Department of Pharmacology, College of Medicine, Yeungnam University, 170 Hyunchung-Ro, Nam-Gu, Daegu 42415, Korea.
Department of Microbiology, Yeungnam University, 170 Hyunchung-Ro, Nam-Gu, Daegu 42415, Korea.
Molecules. 2021 Jan 26;26(3):640. doi: 10.3390/molecules26030640.
Vascular smooth muscle cell (VSMC) phenotype switching from contractile to synthetic is essential for proliferation and migration in vascular pathophysiology. Connective tissue growth factor (CTGF) is a matricellular protein involved in cell adhesion, migration, and proliferation. Kahweol, a diterpene molecule in arabica coffee beans, has been reported to have anti-inflammatory, antiproliferative, and apoptotic effects in many cells. However, in VSMCs, the effects of kahweol on CTGF activities have not been investigated. Thus, in this study, the effects and associated mechanisms of kahweol in CTGF-dependent phenotype switching and migration in VSMCs were examined. Experiments were performed on primary rat aortic smooth muscle cells and a rat VSMC line, A7r5. Western blot analysis was used to determine the protein levels. The mRNA levels of synthetic markers were measured by qRT-PCR. Migration of VSMCs was evaluated by wound healing and transwell assays. Kahweol reduced the angiotensin II (Ang II)-induced CTGF expression. Further, kahweol inhibited expressions of synthetic phenotype markers of VSMC. The kahweol-reduced synthetic marker protein levels were reversed by the administration of rCTGF. However, expressions of contractile phenotype markers of VSMC were not affected. Kahweol suppressed Ang II-stimulated VSMC migration. Moreover, kahweol downregulated Ang II-induced p-FAK, p-Erk, and Yes-associated protein (YAP) protein expressions. Taken together, in Ang II-stimulated VSMCs, kahweol inhibited CTGF-dependent synthetic phenotype switching and migration, with focal adhesion kinase (FAK), Erk, and YAP involved in the underlying mechanisms of the kahweol effects. These results suggest that kahweol has a potential as a therapeutic agent to inhibit CTGF, which is a molecular target in sclerogenic vascular disease.
血管平滑肌细胞(VSMC)从收缩型向合成型表型的转换对于血管病理生理学中的增殖和迁移是必不可少的。结缔组织生长因子(CTGF)是一种细胞外基质蛋白,参与细胞黏附、迁移和增殖。阿拉伯咖啡豆中的二萜分子卡瓦醇已被报道在许多细胞中具有抗炎、抗增殖和促凋亡作用。然而,在 VSMCs 中,卡瓦醇对 CTGF 活性的影响尚未得到研究。因此,在这项研究中,研究了卡瓦醇在 CTGF 依赖性表型转换和 VSMCs 迁移中的作用及其相关机制。实验在原代大鼠主动脉平滑肌细胞和大鼠 VSMC 系 A7r5 上进行。通过 Western blot 分析测定蛋白水平。通过 qRT-PCR 测定合成标记物的 mRNA 水平。通过划痕愈合和 Transwell 测定评估 VSMC 的迁移。卡瓦醇降低了血管紧张素 II(Ang II)诱导的 CTGF 表达。此外,卡瓦醇抑制了 VSMC 合成表型标志物的表达。用 rCTGF 处理可逆转卡瓦醇降低的合成标志物蛋白水平。然而,VSMC 的收缩表型标志物的表达不受影响。卡瓦醇抑制了 Ang II 刺激的 VSMC 迁移。此外,卡瓦醇下调了 Ang II 诱导的 p-FAK、p-Erk 和 Yes 相关蛋白(YAP)蛋白表达。总之,在 Ang II 刺激的 VSMCs 中,卡瓦醇抑制了 CTGF 依赖性合成型表型转换和迁移,其中粘着斑激酶(FAK)、Erk 和 YAP 参与了卡瓦醇作用的潜在机制。这些结果表明,卡瓦醇具有作为治疗剂抑制 CTGF 的潜力,CTGF 是硬皮病血管疾病的一个分子靶标。