School of Medicine, Nankai University, Tianjin, China; Department of Cardiology, The Sixth Medical Center, Chinese PLA General Hospital, Beijing, 100853, China.
Department of Cardiology, The Sixth Medical Center, Chinese PLA General Hospital, Beijing, 100853, China.
Biochem Biophys Res Commun. 2024 Feb 12;696:149391. doi: 10.1016/j.bbrc.2023.149391. Epub 2023 Dec 28.
According to former research, the atherosclerotic plaque is thought to be aggravated by intraplaque neovessels (IPN) and intraplaque hemorrhage (IPH). Intriguingly, a lower incidence of IPH was found in plaque treated with melatonin. In this study, we attempted to investigate the impact and underlying mechanism regarding the influences of melatonin upon IPN.
A mouse model was established by subjecting the high fat diet (HFD)-fed ApoE-/- mice to tandem stenosis (TS) surgery with melatonin and GW9662, a PPARγ antagonist, being given by gavage. In vitro experiment was conducted with HUVECs exposing to according treatments of VEGF, melatonin, GW9662, or Y27632.
Plaque and IPN were attenuated by treatment with melatonin, which was then reversed by blocking PPARγ. Western blotting results showed that melatonin increased PPARγ and decreased RhoA/ROCK signaling in carotid artery. Elevated RhoA/ROCK signaling was observed in melatonin-treated mice when PPARγ was blocked. In accordance with it, experiments using protein and mRNA from HUVECs revealed that melatonin inhibited the RhoA/ROCK signaling by enhancing PPARγ. According to in vitro study, melatonin was able to inhibit cell migration and angiogenesis, which was aborted by GW9662. Blockage of ROCK using Y27632 was able to cease the effect of GW9662 and restored the suppression on cell migration and angiogenesis by melatonin.
Our study demonstrates that melatonin is able to curb development of plaque and IPN formation by inhibiting the migration of endothelial cells via PPARγ- RhoA-ROCK pathway. That provides a therapeutic potential for both melatonin and PPARγ agonist targeting IPN, IPH, and atherosclerotic plaque.
根据之前的研究,斑块内新生血管(IPN)和斑块内出血(IPH)被认为会加重动脉粥样硬化斑块。有趣的是,褪黑素治疗的斑块中 IPH 的发生率较低。在这项研究中,我们试图研究褪黑素对 IPN 的影响及其潜在机制。
通过对高脂饮食(HFD)喂养的 ApoE-/- 小鼠进行串联狭窄(TS)手术,建立了一个小鼠模型,并用灌胃给予褪黑素和 PPARγ 拮抗剂 GW9662。在体外实验中,用 HUVECs 暴露于 VEGF、褪黑素、GW9662 或 Y27632 等相应处理。
褪黑素治疗可减轻斑块和 IPN,阻断 PPARγ 则可逆转这一作用。Western blot 结果显示,褪黑素增加了颈动脉中的 PPARγ,降低了 RhoA/ROCK 信号。当阻断 PPARγ 时,在褪黑素处理的小鼠中观察到 RhoA/ROCK 信号升高。与之相符的是,使用 HUVECs 的蛋白质和 mRNA 的实验表明,褪黑素通过增强 PPARγ 抑制了 RhoA/ROCK 信号。根据体外研究,褪黑素能够抑制细胞迁移和血管生成,而 GW9662 则可以阻止这一作用。使用 Y27632 阻断 ROCK 可以停止 GW9662 的作用,并恢复褪黑素对细胞迁移和血管生成的抑制作用。
我们的研究表明,褪黑素通过抑制内皮细胞的迁移,通过 PPARγ-RhoA-ROCK 通路抑制斑块和 IPN 的形成。这为褪黑素和 PPARγ 激动剂针对 IPN、IPH 和动脉粥样硬化斑块的治疗提供了潜力。