Zhang Manli, Zhang Manna, Wang Wenli, Chen Hui, Wang Xia, Zhao Kun, Li Ze, Xu Jiangqing, Tong Fei
Department of Critical Care Medicine, The Second Hospital of Hebei Medical University Shijiazhuang, Hebei, China.
Department of Clinical Laboratory, The Second Hospital of Hebei Medical University Shijiazhuang, Hebei, China.
Am J Transl Res. 2022 Sep 15;14(9):6123-6136. eCollection 2022.
In vascular remodeling diseases, proliferation and inflammation of vascular smooth muscle cells (VSMCs) constitute the basic pathologic processes. Dehydroepiandrosterone (DHEA) exerts a protective effect on the cardiovascular system, but the molecular mechanism is unclear.
The plasma DHEA was measured using enzyme-linked immunosorbent assay (ELISA) kits. The neointima hyperplasia was assessed by hematoxylin/eosin staining. MiRNA microarray analysis was used to compare the influence of Ang II and DHEA on miRNA expression profiles in VSMCs. Cell counting and MTS assay were used to evaluate the effect of Ang II, DHEA and miR-486a-3p on VSMCs proliferation. qRT-PCR was performed to detect the expression of miR-486a-3p, PCNA, IL-1β and NLRP3. Western blot analysis was performed to detect the expressions of PCNA, IL-1β and NLRP3 after miR-486a-3p was knocked down or overexpressed in VSMCs.
DHEA suppressed neointimal and VSMCs proliferation and inflammation. Using miRNA microarray analysis, we found that DHEA upregulated the expression of miR-486a-3p in VSMCs. Further experiments indicated that DHEA promoted miR-486a-3p expression in VSMCs and in the vascular intima. Gain- and loss-of-function experiments revealed that transfection of miR-486a-3p mimic inhibited proliferation and inflammation of VSMCs, which improved intimal hyperplasia. On the contrary, deletion of miR-486a-3p promoted VSMCs proliferation and inflammation. Furthermore, DHEA suppressed NOD-like receptor family pyrin domain containing 3 (NLRP3) expression and reduced VSMCs proliferation and inflammation. Importantly, DHEA inhibited NLRP3 expression via miR-486a-3p in VSMCs.
DHEA inhibited VSMCs and vascular intimal proliferation and inflammation by regulating the miR-486a-3p/NLRP3 axis. Therefore, DHEA might be a candidate cardiovascular protective agent in the future.
在血管重塑疾病中，血管平滑肌细胞（VSMC）的增殖和炎症构成了基本的病理过程。脱氢表雄酮（DHEA）对心血管系统具有保护作用，但其分子机制尚不清楚。
使用酶联免疫吸附测定（ELISA）试剂盒测量血浆DHEA。通过苏木精/伊红染色评估内膜增生。采用miRNA微阵列分析比较血管紧张素II（Ang II）和DHEA对VSMC中miRNA表达谱的影响。使用细胞计数和MTS测定法评估Ang II、DHEA和miR-486a-3p对VSMC增殖的影响。进行qRT-PCR检测miR-486a-3p、增殖细胞核抗原（PCNA）、白细胞介素-1β（IL-1β）和NOD样受体家族含pyrin结构域蛋白3（NLRP3）的表达。在VSMC中敲低或过表达miR-486a-3p后，进行蛋白质免疫印迹分析检测PCNA、IL-1β和NLRP3的表达。
DHEA抑制内膜和VSMC的增殖及炎症。通过miRNA微阵列分析，我们发现DHEA上调了VSMC中miR-486a-3p的表达。进一步实验表明，DHEA促进VSMC和血管内膜中miR-486a-3p的表达。功能获得和功能缺失实验表明，转染miR-486a-3p模拟物可抑制VSMC的增殖和炎症，改善内膜增生。相反，缺失miR-486a-3p则促进VSMC的增殖和炎症。此外，DHEA抑制NLRP3的表达，减少VSMC的增殖和炎症。重要的是，DHEA在VSMC中通过miR-486a-3p抑制NLRP3的表达。
DHEA通过调节miR-486a-3p/NLRP3轴抑制VSMC和血管内膜的增殖及炎症。因此，DHEA未来可能是一种心血管保护剂候选药物。
