目的 利用网络靶点驱动的策略结合动物实验验证探讨丹参关键活性成分群改善血瘀证的作用机制。方法 采用网络药理学技术整合蛋白质-蛋白质相互作用（PPI）网络和京都基因与基因组百科全书（KEGG）通路数据库，预测课题组前期筛选的丹参关键活性成分群改善血瘀证的潜在作用靶点，并通过分子对接技术进行药物分子-靶点相互作用分析；建立盐酸肾上腺素诱导的大鼠急性血瘀证模型，采用ELISA试剂盒检测大鼠血清炎症因子[肿瘤坏死因子（TNF-α）、白细胞介素（IL）-6）]及心肌酶[肌酸激酶同工酶（CK-MB）、乳酸脱氢酶A (LDHA)]水平，利用实时荧光定量PCR (qRT-PCR)和Western blotting对网络预测的丹参关键活性成分群相关的信号通路进行验证。结果 网络靶点预测发现丹参关键活性成分群可能通过作用于SRC、STAT3、TP53、TNF、EGFR、MMP9、BCL2、MYC等核心靶点发挥作用；与对照组相比，模型组Caspase-3蛋白水平明显升高，Bcl-2、p-Akt、p-PI3K和PI3K蛋白水平明显降低（P<0.05）;与模型组相比，治疗组Caspase-3蛋白水平明显降低，Bcl-2、p-Akt和p-PI3K蛋白水平明显升高（P<0.05）。结果表明丹参关键活性组分群可激活PI3K/AKT信号通路，下调Caspase-3蛋白水平并上调Bcl-2蛋白水平来抗心肌细胞凋亡发挥改善血瘀证的功效。结论 丹参关键活性成分群改善血瘀证与激活PI3K/Akt信号通路有关，为丹参的现代药物制剂研发提供实验依据。

Objective To explore the mechanism of the key active component groups of Salvia miltiorrhiza in improving blood stasis syndrome by using a network target-driven strategy combined with animal verification experiments. Methods Network pharmacology was employed to integrate protein-protein interaction（PPI） networks and KEGG pathway databases to predict potential targets of the previously screened key active component groups of S. miltiorrhiza in in improving blood stasis syndrome. Molecular docking technology was used for the analysis of the interaction between drug molecules and targets. A rat model of acute blood stasis syndrome induced by adrenaline hydrochloride was established. ELISA kits were used to detect the levels of serum inflammatory factors（TNF-α, IL-6） and myocardial enzymes（CK-MB, LDHA） in rats. qRT-PCR and Western blotting were utilized to verify the predicted signaling pathways related to the key active component groups of S. miltiorrhiza. Results Network target prediction revealed that “the key active component groups” of S. miltiorrhiza might exert their effects by acting on the core targets such as SRC, STAT3, TP53, TNF, EGFR, MMP9, BCL2, and MYC. Compared with the control group, the protein level of Caspase-3 in the model group was significantly increased, while the protein levels of Bcl-2, p-Akt, p-PI3K and PI3K were significantly decreased（P ＜ 0.05）; Compared with the model group, the protein level of Caspase-3 in the treatment group was significantly decreased, and the protein levels of Bcl-2, p-Akt and p-PI3K were significantly increased（P ＜ 0.05）. These results indicated that the key active component group of S. miltiorrhiza could activate the PI3K/AKT signaling pathway, downregulate the protein level of Caspase-3 and upregulate the protein level of Bcl-2 to inhibit cardiomyocyte apoptosis, thereby improving blood stasis syndrome. Conclusion This study indicates that the improvement of blood stasis syndrome by the key active component groups of S. miltiorrhiza is related to the activation of the PI3K/Akt signaling pathway, providing experimental evidence for the development of modern drug formulations of S. miltiorrhiza.

R285.5

山东中医药学会临床药学科研专项基金项目(SDACM202204); 山东省医药卫生科技项目(202413010355); 国家中医药管理局监测统计中心中医药监测统计研究课题(2025JCTJC165)