Division of Nephrology, Pudong Medical Center, Shanghai Pudong Hospital, Fudan University, 2800 Gong Wei Road, Shanghai, China.
Hemodialysis Center, Bao Shan Branch of No. 1, People's Hospital, Shanghai Jiao Tong University, Shanghai, China.
Mol Cell Biochem. 2018 Feb;439(1-2):131-140. doi: 10.1007/s11010-017-3142-6. Epub 2017 Sep 4.
Epigallocatechin gallate (EGCG), a bioactive ingredient of green tea, plays a protective role in the cardiovascular system. Homocysteine (Hcy) is a major risk factor for chronic kidney disease and cardiovascular disease. The present study aimed to investigate the role of EGCG in Hcy-induced proliferation of vascular smooth muscle cells (VSMCs) and its underlying mechanism. We also explored the roles of rennin-angiotensin system (RAS), extracellular signal-regulated kinases (ERK1/2), and p38 mitogen-activated protein kinase (p38 MAPK) in this process. Human aortic smooth muscle cells (HASMCs) were treated with different drugs for different periods. The proliferation rate of HASMCs was detected using the CCK-8 and BrdU labeling assays. The Western blot assay was used to determine the expression levels of angiotensin II type 1 receptor (AT-1R), ERK1/2, and p38 MAPK. Compared with the control group, the HASMCs treated with Hcy at different doses (100, 200, 500, and 1000 µM) showed significantly increased proliferation. Hcy increased the expression of AT-1R, whereas EGCG decreased the protein expression of AT-1R. Furthermore, we found that Hcy-induced expression of p-ERK1/2 and p-p38MAPK was dependent on AT-1R. Compared with Hcy (500 µM)-treated cells, EGCG (20 µM)-treated cells showed decreased proliferation as well as expression of AT-1R, p-ERK1/2, and p-p38MAPK. In addition, HASMC proliferation was suppressed by the addition of an AT-1R blocker (olmesartan), an ERK1/2 inhibitor (PD98059), and a p38MAPK inhibitor (SB202190). EGCG can inhibit AT-1R and affect ERK1/2 and p38MAPK signaling pathways, resulting in the decrease of VSMC proliferation induced by Hcy.
没食子酸表没食子儿茶素酯(EGCG)是绿茶中的一种生物活性成分,在心血管系统中发挥保护作用。同型半胱氨酸(Hcy)是慢性肾脏病和心血管疾病的主要危险因素。本研究旨在探讨 EGCG 在 Hcy 诱导的血管平滑肌细胞(VSMCs)增殖中的作用及其潜在机制。我们还探讨了肾素-血管紧张素系统(RAS)、细胞外信号调节激酶(ERK1/2)和 p38 丝裂原活化蛋白激酶(p38 MAPK)在这一过程中的作用。用不同的药物处理不同时期的人主动脉平滑肌细胞(HASMCs)。用 CCK-8 和 BrdU 标记检测 HASMCs 的增殖率。用 Western blot 检测血管紧张素 II 型 1 受体(AT-1R)、ERK1/2 和 p38 MAPK 的表达水平。与对照组相比,用不同剂量(100、200、500 和 1000µM)Hcy 处理的 HASMCs 增殖明显增加。Hcy 增加了 AT-1R 的表达,而 EGCG 降低了 AT-1R 的蛋白表达。此外,我们发现 Hcy 诱导的 p-ERK1/2 和 p-p38MAPK 表达依赖于 AT-1R。与 Hcy(500µM)处理的细胞相比,EGCG(20µM)处理的细胞增殖减少,AT-1R、p-ERK1/2 和 p-p38MAPK 的表达也减少。此外,用 AT-1R 阻滞剂(奥美沙坦)、ERK1/2 抑制剂(PD98059)和 p38MAPK 抑制剂(SB202190)抑制 HASMC 增殖。EGCG 可抑制 AT-1R,影响 ERK1/2 和 p38MAPK 信号通路,从而减少 Hcy 诱导的 VSMC 增殖。