Inhibitory role of ginsenoside Rb2 in endothelial senescence and inflammation mediated by microRNA-216a
Inhibitory role of ginsenoside Rb2 in endothelial senescence and inflammation mediated by microRNA-216a
复制标题
人参皂苷 Rb2 对 microRNA 介导的内皮衰老和炎症的抑制作用-216a
DOI：
10.3892/mmr.2021.12054
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发表时间：
2021-06-01
影响因子：
3.4
通讯作者：
Zhang, Weili
中科院分区：
文献类型：
--
作者：
Chen, Yutong;Wang, Shuting;Zhang, Weili
使用小化学分子靶向microRNA（miRs）已成为一种有前途的疾病治疗策略。miR-216 a通过Smad 3/NF-κ B信号通路被认为是内皮衰老和动脉粥样硬化的潜在治疗靶点。人参皂苷Rb 2（Ginsenoside Rb 2，Rb 2）是从植物人参（Panax ginseng）中提取的主要生物活性成分，是一种用途广泛的中药。在本研究中，通过基于其序列和结构特征的生物信息学分析，将Rb 2鉴定为与miR-216 a具有高评分。微量热泳实验进一步证明Rb 2对miR-216 a具有特异性结合亲和力，解离常数为17.6 μ M。在年轻和衰老的人脐静脉内皮细胞（HUVECs）以及人主动脉内皮细胞中，Rb 2降低了内源性miR-216 a的表达。接着，通过用pre-miR-216 a重组慢病毒（Lv-miR-216 a）感染建立HUVECs的复制性内皮衰老模型，并计算群体倍增水平（PDL）的数量。miR-216 a的稳定过表达诱导了早衰样表型，而衰老特征和衰老相关β-半乳糖苷酶（SA-β-gal）活性的增加在Rb 2处理后被逆转。与未经Rb 2处理的Lv-miR-216 a组相比，经Rb 2处理的Lv-miR-216 a转染的衰老PDL 25细胞中SA-β-gal阳性细胞的百分比降低了76%（P=0.01）。在机制上，miR-216 a抑制Smad 3蛋白表达，促进I κ B α降解，并激活NF-κ B应答基因，如血管细胞粘附分子1（VCAM 1），其促进内皮细胞向单核细胞的增殖。Rb 2处理显著抑制了miR-216 a的这些促炎作用。当Smad 3被小干扰RNA抑制时，由miR-216 a诱导的细胞间粘附分子1和VCAM 1的表达水平升高被显著逆转。总的来说，据我们所知，本研究首次证明Rb 2对内皮细胞衰老过程具有抗炎作用，并可能成为靶向miR-216 a的潜在治疗药物。
Targeting microRNAs (miRs) using small chemical molecules has become a promising strategy for disease treatment. miR-216a has been reported to be a potential therapeutic target in endothelial senescence and atherosclerosis via the Smad3/NF-kappa B signaling pathway. Ginsenoside Rb2 (Rb2) is the main bioactive component extracted from the plant Panax ginseng, and is a widely used traditional Chinese medicine. In the present study, Rb2 was identified to have a high score for miR-216a via bioinformatics analysis based on its sequence and structural features. The microscale thermophoresis experiment further demonstrated that Rb2 had a specific binding affinity for miR-216a and the dissociation constant was 17.6 mu M. In both young and senescent human umbilical vein endothelial cells (HUVECs), as well as human aortic endothelial cells, Rb2 decreased the expression of endogenous miR-216a. Next, a replicative endothelial senescence model of HUVECs was established by infection with pre-miR-216a recombinant lentiviruses (Lv-miR-216a) and the number of population-doubling level (PDL) was calculated. Stable overexpression of miR-216a induced a premature senescent-like phenotype, whereas the senescent features and increased activity of senescence-associated beta-galactosidase (SA-beta-gal) were reversed after Rb2 treatment. The percentage of SA-beta-gal-positive cells in senescent PDL25 cells transfected with Lv-miR-216a was decreased 76% by Rb2 treatment compared with the Lv-miR-216a group without Rb2 treatment (P=0.01). Mechanistically, miR-216a inhibited Smad3 protein expression, promoted I kappa B alpha degradation and activated NF-kappa B-responsive genes, such as vascular cell adhesion molecule 1 (VCAM1), which promoted the adhesiveness of endothelial cells to monocytes. These pro-inflammatory effects of miR-216a were significantly suppressed by Rb2 treatment. When Smad3 was suppressed by small interfering RNA, the elevated expression levels of intercellular adhesion molecule 1 and VCAM1 induced by miR-216a were significantly reversed. Collectively, to the best of our knowledge, the present study demonstrated for the first time that Rb2 exerted an anti-inflammation effect on the process of endothelial cell senescence and could be a potential therapeutic drug by targeting miR-216a.