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为了研究血管紧张素 (Ang )的抗内皮细胞增生作用及其分子生物学调节机制 ,将健康胎儿脐静脉内皮细胞培养至第 3代 ,用 Ang 培养并在不同时间计数细胞密度。用 TUNEL 或 EL ISA方法证明 Ang 或 CGP42 112 A诱导内皮细胞凋亡。用 RT- PCR和 Western Blot检测 p2 1 / waf1m RNA和 P2 1 蛋白的表达并测定条带的光密度。结果表明 ,在 Ang 培养后不同时间 ,细胞密度比对照减少 30 %左右 (P<0 .0 1)。Ang 作用后可以诱导典型的凋亡 ,阳性率极显著高于阴性对照组并具有剂量依赖性。Ang 组或 CGP42 112 A组的 p2 1 / waf1m RNA表达分别是对照组的 (4 73.6 9± 39.97) %和 (391.5 8± 48.2 4) %(P<0 .0 1)。Ang 组的 P2 1 蛋白表达比对照组极显著地增加并具有时间依赖性。认为 Ang 具有抗内皮细胞增生作用 ,其细胞内机制是同时诱导凋亡和 p2 1 / waf1m RNA与 P2 1 蛋白的高表达 ,使细胞发生 G1 期阻滞 ,增殖受抑。
To investigate the anti-endothelial cell proliferation and molecular mechanisms of angiotensin (Ang) regulation, healthy human umbilical vein endothelial cells were cultured to passage 3, cultured in Ang and cell density was counted at different times. Angio or CGP42 112 A-induced endothelial cell apoptosis was demonstrated using TUNEL or EL ISA methods. The expression of p2 1 / waf1m RNA and P2 1 protein was detected by RT-PCR and Western Blot and the optical density of the bands was determined. The results showed that at different time points after Ang culture, the cell density was reduced by 30% (P <0.01) compared with the control. Apoptosis induced by Ang can induce typical apoptosis, the positive rate was significantly higher than the negative control group and dose-dependent. Expression of p2 1 / waf1m RNA in Ang group or CGP42 112 A group was (4 73.6 9 ± 39.97)% and (391.5 8 ± 48.2 4)% (P <0.01) in the control group, respectively. The P2 1 protein expression in Ang group was significantly increased and time-dependent compared with the control group. It is considered that Ang has an anti-endothelial cell proliferation effect. The intracellular mechanism induces apoptosis and the overexpression of p2 1 / waf1m RNA and P2 1 protein at the same time, which results in G1 arrest and proliferation inhibition.