孙锦霞,王 瑞,谭 笑,等.2-BP对中性粒细胞趋化性的抑制作用[J].深圳大学学报理工版,2017,34(No.6(551-660)):625-630.[doi:10.3724/SP.J.1249.2017.06625]
Sun Jinxia,Wang Rui,Tan Xiao,et al.The inhibitory effect of 2-BP on neutrophil chemotaxis[J].Journal of Shenzhen University Science and Engineering,2017,34(No.6(551-660)):625-630.[doi:10.3724/SP.J.1249.2017.06625]
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2-BP对中性粒细胞趋化性的抑制作用
孙锦霞1,王 瑞2,谭 笑3,焦琛琛4,黄 钟1

1)深圳大学医学部中心实验室,广东深圳518060; 2)聊城大学药学院,山东聊城252059; 3)上海市第十人民医院中心实验室,上海200072; 4)同济大学医学院,上海200072

中性粒细胞; 2-溴棕榈酸; F-肌动蛋白极化; 趋化性; 免疫荧光; 流式细胞术; 免疫印迹; 细胞迁移; 磷脂酰肌醇3-激酶-蛋白激酶B信号通路

The inhibitory effect of 2-BP on neutrophil chemotaxis
Sun Jinxia1, Wang Rui2, Tan Xiao3, Jiao Chenchen4, and Huang Zhong1

Sun Jinxia1, Wang Rui2, Tan Xiao3, Jiao Chenchen4, and Huang Zhong11)Health Science Center, Shenzhen University, Shenzhen 518060, Guangdong Province, P.R.China2)College of Pharmacy, Liaocheng University, Liaocheng 252059, Shandong Province, P.R.China3)Central Laboratory of Shanghai Tenth People's Hospital, Shanghai 200072, P.R.China4)School of Medicine, Tongji University, Shanghai 200072, P.R.China

neutrophil; 2-bromopalmitate(2-BP); F-actin polarization; chemotaxis; immuno-fluorescence; flow cytometry; western blot; cell migration; phosphatidylinositol 3-kinase-AKT signaling pathway

DOI: 10.3724/SP.J.1249.2017.06625

备注

为研究棕榈酰化对中性粒细胞趋化性的调节作用,利用2-溴棕榈酸(2-bromopalmitate, 2-BP)或棕榈酸(palmitate)预处理来源于骨髓的中性粒细胞.采用流式细胞术(flow cytometry, FCM)检测中性粒细胞的存活率和中性粒细胞内F-肌动蛋白(F-actin)的生成; 通过免疫荧光分析中性粒细胞内F-actin的极化; 利用细胞迁移试验检测中性粒细胞的迁移能力; 通过免疫印迹的实验方法检测蛋白激酶B(protein kinase B,AKT)和磷酸化AKT的表达.结果发现,2-BP预处理显著抑制了中性粒细胞内F-actin极化,且抑制了中性粒细胞向趋化肽N-甲酰甲硫氨酰-亮氨酰-苯丙氨酸(N-formylmethionyl-leucyl-phenylalanine,fMLP)迁移的能力,而棕榈酸预处理显著促进了中性粒细胞内F-肌动蛋白极化.两者对F-actin生成和AKT的磷酸化均无影响.研究结果表明,2-BP处理可能通过影响F-actin的极化抑制了中性粒细胞向fMLP的趋化活性.

Bone marrow derived neutrophil was pretreated with 2-BP or palmitate in order to explore regulatory roles of palmitoylation on neutrophil chemotaxis. Flow cytometry(FCM)was used to detect viability and F-actin formation of neutrophil. Polarization of F-actin in neutrophil was analyzed via immune-fluorescence(IF)analysis. Cell migration assay was performed to analyz migration ability of neutrophil. Expression of protein kinase B(AKT)and phosphorylated AKT was tested through Western Blot(WB). The results indicate that neutrophil pretreated with 2-BP display significantly reduces F-actin polarization and poor ability of migration toward chemotactic peptide N-formylmethionyl-leucyl-phenylalanine(fMLP). However, palmitate pretreatment obviously increases polarization of F-actin in neutrophil. Furthermore, both 2-BP and palmitate have no effects on F-actin formation and AKT phosphorylation. Thus, these data suggest that 2-BP pretreatment inhibition on neutrophil chemotaxis toward fMLP may be through regulating F-actin polarization.

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