Ski在大鼠活化星形胶质细胞中表达的变化

Changes of Ski expression levels in rat activated astrocytes

目的:探究Ski在大鼠正常及活化星形胶质细胞中的表达及随时间变化的规律。方法:提取大鼠大脑皮质,分离星形胶质细胞,体外培养。采用LPS刺激和细胞划痕损伤2种方法活化星形胶质细胞,均采用Western blot法检测各个时点2种活化星形胶质细胞中Ski和胶质纤维酸性蛋白(GFAP)蛋白表达情况,利用间接免疫荧光法检测Ski蛋白在星形胶质细胞中的表达定位。结果:GFAP蛋白在星形胶质细胞中固有表达,LPS活化和划痕损伤后表达均上调。Ski在正常星形胶质细胞中呈低表达状态,1 mg/L LPS活化星形胶质细胞后,Ski表达开始增加,4 d时表达量最高(P<0.05),6 d时开始下降,但仍高于未活化组;细胞划痕损伤后Ski蛋白表达情况与上述情况高度一致。Ski在正常及LPS活化6 d星形胶质细胞中表达主要集中在胞核,LPS活化后2和4 d时在胞质中出现明确的Ski表达。结论:Ski蛋白表达于星形胶质细胞,并在活化星形胶质细胞中表达上调。由此,我们推测Ski蛋白可能调控星形胶质细胞的活化、增殖等过程。

AIM ： To explore the time-dependent change of Ski protein expression in normal and activated as-trocytes in rats. METHODS ： The astrocytes were obtained from rat cerebral cortex and cultured in vitro. The astrocytes were treated with LPS and scratch injury for activation. Western blot analysis was used to determine glial fibrillary acidic protein （GFAP） and Ski protein levels in activated astrocytes at a series of time points. The indirect immunofluorescence staining method was performed to detect the location of Ski protein in the astrocytes. RESULTS ： The protein of GFAP was naturally expressed in the astrocytes, beginning to increase after treated with LPS and scratch injury. Little protein expres-sion of Ski in the normal astrocytes was observed. The Ski protein expression began to increase after treated with 1 mg/L LPS,peaked at 4 d （ P 〈 0. 05 ） and then deceased, but was stills higher than that in the normal cells. The protein expres-sion level of Ski after scratch injury was highly consistent with above mentioned. Ski was mainly observed in the nucleus of the normal cells and the cells treated with LPS for 6 d , while it was observed in the cytoplasm 2 and 4 d after treated with LPS. CONCLUSION： The protein of Ski is expressed in the astrocytes, and the expression level is increased in activated astrocytes, mainly located in the nucelus. Ski may plays an essential roles in the processes of activation and proliferation of astrocytes.