MiR-665 Regulates Vascular Smooth Muscle Cell Senescence by Interacting With LncRNA GAS5/SDC1.
MiR-665 Regulates Vascular Smooth Muscle Cell Senescence by Interacting With LncRNA GAS5/SDC1.
复制标题
MiR-665 通过与 LncRNA GAS5/SDC1 相互作用调节血管平滑肌细胞衰老
DOI：
10.3389/fcell.2021.700006
复制
发表时间：
2021
影响因子：
5.5
通讯作者：
Zhang L
中科院分区：
文献类型：
--
作者：
Chen T;Liang Q;Xu J;Zhang Y;Zhang Y;Mo L;Zhang L
背景资料：血管衰老被认为是心血管疾病的一个特殊危险因素，而血管平滑肌细胞（vascular smooth muscle cell，VSMCs）在衰老相关的血管重构和动脉粥样硬化的病理过程中起着重要作用。最近的研究报道，长链非编码RNA/microRNA（lncRNA/miRNA）是细胞衰老的关键调节因子。然而，lncRNA GAS 5/miR-665轴在VSMC衰老中的作用和机制仍不完全清楚。研究方法：使用衰老相关的β-gal活性、NAD+/NADH比率和γ H2 AX免疫荧光的免疫荧光染色来评估细胞衰老。通过miRNA微阵列测定鉴定差异表达的miRNA（DEM），随后通过定量实时PCR（qRT-PCR）验证。进行双荧光素酶报告基因测定以确认lncRNA GAS 5和miR-665以及miR-665和多配体蛋白聚糖1（SDC 1）的结合。采用qRT-PCR方法检测93例受试者血清中miR-665、lncRNA GAS 5和SDC 1水平。参与者被分为对照组、老化组和早期血管老化（伊娃）组，并测量他们的臂踝脉搏波速度（baPWV）。结果：通过微阵列分析，在年轻和老年VSMCs中共鉴定出20个重叠的DEM。MiR-665显示出显著的改变，因此选择其用于进一步分析。miR-665在衰老的VSMCs中表达上调，而下调miR-665则可抑制VSMCs的衰老。随后，双荧光素酶报告基因测定确定了miR-665与lncRNA GAS 5和SDC 1的3′-UTR的结合位点。lncRNA GAS 5表达的增加抑制miR-665水平和VSMC衰老。然而，如拯救实验结果所示，miR-665过表达或SDC 1敲低显著逆转了lncRNA GAS 5对VSMC衰老的影响。最后，与对照组相比，衰老组和伊娃组血清中miR-665的表达均较高，尤其是伊娃组。与此相反，两组患者血清lncRNAGAS 5和SDC 1水平均较低。总的来说，在老化和伊娃组中，miR-665表达与lncRNA GAS 5和SDC 1表达呈负相关。结论：miR-665抑制通过负调节SDC 1作为VSMC衰老的重要调节剂发挥作用，SDC 1通过吸收miR-665的lncRNA GAS 5实现。我们的研究结果可能为衰老相关的心血管疾病提供新的治疗策略。
Background: Vascular aging is considered a special risk factor for cardiovascular diseases, and vascular smooth muscle cells (VSMCs) play a major role in aging-related vascular remodeling and in the pathological process of atherosclerosis. Recent research has reported that long non-coding RNA/microRNA (lncRNA/miRNA) is a critical regulator of cellular senescence. However, the role and mechanism of lncRNA GAS5/miR-665 axis in VSMC senescence remain incompletely understood. Methods: Cellular senescence was evaluated using senescence-associated β-gal activity, the NAD+/NADH ratio, and by immunofluorescence staining of γH2AX immunofluorescence. Differentially expressed miRNAs (DEMs) were identified by miRNA microarray assays and subsequently validated by quantitative real-time PCR (qRT-PCR). A dual luciferase reporter assay was conducted to confirm the binding of lncRNA GAS5 and miR-665 as well as miR-665 and syndecan 1 (SDC1). Serum levels of miR-665, lncRNA GAS5, and SDC1 in 93 subjects were detected by qRT-PCR. The participants were subdivided into control, aging, and early vascular aging (EVA) groups, and their brachial-ankle pulse wave velocity (baPWV) was measured. Results: A total of 20 overlapping DEMs were identified in young and old VSMCs via microarray analysis. MiR-665 showed a significant alteration and, therefore, was selected for further analysis. Upregulation of miR-665 was found in aging VSMCs, and downregulation of miR-665 caused an inhibition of VSMCs senescence. Subsequently, the dual luciferase reporter assay determined the binding site of miR-665 with the 3′-UTR of lncRNA GAS5 and SDC1. Increased expression of lncRNA GAS5 expression inhibited the miR-665 level and VSMC senescence. However, as shown in rescue experiment results, either miR-665 overexpression or SDC1 knockdown significantly reversed the effects of lncRNA GAS5 on VSMC senescence. Finally, compared with that of the control group, miR-665 was highly expressed in serum samples in the aging and EVA groups, especially in the EVA groups. On the contrary, serum levels of lncRNA GAS5 and SDC1 were lower in these two groups. Collectively, in the aging and EVA groups, miR-665 expression was negatively correlated with lncRNA GAS5 and SDC1 expression. Conclusion: miR-665 inhibition functions as a vital modulator of VSMC senescence by negatively regulating SDC1, which is achieved by lncRNA GAS5 that sponges miR-665. Our findings may provide a new treatment strategy for aging-related cardiovascular diseases.
登录
查看更多内容
影响因子：
3.7
作者：
Chaterji S;Lam CH;Ho DS;Proske DC;Baker AB
通讯作者：
Baker AB
影响因子：
2.9
作者：
Tan, Pan;Guo, Yong-Hong;Wang, Hai-Qin
通讯作者：
Wang, Hai-Qin
DOI：
10.1016/j.omtn.2016.11.005
发表时间：
2017-03-17
期刊：
Molecular therapy. Nucleic acids
影响因子：
--
作者：
Kim JH;Lee BR;Choi ES;Lee KM;Choi SK;Cho JH;Jeon WB;Kim E
通讯作者：
Kim E
影响因子：
4.5
作者：
Sathishkumar C;Prabu P;Mohan V;Balasubramanyam M
通讯作者：
Balasubramanyam M
影响因子：
1.5
作者：
Liu, Chuanzhen;Tang, Mengmeng;Cao, Guangqing
通讯作者：
Cao, Guangqing