小分子化合物 Y16 和 Rhosin 可以抑制自发性高血压大鼠血管平滑肌细胞中的钙敏化通路。
Small chemical compounds Y16 and Rhosin can inhibit calcium sensitization pathway in vascular smooth muscle cells of spontaneously hypertensive rats.
机构信息
Graduate Institute of Physiology, College of Medicine, National Taiwan University, Taipei, Taiwan.
Division of Cardiology, Department of Internal Medicine, National Taiwan University Hospital, Taipei, Taiwan.
出版信息
J Formos Med Assoc. 2021 Oct;120(10):1863-1868. doi: 10.1016/j.jfma.2021.03.031. Epub 2021 Apr 21.
BACKGROUND/PURPOSE: The small-molecule compounds Y16 and Rhosin can inhibit the activation of leukemia-associated Rho guanine nucleotide exchange factor (LARG) and small G-protein RhoA, respectively, in breast cancer cells and inhibit their growth and migration. However, it remains unclear whether they have inhibitory effects on the vascular smooth muscle cells (VSMCs) of spontaneously hypertensive rats (SHRs).
METHODS
Primary cultured VSMCs from SHRs were treated with different concentrations of Y16 or Y16 plus Rhosin for 24 h, followed by 10-min stimulation with 10 M angiotensin II (Ang II). The cells were then harvested, and the total protein was extracted. The co-immunoprecipitation method, Western blot analysis, and MTT assay were performed to determine the LARG-RhoA interaction, the protein levels of RhoA and MYPT1, and cell viability, respectively.
RESULTS
Y16 dose-dependently inhibited the LARG-RhoA complex formation induced by Ang II. With 50 μM of Y16, the effect of inhibition was statistically significant. Y16 also reduced the formation of phospho-MYPT1 stimulated by Ang II. With 5 μM of Y16, the inhibitory effect was statistically significant. When 25 μM of Y16 and 25 μM of Rhosin were combined, the inhibitory effect on LARG-RhoA interaction was statistically significant. When Y16 and Rhosin were combined, a significantly reduced concentration could effectively inhibit MYPT1 phosphorylation (2.5 μM compared with 5 μM for Y16 alone).
CONCLUSION
Treating SHR VSMCs with Y16 can suppress the activation of LARG, prevent LARG binding to RhoA, and decrease the phosphorylation of MYPT1, thus weakening the activation of the calcium (Ca) sensitization pathway in SHR VSMCs.
背景/目的：小分子化合物 Y16 和 Rhosin 分别可以抑制乳腺癌细胞中白血病相关 Rho 鸟嘌呤核苷酸交换因子（LARG）和小 G 蛋白 RhoA 的激活，并抑制其生长和迁移。然而，它们是否对自发性高血压大鼠（SHR）的血管平滑肌细胞（VSMCs）有抑制作用尚不清楚。
方法
用不同浓度的 Y16 或 Y16 加 Rhosin 处理原代培养的 SHR VSMCs 24 h，然后用 10 μM 血管紧张素 II（Ang II）刺激 10 min。收获细胞，提取总蛋白。采用共免疫沉淀法、Western blot 分析和 MTT 法分别检测 LARG-RhoA 相互作用、RhoA 和 MYPT1 蛋白水平以及细胞活力。
结果
Y16 呈剂量依赖性抑制 Ang II 诱导的 LARG-RhoA 复合物形成。用 50 μM 的 Y16 时，抑制作用具有统计学意义。Y16 还降低了 Ang II 刺激的磷酸化 MYPT1 的形成。用 5 μM 的 Y16 时，抑制作用具有统计学意义。当 25 μM 的 Y16 和 25 μM 的 Rhosin 联合使用时，对 LARG-RhoA 相互作用的抑制作用具有统计学意义。当 Y16 和 Rhosin 联合使用时，降低浓度即可有效抑制 MYPT1 磷酸化（Y16 单独使用时为 2.5 μM，而联合使用时为 5 μM）。
结论
用 Y16 处理 SHR VSMCs 可抑制 LARG 的激活，阻止 LARG 与 RhoA 结合，并降低 MYPT1 的磷酸化，从而减弱 SHR VSMCs 中钙（Ca）敏化途径的激活。
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