人参皂苷Rg1抑制脂多糖诱导BV-2小胶质细胞增殖

doi:10.16098/j.issn.0529-1356.2016.05.006

解剖学报 ›› 2016, Vol. 47 ›› Issue (5): 614-619.doi: 10.16098/j.issn.0529-1356.2016.05.006

人参皂苷Rg1抑制脂多糖诱导BV-2小胶质细胞增殖

姚玥伊^1,3艾青龙² 陈媛丽³ 尚群竺² 张汝江² 毕秀梅²孙俊^3*钟莲梅^2*

1. 昆明医科大学生物医学工程研究中心，昆明 650500;2. 昆明医科大学第一附属医院神经内科，昆明 650032;3. 昆明医科大学人体解剖学与组织学胚胎学系，昆明 650500

收稿日期:2016-02-01 修回日期:2016-05-20 出版日期:2016-10-06 发布日期:2016-10-06
通讯作者: 孙俊,钟莲梅 E-mail:sunjun6661@126.com
基金资助:
人参皂苷Rg1抑制小胶质细胞激活的信号转导机制研究;天麻素在癫痫持续状态中的保护作用及其信号机制研究;人参皂苷Rg1在癫痫持续状态中的保护作用及分子机制;Wnt/β-catenin信号通路在天麻素抑制小胶质细胞激活中的分子机制的研究

Effect of ginsenoside Rg1 on proliferation induced by lipopolysaccharide in BV-2 cells

YAO Yue-yi^1,3 AI Qing-long²CHEN Yuan-li³ SHANG Qun-zhu² ZHANG Ru-jiang²BI Xiu-mei² SUN Jun ^3* ZHONG Lian-mei ^2*

1. Kunming Medical University Biomedical Engineering Research Center，Kunming 650500，China; 2. Department of Neurology, the First Affiliated Hospital of Kunming Medical University，Kunming 650032，China; 3. Department of Anatomy, Histology and Embryology of Kunming Medical University，Kunming 650500，China

Received:2016-02-01 Revised:2016-05-20 Online:2016-10-06 Published:2016-10-06
Contact: SUN Jun,ZHONG Lian-mei E-mail:sunjun6661@126.com

摘要/Abstract

摘要：

目的探讨人参皂苷Rg1（Rg1）对脂多糖（LPS）诱导BV-2小胶质细胞中增殖细胞核抗原（PCNA）和细胞周期蛋白D1（cyclin D1）的调控作用及其对活化的BV-2小胶质细胞增殖的调控作用。方法体外培养BV-2小胶质细胞并分为空白对照组、LPS不同时间（1，3，6，8，12h）处理组及Rg1不同浓度（10、20和50μmol/L）预处理组。采用Western blotting检测细胞增殖相关因子PCNA蛋白和cyclin D1蛋白的表达变化；RT-PCR检测细胞增殖相关因子PCNA和cyclin D1 mRNA的表达变化；免疫荧光染色法观察细胞内cyclin D1蛋白表达变化。采用流式细胞术检测细胞周期的变化。结果 LPS诱导BV-2小胶质细胞PCNA和cyclin D1mRNA和蛋白表达上调，并呈时间依赖性；不同浓度Rg1预处理下调LPS诱导的BV-2小胶质细胞PCNA和cyclin D1的mRNA和蛋白表达。结论 Rg1通过下调LPS诱导的BV-2小胶质细胞PCNA和cyclin D1的表达水平来调控细胞周期进而抑制活化的小胶质细胞的增殖。

关键词: 人参皂苷Rg1, 脂多糖, 增殖, BV-2小胶质细胞, 增殖细胞核抗原, 细胞周期蛋白D1, 流式细胞术

Abstract:

Objective To investigate the possible role of ginsenoside Rg1 (Rg1) mediated in proliferating cell nuclear antigen (PCNA) and cyclin D1 of BV-2 cells stimulated by lipopolysaccharide (LPS) and to investigate the effect of Rg1 regulating proliferation in activated BV-2 cells. Methods BV-2 cells were divided into control group; LPS treatment groups with LPS stimulated for 1, 3, 6, 8, 12 hours, respectively and the Rg1 pretreatment groups in which the cells were pretreated with Rg1 (10, 20 and 50μmol/L) prior to LPS stimulate. The protein level and mRNA expression of PCNA and cyclin D1 were detected by RT-PCR and Western blotting, respectively. The expressions of cyclin D1 were detected by immunofluorescence staining. The cell cycle was detected by flow cytometry. Results The protein level and mRNA expression of PCNA and cyclin D1 in BV-2 cells were significantly increased following the treatment with LPS, which was in a time dependent manner. Compared with LPS group, addition of Rg1 inhibited the increase of PCNA and cyclin D1 in a dose-dependent manner. Conclusion Rg1 regulates the proliferation in activated BV-2 cells through inhibiting the expression of PCNA and cyclin D1.

Key words: [Key words] Rg1, LPS, Ginsenoside Rg1, Lipopolysaccharide, Proliferation；BV-2 cell, Proliferating cell nuclear antigen, Cyclin D1, Flow cytometry

姚玥伊艾青龙陈媛丽尚群竺张汝江毕秀梅孙俊钟莲梅. 人参皂苷Rg1抑制脂多糖诱导BV-2小胶质细胞增殖[J]. 解剖学报, 2016, 47(5): 614-619.

YAO Yue-yi AI Qing-long CHEN Yuan-li SHANG Qun-zhu ZHANG Ru-jiang BI Xiu-mei SUN Jun ZHONG Lian-mei. Effect of ginsenoside Rg1 on proliferation induced by lipopolysaccharide in BV-2 cells[J]. Acta Anatomica Sinica, 2016, 47(5): 614-619.

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人参皂苷Rg1抑制脂多糖诱导BV-2小胶质细胞增殖

Effect of ginsenoside Rg1 on proliferation induced by lipopolysaccharide in BV-2 cells

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