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学科主题: 免疫学
题名:
白细胞介素-21对 RAW264.7细胞向破骨细胞分化的影响
其他题名: Effect of interleukin-21 on osteoclastogenesis in RAW264.7 cells
作者: 张颖健; 李常虹; 邢瑞; 孙琳; 赵金霞; 刘湘源
关键词: 白细胞介素-21 ; 破骨细胞 ; RAW264畅7 ; Interleukin-21 ; Osteoclast ; RAW264.7
刊名: 中国骨质疏松杂志
发表日期: 2015
期: 3, 页:253-258
收录类别: 中国科技核心期刊 ; 中文核心期刊 ; CSCD
文章类型: Journal Article
摘要: 目的:了解白细胞介素-21( IL-21)诱导小鼠巨噬细胞系RAW264.7细胞向破骨细胞分化的作用,并探讨其可能的机制。方法1、以不同浓度IL-21(0,1,10,20,40 ng/ml)处理RAW264.7细胞,培养5天后进行抗酒石酸酸性磷酸酶( TRAP)染色,Western Blot和免疫组化法检测降钙素受体(CTR)表达,采用Real time PCR 法检测CTR和组织蛋白酶(Cathepsin)-K的mRNA表达水平。2、以信号通路抑制剂AG490、LY294002和PD98059作用30min后再加入IL-21,培养5天后观察破骨细胞形成情况,以Western Blot检测IL-21作用不同时间点时信号通路分子总蛋白和磷酸化蛋白水平,探讨IL-21直接诱导破骨细胞分化的作用机制。结果1、在无RANKL作用的情况下,随着IL-21浓度增加TRAP阳性细胞数逐渐增多,20 ng/ml作用最强,40ng/ml时减弱。 IL-21能够诱导破骨标志分子CTR和Cathepsin-K mRNA水平表达上调。 Western Blot和免疫细胞化学证实,与阴性组比较,IL-21能促进 CTR 蛋白水平表达增高。2、PI3K-AKT 通路抑制剂( LY294002)可以显著抑制 IL-21诱导的RAW264.7细胞向破骨细胞分化,IL-21刺激5~15 min时p-AKT表达增强。结论 IL-21可不依赖RANKL直接诱导小鼠巨噬细胞系RAW264.7细胞向破骨细胞分化,该作用可能由PI3K-AKT通路介导。 Objective To investigate the role of IL-21 in osteoclastogenesis of RAW264.7 cells and its possible mechanism. Methods 1) RAW264.7 cells were treated with different concentrations of IL-21 (0, 1, 10, 20, and 40 ng/ml).After 5 days, TRAP staining was performed. The calcitonin receptor ( CTR ) expression was detected using Western blot and immunocytochemistry.The mRNA levels of CTR and Cathepsin-K were examined using real-time PCR.2 ) Before adding IL-21 into the culture, various signaling pathway inhibitors ( AG490, LY294002 and PD98059) were incubated with RAW264.7 for 30 min.Osteoclast formation was determined after 5 days culture.Phosphorylation of signaling proteins was determined using Western blot under the stimulation of IL-21.Results 1) Without the presence of RANKL, TRAP positive cells increased with the increase of IL-21 concentration.The strongest effect was at the 20 ng/ml concentration, and the effect decreased at the 40 ng/ml concentration.IL-21 promoted mRNA expression of CTR and Cathepsin-K.Western blot and immunocytochemistry showed that the expression of CTR in IL-21 group increased compared to that in control group.2) The PI3K-AKT pathway inhibitor LY294002 significantly suppressed IL-21-induced osteoclastogenesis in RAW264.7 cells. P-AKT expression increased after 5-15 min stimulation of IL-21.Conclusion IL-21 can induce osteoclastogenesis of RAW264.7 cells independent of RANKL via PI3K-AKT pathway.
语种: 中文
原文出处: 查看原文
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内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/69682
Appears in Collections:北京大学第三临床医学院_风湿免疫科_期刊论文

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作者单位: 1.北京大学第三医院风湿免疫科,北京 100191
2.北京市公安局强制治疗管理处
3.北京大学第三医院风湿免疫科,北京,100191

Recommended Citation:
张颖健,李常虹,邢瑞,等. 白细胞介素-21对 RAW264.7细胞向破骨细胞分化的影响[J]. 中国骨质疏松杂志,2015(3):253-258.
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