Olomoucine对星形胶质细胞增殖及活化分泌的影响

EFFECT OF CDK INHIBITOR OLOMOUCINE ON ASTROCYTE ACTIVATION AND PROLIFERATION

目的观察细胞周期抑制剂olomoucine对培养星形胶质细胞机械损伤后增殖和活化分泌的影响。方法建立体外培养大鼠纯化的AS物理损伤模型(划痕损伤模型),分为对照组、划痕组和olomoucine干预组。利用免疫荧光细胞化学方法观察损伤后GFAP表达;利用RT-PCR观察细胞因子IL-6和TNF-a的表达情况。结果划痕损伤刺激能使培养AS反应性增生活化。损伤早期出现IL-6、TNF-α表达水平增高,从损伤后12h开始出现损伤边缘区AS胞体肥大,数目增加;给予olomoucine干预后,细胞数目减少、体积明显减小,损伤后IL-6和TNF-α表达也显著下降。结论损伤刺激可促使AS活化,并发生反应性胶质增生;CDK选择性细胞周期抑制剂olomoucine可以通过调控细胞周期,有效抑制损伤后星形胶质细胞的反应性增生,并能对AS的活化起到抑制作用。

Objective To determine the effect of olomoucine inhibitor of cyclin-dependent protein kinases on the activation and proliferation of cultured astrocyte after mechanical injury. Methods The model of mechanical injury on cultured purified astrocytes was established. The cultured cells were randomly divided into 3 groups： control group, scratch group （scratch ＋ vehicle）, and olomoucine group （scratch ＋ olomoucine）. Immunofluorescence （IF） staining was used to detect the expression of glial fibrillary acidic protein （GFAP）. RT-PCR was used to detect the mRNA level of IL-6 and TNF-α. Results The ＂ scratch-wound injury＂ induced AS proliferation and activation. In the early phase of scratch injury, the mRNA level of IL-6 and TNF-α increased. From 12 h after injury, AS around the injury area showed hypertrophy and proliferation. In the olomoucine group, the number of GFAP positive cells decreased significantly. Moreover, olomoucine inhibited the expression of IL-6 and TNF-α. Conclusion As a response to injury, AS enter the cell cycle and showed reactive gliosis. The cell cycle inhibitor olomoucine can inhibit injury-induced reactive gliosis and astrocyte activation by regulating the cell cycle.