 μg/mL.
It is the first time that 88 compounds were qualitatively identified from BSHXD, of which 12 with potential in treating OA according to the literature were quantified. Within BSHXD the contents of quercetin, isopsoralen, icarisideII, osthole, and isoimperatorin increased remarkably compared with those in single herb which make up BSHXD, the contents were 0.1999, 0.4634, 0.0928, 0.5364, and 0.1487 mg/g. ELISA data displayed that BSHXD and the five compounds mentioned inhibited the expressions of TNF-α, IL-6 and NO released from LPS-stimulated RAW264.7 cell, with maximum inhibition rates of 104.05% (osthole, 100 μg/mL), 100.03% (osthole, 100 μg/mL), and 93.46% (isopsoralen, 20 μg/mL), respectively.
Content changes of 12 compounds in BSHXD and single herbs which comprise the prescription were measured and analyzed. Contents of five compounds increased may be explained by solubilization between drugs and chemical reaction. ELISA results reported that the increased contents of the five compounds could inhibit expression of the inflammatory factors.
补肾活血汤（BSHXD）是一种中药方剂，由九味中药材组成，用于治疗骨关节炎（OA）。
本研究建立灵敏便捷的液相色谱-串联质谱（LC-MS/MS）方法来分析补肾活血汤中的化学成分，并评估其抗炎活性。
采用高效液相色谱-电喷雾电离四极杆飞行时间质谱（HPLC-ESI-Q-TOF-MS）对补肾活血汤水提取物的化学成分进行定性分析。通过超高效液相色谱-电喷雾串联质谱（UPLC-ESI-MS/MS）分析12种对照品。采用酶联免疫吸附测定（ELISA）法在20和100μg/mL浓度下评估目标成分的抗炎活性。
首次从补肾活血汤中定性鉴定出88种化合物，其中根据文献有12种具有治疗骨关节炎的潜力并进行了定量分析。与组成补肾活血汤的单味药材相比，补肾活血汤中槲皮素、异补骨脂素、淫羊藿苷Ⅱ、蛇床子素和异欧前胡素的含量显著增加，含量分别为0.1999、0.4634、0.0928、0.5364和0.1487mg/g。ELISA数据显示，补肾活血汤及上述5种化合物抑制脂多糖（LPS）刺激的RAW264.7细胞释放肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）和一氧化氮（NO）的表达，最大抑制率分别为104.05%（蛇床子素，100μg/mL）、100.03%（蛇床子素，100μg/mL）和93.46%（异补骨脂素，20μg/mL）。
测定并分析了补肾活血汤及组成该方剂的单味药材中12种化合物的含量变化。5种化合物含量增加可能是由于药物间的增溶作用和化学反应所致。ELISA结果表明，这5种化合物含量增加可抑制炎症因子的表达。
