目的 基于人脐静脉内皮细胞（HUVECs）外泌体 miR-145 调控人主动脉血管平滑肌细胞（HAVSMCs）表 型转化角度探讨温心方治疗冠心病的作用机制。方法 给予大鼠温心方水煎剂 33.75 g·kg-1灌胃，每天 1 次， 连续 7 d ，制备含药血清。（1）取对数生长期的 HUVECs，分别设立模型组、温心方组、miR-145 mimic 组、 miR-145 inhibitor 组、miR-145 inhibitor+温心方组。模型组加入 50 mg·L-1 ox-LDL 培养 24 h；温心方组加入 25% 温心方含药血清培养 24 h 后，再加入 ox-LDL 培养 24 h；miR-145 mimic 组、miR-145 inhibitor 组分别用 miR-145 过表达质粒及小干扰 RNA 转染后，再加入 ox-LDL 培养 24 h；miR-145 inhibitor+温心方组在转染后 加入 25% 温心方含药血清培养 24 h，再加入 50 mg·L-1 ox-LDL 培养 24 h。采用高速离心分离获取 HUVECs 外 泌体，电镜观察外泌体形态，BCA 法及 NTA 法对外泌体蛋白浓度及粒径进行测定。（2）取对数生长期 HAVSMCs，分别设立对照组、模型外泌体组、温心方外泌体组、miR-145 mimic 外泌体组、miR-145 inhibitor 外泌体组、miR-145 inhibitor+温心方外泌体组、Wnt 抑制剂组。对照组继续使用培养基培养；Wnt 抑制剂组加 入 50 ng XAV-939（10 μmol·L-1 ）；其余各组加入对应外泌体 50 ng（外泌体终浓度为 100 μg·mL-1 ），培养 24 h。 采用 MTT 法检测 HAVSMCs 细胞增殖率；Transwell 法检测细胞迁移能力；流式细胞术检测细胞凋亡及细胞周 期；qRT-PCR 法检测细胞 miR-145、WNT2B 基因表达水平；Western Blot 法检测细胞 α 平滑肌肌动蛋白 （α-SMA）、平滑肌 22α 蛋白（SM22α）、骨桥蛋白（OPN）、Wnt1、β-catenin、LRP6、GSK-3β 蛋白表达水平。 结果 成功提取 HUVECs 外泌体，平均粒径为 140.6 nm，浓度为 1.6×108个/mL。与对照组比较，模型外泌体 组的 HAVSMCs 细胞增殖率明显升高（P＜0.05），细胞迁移数显著增加（P＜0.01）；细胞凋亡率显著升高（P＜ 0.01），S 期细胞占比明显增加（P＜0.05）；HAVSMCs miR-145 基因表达显著下调（P＜0.01），WNT2B 基因表达 显著上调（P＜0.01）；HAVSMCs 收缩型标志蛋白 α-SMA、SM22α 表达显著下调（P＜0.01），分泌型标志蛋白 OPN 及 Wnt/β-catenin 信号通路关键蛋白 Wnt1、β-catenin、LRP6、GSK-3β 表达显著上调（P＜0.01）。与模型 外泌体组比较，温心方外泌体组的 HAVSMCs 细胞增殖率明显降低（P＜0.05），miR-145 inhibitor 外泌体组的细 胞增殖率明显升高（P＜0.05）；温心方外泌体组、miR-145 mimic 外泌体组的 HAVSMCs 细胞迁移数明显减少 （P＜0.05），miR-145 inhibitor 外泌体组的细胞迁移数明显增加（P＜0.05）；温心方外泌体组、Wnt 抑制剂组的 S 期细胞明显减少（P＜0.05）；温心方外泌体组、miR-145 mimic 外泌体组 HAVSMCs miR-145 基因表达明显上 调（P＜0.05），WNT2B 基因表达明显下调（P＜0.05）；温心方外泌体组、miR-145 mimic 外泌体组、Wnt 抑制剂 组 HAVSMCs 的 α-SMA、SM22α 蛋白表达明显上调（P＜0.05），OPN、Wnt1、β-catenin、LRP6、GSK-3β 蛋白 表达明显下调（P＜0.05）；miR-145 inhibitor 外泌体组 HAVSMCs 的 α-SMA、SM22α 蛋白表达明显下调（P＜0.05），OPN 蛋白表达明显上调（P＜0.05）。与 miR-145 inhibitor 外泌体组比较，miR-145 inhibitor+温心方外泌 体组、Wnt 抑制剂组的 HAVSMCs 细胞增殖率明显降低（P＜0.05），细胞迁移数明显减少（P＜0.05）；miR-145 inhibitor+温心方外泌体组 HAVSMCs 的 α-SMA、SM22α 蛋白表达明显上调（P＜0.05），OPN、Wnt1、β-catenin、 LRP6、GSK-3β 蛋白表达明显下调（P＜0.05）。结论 温心方干预的 HUVECs 外泌体可上调 HAVSMCs 中 miR-145 表达，通过靶向 WNT2B 及抑制 Wnt/β-catenin 信号通路调控 HAVSMCs 向收缩型转化，抑制其增殖、迁移，减 轻血管内膜增生，从而发挥防治冠状动脉粥样硬化的作用。

Objective To explore the mechanism of Wenxin Formula in the treatment of coronary heart disease from the perspective of regulating human umbilical vein endothelial cell （HUVECs） exosome miR-145 expression to inhibit phenotype transformation of human aortic vascular smooth muscle cells （HAVSMCs）. Methods Rats were administered Wenxin Formula decoction （33.75 g · kg-1 ） by gavage once daily for 7 days to prepare drug-containing serum. （1）HUVECs in the logarithmic growth phase were divided into model，Wenxin Formula，miR-145 mimic，miR-145 inhibitor，and miR-145 inhibitor + Wenxin Formula groups. The model group was treated with 50 mg·L-1 ox-LDL for 24 hours； the Wenxin Formula group was treated with 25% Wenxin Formula drug-containing serum for 24 hours， followed by ox-LDL for 24 hours；the miR-145 mimic and miR-145 inhibitor groups were transfected with miR-145 overexpression plasmid or siRNA，respectively，followed by ox-LDL treatment for 24 hours；the miR-145 inhibitor + Wenxin Formula group was transfected and then treated with 25% Wenxin Formula drug-containing serum for 24 hours， followed by 50 mg·L-1 ox-LDL for 24 hours. Exosomes were isolated from HUVECs by ultracentrifugation，and their morphology was observed by electron microscopy. Exosome protein concentration and particle size were measured using BCA and NTA assays.