目的 基于網(wǎng)絡(luò)藥理學(xué)和分子對接分析丹參酮Ⅰ治療心肌損傷的作用機制,并進行實驗驗證。方法 運用網(wǎng)絡(luò)藥理學(xué)選取丹參酮Ⅰ治療心肌損傷的關(guān)鍵靶點,利用分子對接預(yù)測潛在核心蛋白,并進一步探究丹參酮Ⅰ藥效作用,驗證其作用機制。以300μmol/L H₂O₂作用HL-1細胞建立細胞損傷模型,模型建立后將細胞分為對照組、模型組、卡托普利組及丹參酮Ⅰ 0.1、1、10μmol/L組。給藥干預(yù)24 h后用CCK-8法和LDH法檢測細胞活力,F-actin法檢測細胞骨架損傷情況,Elisa法檢測細胞損傷后炎癥因子表達。最后通過Western blotting驗證其對核心蛋白的影響。結(jié)果 網(wǎng)絡(luò)藥理學(xué)預(yù)測顯示,丹參酮Ⅰ治療心肌損傷交集靶點72個。通過PPI網(wǎng)絡(luò)篩選得到丹參酮Ⅰ對治療心肌損失的關(guān)鍵治療靶點7個,即表皮生長因子受體(EGFR)、前列腺素內(nèi)過氧化物合酶2(PTGS2)、絲裂原活化蛋白激酶14(MAPK14)、蛋白酪氨酸磷酸酶受體C(PTPRC)、基質(zhì)金屬蛋白酶2(MMP2)、血管內(nèi)皮生長因子受體2(KDR)、沉默調(diào)節(jié)蛋白1(SIRT1)。分子對接篩選出丹參酮Ⅰ治療心肌損傷核心靶蛋白主要是PTGS2、MMP2和MAPK14。細胞藥效結(jié)果顯示,丹參酮Ⅰ能顯著抑制細胞凋亡率,改善細胞生長狀況及細胞損傷情況,調(diào)控細胞損傷后炎癥因子白細胞介素-6(IL-6)、腫瘤壞死因子-α(TNF-α)水平。Western blotting結(jié)果進一步證實丹參酮Ⅰ通過對核心蛋白PTGS2、MMP2、MAPK14的抑制發(fā)揮調(diào)控作用。結(jié)論 丹參酮Ⅰ能有效抑制H₂O₂誘導(dǎo)的HL-1細胞損傷,其機制可能與調(diào)控PTGS2、MMP2和MAPK14蛋白的表達有關(guān)。

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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國家重點研發(fā)計劃項目(2022YFC3500300)