olomoucine对星形胶质细胞增殖的影响及机制研究

目的:观察细胞周期抑制剂olomoucine对培养星形胶质细胞(AS)机械损伤后增殖的影响,并探讨其作用机制。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),随机分为对照组、划痕组和干预组,划痕组和干预组均继续培养,干预组给予olomoucine干预。利用免疫荧光细胞化学方法观察损伤后BrdU表达;利用Westernblot印迹分析损伤后PCNA、p27的表达。结果:划痕损伤刺激12h开始出现损伤边缘区AS胞体肥大、数目增加,BrdU阳性细胞比率和PCNA表达水平较对照组增高(P<0.05),p27表达水平较对照组下降(P<0.05);干预组的BrdU阳性细胞比率和PCNA表达水平较划痕组明显减少(P<0.05),p27表达上调(P<0.05)。结论:损伤刺激可促使AS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抑制其反应性增生。

The Effect of Olomoucine on Cultured Astrocyte Proliferation and its Mechanism Research

Objective:To observe the effect of CDK inhibitor-olmoucine on cultured astrocyte proliferation after mechanical injury and to explore the mechanism.Methods:The model of mechanical injury on cultured purified astrocytes was established.The cultured cells were randomly divided into 3 equal groups:control group,scratch group(scratch vehicle),and olomoucine group(scratch olomoucine).Immunofluorescence(IF)staining was used to detect the expression of glial fibriliary acidic protein(GFAP)and BrdU.Western bolt was used to detect the expression of PCNA and p27.Results:The "scratch-wound injury" can induce AS proliferation.From 12 hours after injury,AS around the injury area appeared hypertrophic and proliferative.The percent of the BrdU-positive cells and the expression of PCNA increased(P<0.05),while the expression of p27 decreased(P<0.05).In the olomoucine group,the percent of the BrdU-positive cells and the expression of PCNA were be inhibited significantly by olomoucine(P<0.05).Conclusion:As a response of injury,AS entered cell cycle and appeared reactive gliosis.The cell cycle inhibitor,olomoucine can inhibit injury-induced reactive gliosis by regulating cell cycle.