（2）HA-VSMCs in the logarithmic growth phase were divided into control， model exosome， Wenxin Formula exosome， miR-145 mimic exosome， miR-145 inhibitor exosome， miR-145 inhibitor + Wenxin Formula exosome，and Wnt inhibitor groups. The control group was cultured in medium；the Wnt inhibitor group was treated with 50 ng XAV-939（10 μmol·L-1 ）；the other groups were treated with 50 ng of corresponding exosomes （final concentration：100 μg·mL-1 ）for 24 hours. HA-VSMC proliferation was assessed by MTT assay；cell migration was evaluated using Transwell assay；apoptosis and cell cycle were analyzed by flow cytometry；miR-145 and WNT2B gene expression levels were detected by qRT-PCR； and protein expression levels of α -SMA， SM22α， OPN， Wnt1， β-catenin，LRP6，and GSK-3β were measured by Western Blot. Results Exosomes were successfully extracted from HUVECs，with an average particle size of 140.6 nm and a concentration of 1.6×108 particles/mL. Compared with the control group，the model exosome group showed significantly increased HAVSMCs proliferation （P＜0.05），enhanced cell migration （P＜0.01）， elevated apoptosis rate （P＜0.01）， and increased S-phase cell proportion （P＜0.05）. HAVSMCs miR-145 expression was significantly downregulated （P＜0.01），while WNT2B expression was significantly upregulated （P＜0.01）. Contractile markers of HAVSMCs α-SMA and SM22α were significantly downregulated （P＜ 0.01），while secretory marker OPN and Wnt/β-catenin signaling pathway proteins Wnt1，β-catenin，LRP6，and GSK-3β were significantly upregulated （P＜0.01）. Compared with the model exosome group， the Wenxin Formula exosome group showed significantly reduced HAVSMCs proliferation （P＜0.05），while the miR-145 inhibitor exosome group exhibited increased proliferation （P＜0.05）. The Wenxin Formula exosome and miR-145 mimic exosome groups showed significantly reduced HAVSMCs cell migration （P＜0.05）， while the miR-145 inhibitor exosome group exhibited increased migration （P＜0.05）. The Wenxin Formula exosome and Wnt inhibitor groups showed reduced S-phase cell proportion （P＜0.05）. The Wenxin Formula exosome and miR-145 mimic exosome groups showed upregulated HAVSMCs miR-145 expression （P＜0.05） and downregulated WNT2B expression （P＜0.05）. The Wenxin Formula exosome， miR-145 mimic exosome， and Wnt inhibitor groups showed upregulated α -SMA and SM22α protein expression （P＜0.05） and downregulated OPN，Wnt1，β-catenin，LRP6，and GSK-3β protein expression （P＜0.05）. The miR-145 inhibitor exosome group showed downregulated α-SMA and SM22α protein expression （P＜ 0.05） and upregulated OPN expression （P＜0.05）. Compared with the miR-145 inhibitor exosome group，the miR-145 inhibitor + Wenxin Formula exosome and Wnt inhibitor groups showed significantly reduced HAVSMCs proliferation （P＜0.05） and cell migration （P＜0.05）. The miR-145 inhibitor + Wenxin Formula exosome group showed upregulated α-SMA and SM22α protein expression （P＜0.05） and downregulated OPN，Wnt1，β-catenin，LRP6，and GSK-3β protein expression （P＜0.05）. Conclusion Exosomes derived from HUVECs treated with Wenxin Formula upregulate miR-145 expression in HAVSMCs，targeting WNT2B and inhibiting the Wnt/β-catenin signaling pathway to promote HAVSMCs contractile phenotype transformation， inhibit proliferation and migration， and alleviate vascular intimal hyperplasia，thereby exerting preventive and therapeutic effects on coronary atherosclerosis.

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国家自然科学基金项目（82104775）；湖南省科技创新计划项目（2024RC3198）；湖南省自然科学基金项目（2025JJ50567）；湖南省教育厅 科研项目（23B0357）；湖南中医药大学科研项目（Z2023XJYQ04）；中国科学技术学会青年人才托举工程项目（2022QNRC001）。