目的 基于网络药理学和分子对接分析丹参酮Ⅰ治疗心肌损伤的作用机制,并进行实验验证。方法 运用网络药理学选取丹参酮Ⅰ治疗心肌损伤的关键靶点,利用分子对接预测潜在核心蛋白,并进一步探究丹参酮Ⅰ药效作用,验证其作用机制。以300μmol/L H₂O₂作用HL-1细胞建立细胞损伤模型,模型建立后将细胞分为对照组、模型组、卡托普利组及丹参酮Ⅰ 0.1、1、10μmol/L组。给药干预24 h后用CCK-8法和LDH法检测细胞活力,F-actin法检测细胞骨架损伤情况,Elisa法检测细胞损伤后炎症因子表达。最后通过Western blotting验证其对核心蛋白的影响。结果 网络药理学预测显示,丹参酮Ⅰ治疗心肌损伤交集靶点72个。通过PPI网络筛选得到丹参酮Ⅰ对治疗心肌损失的关键治疗靶点7个,即表皮生长因子受体(EGFR)、前列腺素内过氧化物合酶2(PTGS2)、丝裂原活化蛋白激酶14(MAPK14)、蛋白酪氨酸磷酸酶受体C(PTPRC)、基质金属蛋白酶2(MMP2)、血管内皮生长因子受体2(KDR)、沉默调节蛋白1(SIRT1)。分子对接筛选出丹参酮Ⅰ治疗心肌损伤核心靶蛋白主要是PTGS2、MMP2和MAPK14。细胞药效结果显示,丹参酮Ⅰ能显著抑制细胞凋亡率,改善细胞生长状况及细胞损伤情况,调控细胞损伤后炎症因子白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)水平。Western blotting结果进一步证实丹参酮Ⅰ通过对核心蛋白PTGS2、MMP2、MAPK14的抑制发挥调控作用。结论 丹参酮Ⅰ能有效抑制H₂O₂诱导的HL-1细胞损伤,其机制可能与调控PTGS2、MMP2和MAPK14蛋白的表达有关。

Objective The mechanism of tanshinone Ⅰ in treatment of myocardial injury was analyzed based on network pharmacology and molecular docking, and experimental verification was conducted.Methods Network pharmacology was used to select the key targets of tanshinone Ⅰ in treatment of myocardial injury, and molecular docking was used to predict potential core proteins, and further explore the efficacy of tanshinone I and verify its mechanism of action. HL-1 cells were treated with 300 μmol/L H₂O₂ to establish the cell damage model. After the model was established, the cells were divided into control group, model group, captopril group, tanshinone I 0.1, 1, 10 μmol/L groups. After 24 h of intervention, CCK-8 method and LDH method were used to detect cell viability, F-actin method was used to detect cytoskeletal injury, and Elisa method was used to detect the expression of inflammatory factors after cell injury. Finally, Western blotting was used to verify its effect on core proteins.Results Network pharmacologic prediction showed that tanshinone I treated 72 intersection targets of myocardial injury. Seven key therapeutic targets of tanshinone I for the treatment of myocardial loss were identified through PPI network screening. Epidermal growth factor receptor(EGFR), prostaglandin endoperoxidase synthase 2(PTGS2), mitogen activated protein kinase 14(MAPK14), protein tyrosine phosphatase receptor C(PTPRC), MMP2, vascular endothelial growth factor receptor 2(KDR), silencing regulatory protein 1(SIRT1). PTGS2, MMP2 and MAPK14 were the core target proteins of tanshinone I in treatment of myocardial injury. The results of cell efficacy showed that tanshinone I could significantly inhibit cell apoptosis rate, improve cell growth and cell damage, regulate the expression of inflammatory cytokines interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) after cell injury. Western blotting results further confirmed that tanshinone I played a regulatory role by inhibiting core proteins PTGS2, MMP2 and MAPK14.Conclusion Tanshinone I can effectively inhibit H₂O₂-induced HL-1 cell damage, and its mechanism may be related to the regulation of PTGS2,MMP2 and MAPK14 protein expression.

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国家重点研发计划项目(2022YFC3500300)