目的 探讨中药复方血管软化丸通过miRNA-467b靶向脂蛋白脂肪酶（lipoprotein lipase，LPL）调控巨噬细胞，减少白细胞介素（interleukin，IL）-6，IL-1β、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）和单核细胞趋化蛋白-1（monocyte chemoattractant protein-1，MCP-1）等炎症因子分泌和巨噬细胞脂质蓄积，从而发挥抗动脉粥样硬化的作用与初步机制。方法 将RAW264.7巨噬细胞随机分为5组，即对照组、模型组、miR-467b mimic组、中药高剂量含药血清组、中药低剂量含药血清组；经干预后，采用RT-PCR检测巨噬细胞中pri-miR-467b和LPL mRNA表达；采用高效液相色谱法检测巨噬细胞内胆固醇水平。连续4周高脂饲料（含脂肪21%、胆固醇0.15%）饲养ApoE-/-小鼠复制动脉粥样硬化模型，模型复制成功后，中药高、低剂量组每日分别灌胃血管软化丸86.4、21.6 g/kg，连续给药12周；对照组、模型组每日灌胃0.9%生理盐水86.4 g/kg 。采用免疫组织化学法检测主动脉LPL蛋白表达水平，RT-PCR检测主动脉pri-miR-467b和LPL mRNA表达水平；采用ELISA法检测血清IL-6、IL-1β、TNF-α、MCP-1水平。结果 与对照组比较，模型组巨噬细胞pri-miR-467b mRNA表达水平和巨噬细胞内游离胆固醇（free cholesterol，FC）水平均明显降低（P＜0.05），巨噬细胞内LPL mRNA及其蛋白表达水平以及总胆固醇（total cholesterol，TC）、胆固醇酯（cholesterol ester，CE）水平均明显升高（P＜0.05）。血管软化丸能够无剂量依赖性地升高巨噬细胞pri-miR-467b mRNA表达水平和巨噬细胞内FC水平（P＜0.05），降低巨噬细胞中LPL mRNA及其蛋白表达水平以及TC、CE水平（P＜0.05）。血管软化丸能够升高主动脉pri-miR-467b mRNA表达水平（P＜0.05），降低主动脉中LPL mRNA及其蛋白表达水平以及血清中IL-6、IL-1β、TNF-α、 MCP-1水平（P＜0.05）。结论 血管软化丸抑制动脉粥样硬化斑块形成的作用机制可能与调控miRNA-467b靶向LPL调控巨噬细胞，减少IL-6、IL-1β、TNF-α、MCP-1等炎症因子分泌和巨噬细胞脂质蓄积相关。
Objective To investigate the anti-atherosclerotic role of Xueguan Ruanhua Pill and its preliminary mechanism by targeting lipoprotein lipase (LPL) to regulate macrophages through miRNA-467b and reducing the secretion of inflammatory factors such as interleukin-6 (IL-6), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and monocyte chemoattractant protein 1 (MCP-1) and lipid accumulation in macrophages.Methods RAW264.7 macrophages were randomly divided into control group, model group, miR-467b mimic group, high-dose drug-containing serum group, and low-dose drug-containing serum group. After intervention, RT-PCR was used to measure the mRNA expression of pri-miR-467b and LPL in macrophages, and high-performance liquid chromatography was used to measure the level of cholesterol in macrophages. ApoE-/- mice were fed with high-fat diet (containing 21% fat and 0.15% cholesterol) for four consecutive weeks to establish a model of atherosclerosis; after the model was successfully established, the mice in the high- and low-dose Xueguan Ruanhua Pill groups were given Xueguan Ruanhua Pill at a dose of 86.4 and 21.6 g/kg, respectively, by gavage daily for 12 consecutive weeks, and those in the control group and the model group were given 0.9% normal saline at a dose of 86.4 g/kg by gavage daily. Immunohistochemistry was used to measure the protein expression of LPL in the aorta, RT-PCR was used to measure the mRNA expression of pri-miR-467b and LPL in the aorta, and ELISA was used to measure the serum levels of IL-6, IL-1β, TNF-α, and MCP-1. Results Compared with the control group, the model group had significant reductions in the mRNA expression of pri-miR-467b and the level of free cholesterol (FC) in macrophages (P<0.05) and significant increases in the mRNA and protein expression of LPL and the levels of total cholesterol (TC) and cholesteryl ester (CE) in macrophages (P<0.05). Xueguan Ruanhua Pill significantly increased the mRNA expression of pri-miR-467b and the level of FC in macrophages (P<0.05) and reduced the mRNA and protein expression of LPL and the levels of TC and CE in macrophages (P<0.05), without a dose-dependent manner. Xueguan Ruanhua Pill significantly increased the mRNA expression of pri-miR-467b in the aorta (P<0.05) and significantly reduced the mRNA and protein expression of LPL in the aorta and the serum levels of IL-6, IL-1β, TNF-α, and MCP-1 (P<0.05). Conclusion Xueguan Ruanhua Pill can inhibit the formation of atherosclerotic plaque, possibly by targeting LPL to regulate macrophages through miRNA-467b and reducing the secretion of inflammatory factors such as IL-6, IL-1β, TNF-α, and MCP-1 and lipid accumulation in macrophages.