Department of Clinical Medicine, Jishou University School of Medicine, Jishou, Hunan 416000, P.R. China.
Department of Nursing, Jishou University School of Medicine, Jishou, Hunan 416000, P.R. China.
Mol Med Rep. 2021 Jul;24(1). doi: 10.3892/mmr.2021.12123. Epub 2021 Apr 28.
The phenotypes and mechanisms underlying the proliferation and migration of vascular smooth muscle cells (VSMCs) induced by oleic acid (OA) are not completely understood. Therefore, the aim of the present study was to further elucidate the effects of OA on the proliferation and migration of VSMCs. Using A7r5 cells, the hepatocyte growth factor (HGF) inhibitor PHA665752 and the p38 MAPK inhibitor SB203580 were utilized, and Cell Counting Kit‑8 (CCK‑8) assays, Transwell assays, flow cytometry, ELISAs, western blotting and reverse transcription‑quantitative PCR (RT‑qPCR) were conducted to assess the effects of OA. CCK‑8 assays indicated that OA promoted (at 5 and 50 mol/l) or inhibited (at 800 mol/l) A7r5 cell proliferation in a time‑ and concentration‑dependent manner (P<0.05). Transwell assays revealed that OA also promoted (at 50 mol/l) or inhibited (at 800 mol/l) A7r5 cell migration (P<0.05). Moreover, cell‑cycle analysis identified that 50 mol/l OA reduced the cellular population in the G/G phase and enhanced the cellular population in the S phase (P<0.05), whereas 800 mol/l OA increased the cell number in the G/G phase and decreased the cell number in the S phase (P<0.05). In addition, OA promoted (at 50 mol/l) or inhibited (at 800 mol/l) the expression level of HGF in A7r5 cells, as demonstrated via ELISA, western blotting and RT‑qPCR analyses (P<0.05). It was also found that OA promoted (at 50 mol/l) or inhibited (at 800 mol/l) the expression level of phosphorylated (p)‑p38 in A7r5 cells, as indicated by western blotting (P<0.05). Furthermore, the cell proliferation, migration and HGF expression induced by OA (50 mol/l) were mitigated by treatment with PHA665752 (0.1 mol/l) (P<0.05), and the cell proliferation, migration and p‑p38 expression induced by OA (50 mol/l) were mitigated by SB203580 (2 mol/l) (P<0.05). Thus, the results suggested that OA served a role in the proliferation and migration of VSMCs via HGF and the p38 MAPK pathway. Moreover, the proliferation and migration of VSMCs induced by OA was associated with increased expression levels of HGF and p‑p38. Taken together, OA, HGF and p38 MAPK may be potential therapeutic targets for the treatment of atherosclerosis.
油酸（OA）诱导的血管平滑肌细胞（VSMC）增殖和迁移的表型和机制尚不完全清楚。因此，本研究旨在进一步阐明 OA 对 VSMC 增殖和迁移的影响。使用 A7r5 细胞，利用肝细胞生长因子（HGF）抑制剂 PHA665752 和 p38 MAPK 抑制剂 SB203580，通过细胞计数试剂盒-8（CCK-8）检测、Transwell 检测、流式细胞术、ELISA、Western blot 和逆转录-定量 PCR（RT-qPCR）评估 OA 的作用。CCK-8 检测结果表明，OA 以时间和浓度依赖性方式促进（在 5 和 50 μmol/L 时）或抑制（在 800 μmol/L 时）A7r5 细胞增殖（P<0.05）。Transwell 检测结果表明，OA 还促进（在 50 μmol/L 时）或抑制（在 800 μmol/L 时）A7r5 细胞迁移（P<0.05）。此外，细胞周期分析表明，50 μmol/L OA 降低了 G/G 期细胞群体并增加了 S 期细胞群体（P<0.05），而 800 μmol/L OA 增加了 G/G 期细胞数量并减少了 S 期细胞数量（P<0.05）。此外，ELISA、Western blot 和 RT-qPCR 分析表明，OA 促进（在 50 μmol/L 时）或抑制（在 800 μmol/L 时）A7r5 细胞中 HGF 的表达水平（P<0.05）。还发现，OA 通过 Western blot（P<0.05）促进（在 50 μmol/L 时）或抑制（在 800 μmol/L 时）A7r5 细胞中磷酸化（p）-p38 的表达水平。此外，OA（50 μmol/L）诱导的细胞增殖、迁移和 HGF 表达通过 PHA665752（0.1 μmol/L）治疗得到缓解（P<0.05），OA（50 μmol/L）诱导的细胞增殖、迁移和 p-p38 表达通过 SB203580（2 μmol/L）治疗得到缓解（P<0.05）。因此，研究结果表明，OA 通过 HGF 和 p38 MAPK 通路在 VSMC 的增殖和迁移中发挥作用。此外，OA 诱导的 VSMC 增殖和迁移与 HGF 和 p-p38 的表达水平增加有关。综上所述，OA、HGF 和 p38 MAPK 可能是治疗动脉粥样硬化的潜在治疗靶点。
