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目的探讨雌激素受体对血管平滑肌细胞(VSMCS)早衰的影响及其机制。方法体外培养的第2～3代雌性SD大鼠VSMCS分为实验组和对照组,在有或无不同浓度(10-10mol/L～10-8 mol/L)雌激素受体激动剂(PPT/DPN)存在时,用150μmol/L H2O2诱导早衰。采用流式细胞术检测衰老相关标志物(DcR2)、Pull down as-say,Western blotting等方法检测原癌基因Ras的表达或活性。结果流式细胞术检测显示,H2O2组血管平滑肌细胞DcR2表达率显著增高(P﹤0.05)。H2O2+PPT及H2O2+DPN各浓度组血管平滑肌细胞DcR2表达率均低于H2O2组,且随激动剂浓度增高而降低,但不同激动剂之间无显著差别(P﹥0.05)。H2O2组细胞内GTP-Ras/Total Ras比值显著高于对照组(P﹤0.05),在有10-8 mol/L PPT或DPN存在时,细胞内的GTP-Ras/Total Ras比值明显降低,但仍高于对照组。H2O2+PPT组和H2O2+DPN组之间差异无统计学意义(P﹥0.05)。结论在介导雌激素的抗早衰效应上,雌激素受体a(ERa)和雌激素受体b(ERb)可能具有同等效应;雌激素抗早衰效应机制之一可能是抑制原癌基因Ras的激活。
Objective To investigate the effect of estrogen receptor on premature aging of vascular smooth muscle cells (VSMCS) and its mechanism. Methods VSMCs from the second to third passage of female SD rats cultured in vitro were divided into experimental group and control group, with or without different concentration (10-10mol / L ~ 10-8mol / L) of estrogen receptor agonist / DPN), induced premature aging with 150μmol / L H2O2. Apoptosis-related markers (DcR2), Pull down as-say and Western blotting were used to detect the expression or activity of Ras. Results Flow cytometry showed that the expression of DcR2 in H2O2 group was significantly increased (P <0.05). The expression of DcR2 in H2O2 + PPT and H2O2 + DPN groups was lower than that in H2O2 group, and decreased with increasing concentration of agonist, but there was no significant difference among different agonists (P> 0.05). The ratio of GTP-Ras / Total Ras in H2O2 group was significantly higher than that in control group (P <0.05). The ratio of GTP-Ras / Total Ras in cells treated with 10-8 mol / L PPT or DPN was significantly decreased Still higher than the control group. There was no significant difference between H2O2 + PPT group and H2O2 + DPN group (P> 0.05). Conclusions Estrogen receptor a (ERa) and estrogen receptor b (ERb) may have the same effect in mediating the anti-premature decay of estrogen. One of the mechanisms of estrogen anti-premature decay may be the inhibition of protooncogene Ras activation.