G蛋白偶联雌激素受体介导17β-雌二醇抗血管平滑肌细胞氧化应激性衰老

17β-estradiol protects vascular smooth muscle cells from oxidative stress-induced senescence via G protein-coupled estrogen receptor

目的探讨G蛋白偶联雌激素受体(GPER)能否介导雌激素抗大鼠血管平滑肌细胞(vascular smooth muscle cells,VSMCs)氧化应激性衰老效应。方法取体外培养第3~4代VSMCs,以150μmol/L H_2O_2处理2h建立细胞衰老模型。用10^(-8)mol/L17β-雌二醇(E_2)、10^(-6)mol/LGPER激动剂G1和抑制剂G15分别处理VSMCs。衰老相关β-半乳糖苷酶(SA-β-Gal)染色法检测各组细胞SA-β-Gal染色阳性率;流式细胞术检测各组细胞周期变化;RT-qPCR和Western blot检测GPERmRNA和蛋白表达变化。结果 H_2O_2使VSMCs SA-β-Gal染色阳性率明显增加,细胞周期停滞于G0/G1期,GPER mRNA和蛋白水平降低,E2和G1对H_2O_2的这些效应具有抑制作用,而G15可阻断E2和G1对H_2O_2上述效应的抑制作用。结论雌激素依赖GPER发挥抗血管平滑肌细胞氧化应激性衰老效应。

Objective To explore whether estrogen protects vascular smooth muscle cells （VSMCs） from oxidative stress-in- duced senescence via G protein-coupled estrogen receptor （GPER）. Methods The 3 - 4 passages of VSMCs cultured in vitro were pre- treated with 150μM H202 for 2 hours to establish the senescence model. They were then divided into different groups to be treated with 1010^-8mol/L 17β-estradiol（E2）, 10^-6M GPER agonist G1, 10^-6M GPER antagonist G15, G15 ＋ E2, G15 ＋ G1, respectively. The senes- cence of VSMCs in each group was detected by senescence-associated beta-galactosidase （ SA-13-Gal） staining, the change in cell cycle by flow cytometry, and the expression of GPER mRNA and protein by RT-qPCR and western blot. Results H202 treatment significantly increased the percentage of SA-β-Gal-positive VSMCs, the cell cycle was arrested in G0/G1 phase, and the expression of GPER mRNA and protein decreased greatly. E2 and G1 inhibited these effects of n202 on VSMCs, while G15 could block the inhibitory effect of E2 and G1. Conclusion Estrogen protects vascular smooth muscle cells from oxidative stress-induced senescence via GPER.