目的 基于网络药理学和分子对接技术筛选补肾活血方防治糖尿病性视网膜病变的质量评价指标，同时 建立活性成分的超高效液相色谱-三重四极杆质谱（UHPLC-QqQ-MS/MS）含量测定方法。方法 利用网络药理 学筛选补肾活血方防治糖尿病性视网膜病变的关键化合物和靶点。基于分子对接技术验证化合物和靶点的相关 性，确定补肾活血方的活性成分作为质量评价指标。建立补肾活血方活性成分的 UHPLC-QqQ-MS/MS 含量测 定方法。采用 ZORBAX SB-C18 （2.1 mm × 50 mm，1.8 μm）色谱柱，甲醇（A）-0.1% 甲酸水溶液（B）作为流动相， 在多反应监测模式下进行梯度洗脱。流速：0.3 mL·min-1；进样量：1 μL。结果 网络药理学及分子对接筛选 出了 5 个核心靶点及 12 个活性成分（毛蕊花糖苷，松果菊苷，异类叶升麻苷，丹参酮ⅡA，隐丹参酮，二氢丹 参酮 I，人参皂苷 Re、Rd 和 Rb1，葛根素，大豆苷，鹰嘴豆牙素 A），其亲和力良好（结合能≤ -5.0 kcal·mol-1）。 含量测定结果表明，各成分在线性范围内相关性良好（r＞0.999 5），平均加样回收率在 97.57%～101.48%。 8 批样品中 12 个成分含量分别为 0.027 9%～0.050 6%、0.006 4%～0.022 0%、0.017 1%～0.041 5%、0.009 2%～ 0.015 4%、 0.012 6%～0.020 5%、 0.004 4%～0.007 6%、 0.334%～0.643%、 0.238%～0.530%、 0.353%～ 0.693%、3.411%～6.048%、1.023%～1.352% 和 0.000 8%～0.001 8%。结论 基于网络药理学、分子对接和 UHPLC-QqQ-MS/MS 技术，建立了快速筛选并测定补肾活血方中 12 个活性成分的方法，为全面评价其质量及 有效性提供了参考。

Objective To screen the quality evaluation indicators of Bushen Huoxue Prescription （BHP） in the treatment of diabetic retinopathy （DR） by network pharmacology and molecular docking technology， and to establish content determination of active components in BHP by ultra-high-performance liquid chromatography triplequadrupole mass spectrometry （UHPLC-QqQ-MS/MS） . Methods Network pharmacology was used to screen the disease-related targets and key components，followed by molecular docking to further verify the interaction between them and confirm the active ingredients in BHP as the quality evaluation indicators. A UHPLC-QqQ-MS/MS method for the content determination of the active ingredients in BHP was established. A ZORBAX SB-C18 column（2.1 mm × 50 mm，1.8 μm）was used，and methanol （A）-0.1% formic acid solution （B）was used as mobile phase. Gradient elution was performed in multiple reaction monitor mode. The flow rate was 0.3 mL·min-1 and the injection volume was 1 μL. Results Network pharmacology and molecular docking revealed five core targets and 12 BHP-related components （verbascoside， echinacoside， isoacteoside， tanshinone IIA， cryptotanshinone， dihydrotanshinone I， ginsenoside Re， Rd and Rb1， puerarin， daidzin， biochanin A） for the treatment of DR. There was a strong binding affinity between them （binding energy ≤ -5.0 kcal·mol-1）. The established quantitative method demonstrated each component presented a good linearity within the specified range （r＞0.999 5） . The average recovery was in the range of 97.57%～101.48%. The contents of 12 components in eight batches of BHP samples were 0.027 9%～ 0.050 6%， 0.006 4%～0.022 0%， 0.017 1%～0.041 5%， 0.009 2%～0.015 4%， 0.012 6%～0.020 5%， 0.004 4%～0.007 6%， 0.334%～0.643%， 0.238%～0.530%， 0.353%～0.693%， 3.411%～6.048%， 1.023%～ 1.352%， 0.000 8%～0.001 8%， respectively. Conclusion Based on network pharmacology， molecular docking and UHPLC-QqQ-MS/MS，a method for rapid screening and determination of 12 active components of BHP in the prevention and treatment of DR was established. This study provided a reference for comprehensive assessment of the quality and effectiveness of BHP.

R285.5；R284.1

国家自然科学基金面上项目（82174273，81774202，82305323）