目的 建立基于抗炎生物效应的淫羊藿质量评价方法。方法 采用免疫刺激性DNA（immune-stimulatory DNA，ISD）诱导的小鼠骨髓来源巨噬细胞（bone marrow-derived macrophages，BMDMs）为炎症细胞模型，通过检测磷酸化干扰素基因刺激因子（phosphorylated stimulator of interferon genes，p-STING）、磷酸化干扰素调节因子3（phosphorylated interferon regulatory factor 3，p-IRF3）蛋白表达以及I型干扰素（type I interferon，IFN-β）表达并结合细胞活力测定，初步验证淫羊藿的抗炎作用。以STING抑制剂H151为阳性对照药，建立以IFN-β抑制率为评价指标的抗炎生物效价测定方法，评价不同品种和批次淫羊藿样品的抗炎效价。采用高效液相色谱仪测定5个品种15批淫羊藿样品中6个特征成分（双藿苷A、朝藿定A、朝藿定B、朝藿定C、淫羊藿苷、宝藿苷Ⅰ）的含量，再通过相关性分析方法探讨抗炎效应与特征成分含量之间的关联性。结果 淫羊藿提取物可以抑制环磷酸鸟苷-腺苷酸合成酶（cyclic GMP-AMP synthase，cGAS）-STING信号通路活化指标p-STING、p-IRF3以及IFN-β的表达，证明其具有良好的抗炎效应。15批淫羊藿的抗炎效价为221.05～1130.50 U/mL，不同品种的平均抗炎效价从高到低排序为箭叶淫羊藿（951.17 U/mL）、朝鲜淫羊藿（748.38 U/mL）、淫羊藿（746.71 U/mL）、柔毛淫羊藿（728.18 U/mL）、粗毛淫羊藿（528.87 U/mL），表明箭叶淫羊藿具有良好的抗炎生物效应，且伪品粗毛淫羊藿比4个正品抗炎效价都低。相关性分析显示，淫羊藿苷与淫羊藿抗炎效价呈显著正相关（r＝0.540，P＜0.05），说明该成分可能在抗炎活性中起主要作用。结论 建立了基于抗炎生物效价的淫羊藿质量评价方法，其能够直接反映不同淫羊藿在抗炎方面的质量“优劣”，为具有抗炎性质的中药质量评价提供参考。

Objective To establish a method for evaluating the quality of Yinyanghuo (Epimedii Folium) based on anti-inflammatory biological potency. Methods Immune-stimulatory DNA (ISD)-induced mouse bone marrow-derived macrophages (BMDMs) were used as an inflammatory cell model, and the anti-inflammatory effects of Epimedii Folium were preliminarily verified by detecting the protein expression levels of phosphorylated stimulator of interferon genes (p-STING), phosphorylated interferon regulatory factor 3 (p-IRF3) and the expression of type I interferon (IFN-β) combined with cell viability assay. Using STING inhibitor H151 as a positive control, an anti-inflammatory biological potency assay was established with IFN-β inhibition rate as the evaluation index to evaluate the anti-inflammatory potency of Epimedii Folium samples of different varieties and batches. High performance liquid chromatography was used to determine the contents of six characteristic components (diphylloside A, epimedin A, epimedin B, epimedin C, icariin, baohuoside I) in 15 batches of Epimedii Folium samples of five varieties, and then exploring the correlations between anti-inflammatory potency and the contents of the characteristic components by correlation analysis methods. Results The extract of Epimedii Folium could inhibit the expression levels of p-STING, p-IRF3 and IFN-β, which were the activation indexes of cyclic GMP-AMP synthase (cGAS)-STING signaling pathway, proving that it had a good anti-inflammatory effect. The anti-inflammatory potency of 15 batches of Epimedii Folium samples ranged from 221.05 to 1 130.50 U/mL , and the average anti-inflammatory potency of different varieties in order of from high to low was Epimedium sagittatum (951.17 U/mL), E. koreanum (748.38 U/mL), E. brevicornu (746.71 U/mL), E. pubescens (728.18 U/mL) and E. acuminatum (528.87 U/mL), which indicated that E. sagittatum had a good anti-inflammatory biological potency, and the counterfeit E. acuminatum had lower anti-inflammatory potency than the four genuine ones. Correlation analysis showed there was a significant positive correlation between icariin and the anti-inflammatory potency of Epimedii Folium (r = 0.540, P < 0.05), suggesting that this component may play a major role in the anti-inflammatory activity. Conclusion A method for evaluating the quality of Epimedii Folium based on its anti-inflammatory biological potency was established, which can directly reflect the quality of different Epimedii Folium in terms of anti-inflammatory “superiority or inferiority”, and provide a reference for the quality evaluation of traditional Chinese medicines with anti-inflammatory properties.

R285.5

中央本级重大增减支项目“名贵中药资源可持续利用能力建设项目（2060302）；国家自然科学基金重点项目（U23A20519）