摘要
目的研究苏木乙酸乙酯提取液(SAEE)对氧化型低密度脂蛋白(ox-LDL)诱导的血管平滑肌细胞(VSMC)增殖、凋亡的机制及miRNA-99a在其中的可能作用。方法 ox-LDL诱导VSMC模型,分别将SAEE(200μg/mL)、miRNA-99a模拟剂、miRNA-99a抑制剂作用于细胞。采用MTT法检测各组VSMC的增殖,流式细胞学检测其凋亡水平,Western blotting法检测细胞中Ki-67、PCNA、Bax、Bcl-2、MMP-1、TIMP-1蛋白的表达,RT-PCR检测细胞中miRNA-99a的表达。结果与空白组相比,ox-LDL能显著增加VSMC细胞增殖、晚期凋亡和总凋亡率,上调细胞中Ki-67、PCNA、Bax、MMP-1蛋白的表达及Bax/Bcl-2、MMP-1/TIMP-1的比值,下调Bcl-2、TIMP-1蛋白和miRNA-99a的表达,差异均具有显著统计学意义(P <0.01);SAEE能够抑制VSMC细胞增殖、晚期凋亡和总凋亡率,上调Bcl-2、TIMP-1蛋白和mi R-99a的表达,下调细胞中Ki-67、PCNA、Bax、MMP-1蛋白的表达及Bax/Bcl-2、MMP-1/TIMP-1的比值,与模型组比较,差异均具有统计学意义(P <0.01)。结论 SAEE能够通过上调细胞中miRNA-99a的表达,抑制VSMC细胞增殖、凋亡和细胞外基质的降解,促进斑块的稳定和防止纤维帽的破裂,发挥抗动脉粥样硬化的作用。
Objective: To study the mechanism of SAEE on proliferation and apoptosis of VSMC induced by ox-LDL and the possible role of miRNA-99a in atherosclerosis. Methods: VSMC model was induced by ox-LDL. SAEE(200 μg/mL),miR-99a mimics and miR-99a inhibitor were used to treat the cells respectively. MTT method was used to detect the proliferation of VSMC,flow cytometry was used to detect cell apoptosis;Western-blot method was used to detect the expression of Ki-67,PCNA,Bax,Bcl-2,MMP-1 and TIMP-1 protein;RT-PCR was used to detect the expression of microRNA-99a. Results: Compared with the control group,ox-LDL could significantly increase the proliferation,late apoptosis and total apoptosis rate of VSMC cells,up-regulate the expression of Ki-67,PCNA,Bax,MMP-1 protein and the ratio of Bax/Bcl-2,MMP-1/TIMP-1,down-regulate the expression of Bcl-2,TIMP-1 protein and miRNA-99a(P < 0.01). SAEE could inhibit the proliferation,late apoptosis and total apoptosis rate of VSMC cells,up-regulate the expression of Bcl-2,TIMP-1 protein and miRNA-99a,down-regulate the expression of Ki-67,PCNA,Bax,MMP-1 protein and the ratio of Bax/Bcl-2 and MMP-1/TIMP-1 in VSMC cells,which had significant difference when compared with the model group(P < 0.01). Conclusion: SAEE can inhibit the proliferation,apoptosis and ECM degradation of VSMC cells by up-regulating the expression of miRNA-99a,promote the stability of plaque and prevent the rupture of fibrous cap,and play an anti-atherosclerosis role.
作者
于玲
曹丽娟
孙静
周亚滨
Yu Ling;Cao Lijuan;Sun Jing(Heilongjiang University of Chinese Medicine,Harbin 150040,China)
出处
《中国中医急症》
2019年第4期585-589,共5页
Journal of Emergency in Traditional Chinese Medicine
基金
国家科技重大专项课题(2009ZX09103-425)
黑龙江省科技攻关计划项目(GC06C32804)
黑龙江省教育厅面上项目(11511375)
黑龙江省普通高等学校青年学术骨干支持计划(1253G055)
