舒脉胶囊干预血管内皮细胞-平滑肌细胞共培养模型的作用机制研究  

Mechanism of inhibiting the expression of fatty acid binding protein 4 on Shumai Capsule's intervention in the co-culture model of vascular endothelial cells and smooth muscle cells

作　　者：焦琳茜 徐旭英[1] 郭卉 吕春燕 刘威池 丁毅[1] JIAO Lin-xi;XU Xu-ying;GUO Hui;LU Chun-yan;LIU Wei-chi;DING Yi(Beijing Hospital of Traditional Chinese Medicine Affiliated to the Capital Medical University,Beijing 100010,China)

机构地区：[1]首都医科大学附属北京中医医院,北京100010

出　　处：《北京中医药》2023年第7期708-715,共8页Beijing Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(82174388);北京市百千万人才工程培养经费资助(2019A30);第五批全国中医临床优秀人才(国中医药办人教函[2021]271号);北京市医院管理中心“登峰”人才培养计划(DFL20220801)。

摘　　要：目的研究舒脉胶囊对血管内皮细胞(VECs)-血管平滑肌细胞(VSMCs)共培养模型的影响及其可能存在的机制。方法实验一:建立血管内皮细胞-平滑肌细胞(VV)共培养模型,将细胞分为模型组(VV-空白)、5%胎牛血清(FBS)组(VV-5%FBS)、10%FBS组(VV-10%FBS)、5%舒脉胶囊(MS)组(VV-5%MS)、10%MS组(VV-10%MS)5组,采用CCK-8法检测VECs及VSMCs增殖及迁移能力;免疫荧光检测增殖细胞核抗原(PCNA)、一氧化氮合酶(eNOs)、血管内皮生长因子(VEGF)、基质金属蛋白酶2(MMP-2)、α平滑肌肌动蛋白(α-SMA);PCR检测过氧化物酶体增殖物激活受体γ(PPARγ)、脂肪酸结合蛋白(FABP)4基因表达含量;Western Blot检测细胞中PPARγ、FABP4蛋白表达量。实验二:构建FABP4重组质粒,通过质粒转染建立FABP4过表达重组VECs(OF-VECs),及FABP4过表达重组VECs-VSMCs(OFVV)模型,分为模型组(OFVV-空白)、OFVV-MS组、OFVV-抑制剂(InF)组(OFVV-InF)、OFVV-MS+InF组、阳性对照组[OFVV-西洛他唑(Cil)]5组,采用CCK-8法检测OF-VECs及VSMCs增殖及迁移能力;免疫荧光检测PCNA、eNOs、MMP-2、α-SMA;PCR检测PPARγ、FABP4基因表达含量;Western Blot检测细胞中PPARγ、FABP4蛋白表达量。结果实验一:10%MS组、5%MS组VECs的OD值均高于10%FBS组、5%FBS组,VSMCs的OD值均低于10%FBS组、5%FBS组,差异有统计学意义(P<0.01);10%MS组、5%MS组FAPB4、PPARγ基因及蛋白表达量均小于10%FBS组、5%FBS组,差异有统计学意义(P<0.05)。实验二:OFVV-MS+InF组OF-VECs的OD值高于其余各组,VSMCs的OD值低于其余各组,差异有统计学意义(P<0.01);OFVV-MS+InF组FAPB4、PPARγ蛋白表达量均高于其余各组,差异有统计学意义(P<0.001)。结论舒脉胶囊通过调节FABP4表达,增强VECs、降低VSMCs的迁移、增殖能力,改善血管内皮功能,抑制介入术后再狭窄的发生。Objective To study the effect of Shumai Capsule on the co-culture model of vascular endothelial cells(VECs)and vascular smooth muscle cells(VSMCs)and its possible mechanism.Methods In the first experiment,vascular endothelial cells-smooth muscle cells(VV)co-culture model was established,which was divided into five groups:VV-control group,VV-FBS(5%)grouup,VV-FBS(10%)group and VV-Shumai capsule(VV-Medicated serum,VV-MS)(5%)group and VV-Shumai Capsule(10%)group.The proliferation and migration ability of VECs and VSMCs were detected by CCK-8 method.Proliferating cell nuclear antigen(PCNA),nitric oxide synthase(eNOs)and vascular endothelial growth factor(VEGF)were detected by immunofluorescence.The protein expression of PPARγand FABP4 genes were detected by PCR and the gene expression by Western Blot.In the second experiment,the recombinant plasmid FABP4 was constructed,and the overexpressed Fabp4 vascular endothelial cells(of-VECs)were constructed by plasmid transfection,and the co-culture model(OFVV)of FABP4 overexpressed recombinant VECs and VSMCs was established,which was divided into five groups including OFVV-control group,OFVV-MS group,OFVV-InF group,OFVV-MS+InF group,and OFVV-Cil group,CCK-8 method method was used to detect the proliferation and migration ability of OF-VECs and VSMCs.and immunofluorescence to detect PCNA and eNOs.PCR and Western Blot were used to detect the gene and protein expression of PPARγand FABP4 respectively.Results In the first experiment,the OD values of VECs in VV-MS(10%)group and VV-MS(5%)group were all higher than those in VV-FBS(10%)group,while those in VSMCs group were all lower than those in VV-FBS(10%)and VV-FBS(5%)group,and the difference was statistically significant(P<0.01).The expression levels of FAPB4 and PPARγgenes and proteins in VV-MS group were lower than those in VV-FBS group,and the difference was statistically significant(P<0.05).In the second experiment,The OD value of OF-VECs in OFVV-MS+InF group was higher and the OD value of VSMCs was lower than that of othe

关 键 词：舒脉胶囊 脂肪酸结合蛋白4 血管内皮细胞 平滑肌细胞 血管介入术后再狭窄 

分 类 号：R285[医药卫生—中药学]