基于高通量测序技术探讨健脾化痰方对ApoE^(-/-)AS小鼠VSMC增殖相关miRNA的调控作用  

Study on the Jianpi Huatan Formula Regulating VSMC Proliferation-related miRNA in ApoE^(-/-)AS mice Based on High-throughput Sequencing Technology

作　　者：田苗[1,2] 贾连群[2] 具星 关乐[1] 王洋[1] 闵冬雨[1] 杜高乐 佟惊宇 杨关林[2] Tian Miao;Jia Lianqun;Ju Xing;Guan Le;Wang Yang;Min Dongyu;Du Gaole;Tong Jingyu;Yang Guanlin(Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,China;The First Clinical College of Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China)

机构地区：[1]辽宁中医药大学附属医院,沈阳110032 [2]辽宁中医药大学第一临床学院,沈阳110847

出　　处：《世界科学技术-中医药现代化》2023年第11期3646-3653,共8页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：辽宁省教育厅重点攻关项目(L202057):基于“脾主肌肉”理论探讨健脾祛痰复方通过AMPK通路影响血管平滑肌细胞迁移对动脉粥样硬化的作用,负责人:田苗;沈阳市科学技术局常见多发病医疗技术提升项目(21-173-9-53):健脾祛痰法调控免疫细胞平衡改善动脉粥样硬化的研究,负责人:田苗;国家自然科学基金委员会青年项目(81904079):健脾祛痰方调控miRNA27-PPARγ轴影响AS大鼠血管周围脂肪细胞分化的机制研究,负责人:王洋

摘　　要：目的本研究运用高通量测序技术检测健脾化痰方调控ApoE^(-/-)小鼠血管平滑肌细胞增殖和迁移相关miRNA,并验证相关蛋白的表达,探讨该方对动脉粥样硬化的治疗作用。方法10只C57BL/6J小鼠作为对照组,30只ApoE^(-/-)小鼠随机分为模型组、西药组和健脾化痰中药组,对各组小鼠主动脉样本进行miRNA测序,比较差异表达miRNA,筛选出与血管平滑肌细胞增殖和迁移相关miRNA,使用RT-qPCR和Western blot技术对差异miRNA及相关蛋白进行验证。结果与模型组相比,中药组共有9个miRNA表达发生改变,其中7个上调,2个下调。miRNA-34a在中药组表达上调,经RT-qPCR及Western blot实验验证,中药组小鼠主动脉miRNA-34a及α-SMA蛋白表达与模型组比较均有显著差异(P<0.01)。结论健脾化痰方可能通过调控miRNA-34a抑制血管平滑肌细胞增殖改善ApoE^(-/-)小鼠动脉粥样硬化。Objective This study used high-throughput sequencing technology to detect the regulation of JianPiHuaTan Prescription(JPHT)on the ApoE^(-/-)mouse miRNA related to vascular smooth muscle cells proliferation and migration,verify the expression of related proteins,and explore the therapeutic effect of JPHT on atherosclerosis.Methods Ten of C57BL/6J mice were used as control group,and 30 ApoE^(-/-)mice were randomly divided into model group,western medicine group and JPHT group.Sequencing technology was performed on aortic sample to select the differentially expressed miRNA.We screened out the tagetted miRNA related to VSMCs proliferation and migration.RT-qPCR and Western blot technology were used to test the differentially expressed miRNA and tagetted protein.Results Compared with model group,a total of 9 miRNAs were changed in JPHT group,among which 7 were up-regulated and 2 were down-regulated.The expression of miRNA-34a was up-regulated in JPHT group.The miRNA-34a andα-SMA protein expression in aorta of JPHT group were significantly different with model group by RT-qPCR and Western blot experiment(P<0.01).Conclusion Jianpi Huatan Prescription may improve ApoE^(-/-)mice atherosclerosis through inhibitting the proliferation of vascular smooth muscle cells by regulating miRNA-34a.

关 键 词：miRNA测序 健脾化痰 血管平滑肌细胞 动脉粥样硬化 脾主肌肉 

分 类 号：R285.5[医药卫生—中药学]