柴胡解表退热功效相关生物活性指标与柴胡皂苷a、柴胡皂苷d含量相关性分析

目的：寻找与柴胡解表退热功效相关的生物活性指标，并探明此功效的生物活性量化值与柴胡皂苷a(SSa)和柴胡皂苷d (SSd)含量是否具有相关性。方法：采用临床煎药方式获得柴胡水煎液，并通过醇沉去除多糖和蛋白质等大分子物质，得到适宜体外实验的柴胡提取物。从抗炎、抗过敏、抗氧化应激及解热这4类与柴胡解表退热功效相关的体外模型中筛选量效关系明确的生物活性指标，并赋予各项指标不同的权重系数，对7个柴胡样品在各项指标上的作用强弱进行排序打分；分数与相应指标权重的积为该项得分；每项得分之和为该样品解表退热之生物活性量化值；采用CCK-8法检测细胞活力；Griss法检测细胞上清液中一氧化氮（NO）水平；ELISA法测定细胞上清液中肿瘤坏死因子-α (TNF-α)及补体活化水平；铁离子还原能力法（FRAP）测定总抗氧化能力；高效液相色谱法测定柴胡中的SSa和SSd含量。结果：筛选得到4项关联柴胡解表退热功效的量效关系明确的生物活性指标，分别为脂多糖（LPS）诱导RAW264.7细胞分泌NO、LPS诱导THP-1细胞分泌TNF-α、总抗氧化能力及酵母多糖介导的替代途径补体活化。相关性分析显示，7个样品的S...

Bioactive Indicators Related to Actions of Relieving the Exterior and Reducing the Fever of Bupleuri Radix and Their Correlation with Content of Saikosaponin a and Saikosaponin d

Objective To explore the bioactive indicators related to the actions of relieving the exterior and reducing the fever of Bupleuri Radix (BR) and study the correlation between the bioactive quantized value and saikosaponin a (SSa) and saikosaponin d (SSd) content.Methods The BR decoction was obtained clinically, and the macromolecular substances such as polysaccharide and protein were removed by alcohol precipitation, so as to obtain BR extract suitable for in vitro experiments. This study selected bioactive indicators with definite concentration-effect relationships from four types of in vitro models, including anti-inflammatory, anti-allergic, anti-oxidative stress, and antipyretic effects, which are associated with the actions of relieving the exterior and reducing the fever of BR. In addition, different weight coefficients were assigned, and the strength of the effects of seven samples of BR was scored based on the selected indicators. The score multiplied by the corresponding weight coefficient equaled the value of each indicator, and the sum of four values was regarded as a bioactive quantized value for each sample in relieving the exterior and reducing the fever. Cell viability was determined by CCK-8 assay. Nitric oxide (NO) levels in the cell supernatant were measured using the Griess method. The levels of TNF-α and in vitro complement activation assay in the cell supernatant were determined by enzyme-linked immunosorbent assay (ELISA). Total antioxidant capacity was assessed using the ferric ion reducing antioxidant power (FRAP) assay. SSa and SSd contents in BR were analyzed using high-performance liquid chromatography (HPLC).Results Four bioactive indicators related to the actions of relieving the exterior and reducing the fever of BR were selected: NO production of lipopolysaccharide (LPS)-activated RAW264.7 cells, total antioxidant capacity, TNF-α production of LPS-activated THP-1 cells, and complement activation induced by yeast polysaccharide. As a result, there was no correlation between the bioactive quantized values and the total content of SSa and SSd of the seven samples (P>0.05).Conclusion The actions of relieving the exterior and reducing the fever of BR are related to the inhibition of proinflammatory factor (NO) production, antioxidant properties, production of endogenous pyrogen (TNF-α), and complement C5a activation. In addition, the quality control substances (SSa and SSd) of BR in the Chinese Pharmacopoeia (2020) may not represent this efficacy.