红参调控DARS-AS1/miR-125a-5p通路对人血管平滑肌细胞生物学过程的影响

Effects of Red Ginseng on Biological Processes of Human Vascular Smooth Muscle Cells by Regulating DARS-AS1/miR-125a-5p Pathway

目的探讨红参是否通过调控DARS-AS1/miR-125a-5p通路从而抑制人血管平滑肌细胞增殖、迁移及侵袭。方法体外培养人血管平滑肌细胞,同型半胱氨酸(Hcy)处理细胞建立模型,使用不同剂量的红参处理细胞;四唑盐(MTT)进行血管平滑肌细胞增殖检测;Transwell小室实验进行迁移及侵袭检测;实时荧光定量聚合酶链式反应(qRT-PCR)检测DARS-AS1、miR-125a-5p的表达量;Hcy诱导的细胞中分别转染si-DARS-AS1、pcDNA-DARS-AS1,检测细胞增殖、迁移及侵袭;双荧光素酶报告实验检测DARS-AS1、miR-125a-5p的靶向关系。结果Hcy可增高血管平滑肌细胞增殖率、迁移细胞数、侵袭细胞数,促进DARS-AS1表达,抑制miR-125a-5p表达(P<0.05);红参可降低细胞增殖率,减少迁移细胞数、侵袭细胞数,抑制DARS-AS1表达,促进miR-125a-5p表达(P<0.05),且呈剂量依赖性;干扰DARS-AS1表达可抑制Hcy诱导的人血管平滑肌细胞增殖、迁移及侵袭;双荧光素酶报告实验证实DARS-AS1可靶向结合miR-125a-5p;DARS-AS1过表达以减弱红参对Hcy诱导的人血管平滑肌细胞增殖、迁移及侵袭的抑制作用。结论红参可能通过调控DARS-AS1/miR-125a-5p通路从而抑制血管平滑肌细胞增殖、迁移及侵袭。

Objective To investigate whether red ginseng inhibits the proliferation,migration and invasion of human vascular smooth muscle cells by regulating the DARS-AS1/miR-125A-5P pathway.Methods Human vascular smooth muscle cells were cultured in vitro and treated with homocysteine(Hcy)to establish the model.The proliferation of vascular smooth muscle cells was detected by tetrazolium salt(MTT).Transwell chamber assay was used to detect migration and invasion.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression of DARS-AS1 and miR-125a-5p.The Hcy induced cells were transfected with si-DARS-AS1 and pcDNA-DARS-AS1,respectively,and the proliferation,migration and invasion of the cells were detected by the above methods.Dual luciferase reporter assay was used to detect the targeting relationship between DARS-AS1 and miR-125A-5p.Results Hcy could increase the proliferation rate,the number of migrating cells,and the number of invading cells,promote the expression of DARS-AS1,and inhibit the expression of miR-125a-5p(P<0.05).Red ginseng could reduce the cell proliferation rate,reduce the number of migrating cells and invading cells,inhibit the expression of DARS-AS1,and promote the expression of miR-125a-5p(P<0.05)in a dose-dependent manner.Interference of DARS-AS1 expression could inhibit Hcy induced proliferation,migration and invasion of human vascular smooth muscle cells.Dual luciferase reporter assay confirmed that DARS-AS1 could target miR-125A-5p.DARS-AS1 overexpression could attenuate the inhibitory effect of red ginseng on Hcy induced proliferation,migration and invasion of human vascular smooth muscle cells.Conclusion Red ginseng may inhibit the proliferation,migration and invasion of vascular smooth muscle cells by regulating the DARS-AS1/miR-125a-5P pathway.