脂多糖对大鼠星形胶质细胞Ski蛋白表达的影响

doi:10.3969/j.issn.1006-9771.2017.05.005

《中国康复理论与实践》 ›› 2017, Vol. 23 ›› Issue (5): 514-519.doi: 10.3969/j.issn.1006-9771.2017.05.005

脂多糖对大鼠星形胶质细胞Ski蛋白表达的影响

赵鑫^1,2, 郭永强^1,2, 王明^1,2, 陈铁戈^1,2, 伍亚民³, 张海鸿¹

1.兰州大学第二医院骨科,甘肃兰州市 730030;
2.甘肃省骨关节疾病研究重点实验室,甘肃兰州市 730030;
3.第三军医大学大坪医院野战外科研究所三室,创伤、烧伤与复合伤国家重点实验室,重庆市 400042。

收稿日期:2017-02-13 修回日期:2017-03-13 出版日期:2017-05-25 发布日期:2017-05-24
通讯作者: 张海鸿,主任医师,副教授,硕士研究生导师,主要研究方向：脊柱外科。E-mail: zhanghaihong1968@sina.com。
作者简介:赵鑫(1991-),男,汉族,山东潍坊市人,硕士研究生,主要研究方向：脊柱脊髓损伤。
基金资助:
国家自然科学基金项目(No.30772299)

Effect of Lipopolysaccharide on Ski Expression in Rats' Astrocytes

ZHAO Xin^1,2, GUO Yong-qiang^1,2, WANG Ming^1,2, CHEN Tie-ge^1,2, WU Ya-min³, ZHANG Hai-hong¹

1. Department of Orthopedics, the Second Clinical Medical College of Lanzhou University, Lanzhou, Gansu 730030, China;
2. Key Laboratory of Orthopedics of Gansu Province, Lanzhou, Gansu 730030, China;
3. State Key Laboratory of Trauma, Burns and Combined Injury, the Third Department of Research Institute of Surgery, Daping Hospital, Third Military University, Chongqing 400042, China

Received:2017-02-13 Revised:2017-03-13 Published:2017-05-25 Online:2017-05-24
Contact: ZHANG Hai-hong. E-mail: zhanghaihong1968@sina.com

摘要/Abstract

摘要： 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。

关键词: ski蛋白, 脂多糖, 炎症, 星形胶质细胞, 大鼠

Abstract: 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.

Key words: ski, lipopolysaccharide, inflammation, astrocyte, rats

中图分类号:

R364.5

赵鑫, 郭永强, 王明, 陈铁戈, 伍亚民, 张海鸿. 脂多糖对大鼠星形胶质细胞Ski蛋白表达的影响[J]. 《中国康复理论与实践》, 2017, 23(5): 514-519.

ZHAO Xin, GUO Yong-qiang, WANG Ming, CHEN Tie-ge, WU Ya-min, ZHANG Hai-hong. Effect of Lipopolysaccharide on Ski Expression in Rats' Astrocytes[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2017, 23(5): 514-519.

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脂多糖对大鼠星形胶质细胞Ski蛋白表达的影响

Effect of Lipopolysaccharide on Ski Expression in Rats' Astrocytes

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