[1]薛鑫,陈星星,王冠,等.甲酰基肽受体1对BV-2细胞迁移的影响及机制的体外研究[J].陆军军医大学学报(原第三军医大学学报),2016,38(01):44-49.
 Xue Xin,Chen Xingxing,Wang Guan,et al.Formyl peptide receptor 1 promotes migration in mouse microglial cell line BV-2[J].J Amry Med Univ (J Third Mil Med Univ),2016,38(01):44-49.
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甲酰基肽受体1对BV-2细胞迁移的影响及机制的体外研究(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
38卷
期数:
2016年第01期
页码:
44-49
栏目:
基础医学
出版日期:
2016-01-15

文章信息/Info

Title:
Formyl peptide receptor 1 promotes migration in mouse microglial cell line BV-2
作者:
薛鑫陈星星王冠郭乔楠刘明永赵建华
第三军医大学大坪医院野战外科研究所脊柱外科;第三军医大学新桥医院病理科
Author(s):
Xue Xin Chen Xingxing Wang Guan Guo Qiaonan Liu Mingyong Zhao Jianhua

Department of Spine Surgery, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing, 400042; Department of Pathology, Xinqiao Hospital, Third Military Medical University, Chongqing,400037, China

关键词:
FPR1fMLPBoc-MLF细胞迁移小胶质细胞
Keywords:
formyl peptide receptor 1 formylmethionyl-leucyl-phenylalanine Boc-MLP migration ERK signal pathway
分类号:
R322.81; R329-33; R329.24
文献标志码:
A
摘要:

目的      探讨甲酰基肽受体1(formyl peptide receptor 1,FPR1)对BV-2细胞迁移的影响及潜在机制。      方法      免疫荧光染色检测BV-2细胞FPR1的表达,Transwell小室实验分别检测浓度为1、10、50、100、500 nmol/L的FPR1特异性激动剂(formylmethionyl-leucyl-phenylalanine,fMLP)对细胞迁移的影响,划痕实验及Transwell小室实验验证fMLP促进细胞迁移的作用可以被FPR1特异性阻断剂Boc-MLF所抑制。Western blot检测激动剂组、阻断剂组和阻断剂+激动剂组BV-2细胞经干预后其FPR1蛋白和磷酸化细胞外信号调节激酶(phospho-extracellular regulate kinase,p-ERK)蛋白表达量的变化。      结果      经免疫荧光染色结果显示,BV-2细胞表达FPR1,主要位于细胞膜表面。随着fMLP浓度的升高,Transwell迁移实验结果显示BV-2细胞的迁移能力明显增加(P<0.05),呈现出明显的剂量依赖性。100 nmol/L及500 nmol/L激动剂组效果最明显,fMLP促进BV-2细胞迁移的最适浓度为100 nmol/L。划痕及Transwell小室实验结果显示5 μmol/L的Boc-MLF可显著阻断上述现象(P<0.05)。fMLP可以上调BV-2细胞中FPR1的表达(P<0.05),并且显著活化胞内ERK信号通路,上调p-ERK蛋白的表达水平(P<0.05),Boc-MLF又可显著抑制上述改变(P<0.05)。      结论      BV-2细胞表达FPR1,且FPR1在促进BV-2细胞迁移中具有重要作用。

Abstract:

Objective      To determine the effect of formyl peptide receptor 1 (FPR1) on the migration of BV2 cells and investigate the underlying mechanism.       Methods       The expression of FPR1 receptor in BV-2 cells was detected by immunofluorescence staining. Transwell assay was used to detect the effect of FPR1 receptor specific agonist, formylmethionyl-leucyl-phenylalanine (fMLP), at different concentrations (1, 10, 50, 100 and 500 nmol/L) on the alterations of cell migration, while the scratch test and Transwell assay were used to verify the role of fMLP in the promotion of cell migration whether can be inhibited by FPR1 specific antagonist, Boc-MLF, or not. The protein expression of FPR1 was detected by Western blotting in the BV-2 cells stimulated with different concentrations of fMLP. After stimulated with 100 nmol/L fMLP and 5 μmol/L Boc-MLF, the expression of phospho-extracellular regulate kinase (p-ERK) in the cells was detected by Western blotting.       Results      Immunofluorescence staining indicated that FPR1 receptor was expressed in BV-2 cells, mainly located in the cell membrane. With the increase of fMLP doses, Transwell assay showed that the migration of BV-2 cells was enhanced significantly (P<0.05) in a dose-dependent fashion. The 100 and 500 nmol/L fMLP showed most obvious effect, while the optimal fMLP concentration to promote BV-2 cell migration was 100 nmol/L. The scratch test and Transwell assay verified that those above effects could be inhibited by 5 μmol/L Boc-MLF (P<0.05). The specific FPR1 agonist, fMLP up-regulated the FPR1 receptor expression in BV-2 cells (P<0.05), activated ERK signaling pathway, and increased p-ERK protein expression significantly (P<0.05). The FPR1 specific antagonist Boc-MLF inhibited the above changes significantly (P<0.05).       Conclusion      FPR1 receptor is expressed in the BV-2 cells, and FPR1 plays an important role in the promotion of BV-2 cells migration.

更新日期/Last Update: 2016-01-06