Objective To explore the effect of ERK1/2 phosphorylation level on the proliferation and cell cycle of astrocytes in vitro. Methods Primary cultured neonatal SD rat astrocytes were divided into normal control group without any treatment and U0126 treatment group (U0126 group). Western blotting, click-iT Edu test and flow cytometry were performed to detect the phosphorylation level of ERK1/2, astrocgtes proliferation and the percentage of astrocytes in each cell cycle phase respectively. Results In the U0126 group ERK1/2 phosphorylation level (39. 13% ±6. 71%) was significantly lower than that of control group (100%). At 24 h, the proliferation cell percentage of astrocgtes in U0126 group [(2. 63±1. 14)%] was significantly lower than that of control group [(21. 43 ± 3. 81)%] 0 = 21.13, P<0. 01). In group control group and U0126 group, the proportion of astrocgtes in the G1 phase were (84.63 ± 1. 00) % and (93.67 ± 0.68)%, in the G2 phase were (2.08 ± 0.21)% and (3. 43 ± 0. 88)%, in the Sphase were (14. 21 ± 1. 14)% and (2. 90 ± 0. 23)%; all these results have statistically significant differences (t=12.91, P<0.01; t = 4. 35, P<0.05; t= 16. 87, P< 0. 01). Conclusions Reducing the level of ERK1/2 phosphorylation could inhibit the proliferation of astrocytes, they were arrested at G1 phase by inhibiting the cell cycle transformation from G1 phase to S phase and G2 phase by inhibiting the astrocytes division.%目的 探讨细胞外信号调节激酶(ERK1/2)的磷酸化水平对体外星形胶质细胞(Ast)增殖及其细胞周期的影响.方法 将培养成熟的大鼠原代Ast分为两组,其中一组用ERK1/2磷酸化的特异性抑制剂U0126进行处理(U0126组),另一组不做处理,作为对照组.通过Western Blot技术、Click-iT Edu技术和流式细胞术分别检测两组Ast中ERK1/2磷酸化水平、Ast增殖细胞百分比及各期细胞百分比.结果 U0126组ERK1/2的磷酸化水平(39.13％±6.71％)明显低于对照组(100％).U0126处理24 h后,U0126组Ast增殖细胞百分比[(2.63±1.14)％]低于对照组[(21.43±3.81)％](t=21.13,P＜0.01).G1期U0126组Ast[(93.67±0.68)％]高于对照组[(84.63±1.00)％](t=12.91,P＜0.01)；S期U0126组Ast[(2.90±0.23)％]低于对照组[(14.21±1.14)％] (t=16.87,P＜0.01);G2期U0126组Ast[(3.43±0.88)％]高于对照组[(2.08±0.21％)％](t=4.35,P＜0.05).结论 降低ERK1/2的磷酸化水平可抑制Ast的增殖,并将其阻断在G1期,抑制Ast从G1期向S期的转化,同时抑制其分裂将其阻断在G2期.
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