目的 探讨内毒素脂多糖对大鼠星形胶质细胞ski蛋白表达规律及胞定位的影响。方法 新生3 d内Sprague-Dawley大鼠,取大脑皮质分离星形胶质细胞,体外培养。采用0 μg/ml、0.001 μg/ml、0.01 μg/ml、0.1 μg/ml、1 μg/ml、10 μg/ml、100 μg/ml 脂多糖诱导6 h;采用0.1 μg/ml脂多糖诱导0 d、2 d、4 d、6 d、8 d。Western blotting法检测星形胶质细胞中ski蛋白的表达,间接免疫荧光双标法检测ski在星形胶质细胞中的定位。结果 0.1 μg/ml脂多糖诱导下,ski蛋白表达最高(F=46.656, P<0.001)。0.1 μg/ml脂多糖诱导4 d时最高,6 d时逐渐降低。在无脂多糖诱导及脂多糖诱导6 d时,ski蛋白主要表达于细胞核;脂多糖诱导2 d、4 d时,细胞质中出现ski表达。结论 脂多糖可诱导星形胶质细胞ski蛋白表达,可能参与炎症反应。
Objective To explore how and where ski expresses under lipopolysaccharide (LPS) in rats' astrocytes. Methods Astrocytes were obtained from cerebral cortex of a newborn (within 3 days) Sprague-Dawley rat and cultured in vitro. Astrocytes were cultured with LPS in concentration of 0 μg/ml, 0.001 μg/ml, 0.01 μg/ml, 0.1 μg/ml, 1 μg/ml, 10 μg/ml and 100 μg/ml for six hours; and cultured with LPS in concentration of 0.1 μg/ml for 0 day, 2 days, 4 days, 6 days and 8 days. The level of ski was determined with Western blotting, and the location of ski was detected with indirect immunofluorescent staining. Results The expression of ski was induced by LPS, especially in the concentration of 0.1 μg/ml. The expression of ski induced with 0.1 μg/ml LPS peaked at 4 days of inducement and then decreased. Ski was mainly observed in nuclear in the normal astrocytes and the astrocytes induced with 0.1 μg/ml LPS for 6 days. However, it was observed in cytoplasm 2 and 4 days of inducement. Conclusion LPS could induce the expression of ski in rats' astrocytes, which may participate in inflammation.
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