2-BP對(duì)中性粒細(xì)胞趨化性的抑制作用

孫錦霞，王 瑞，譚 笑，焦琛琛，黃 鐘

1)深圳大學(xué)醫(yī)學(xué)部中心實(shí)驗(yàn)室，廣東深圳518060；2)聊城大學(xué)藥學(xué)院，山東聊城252059；3)上海市第十人民醫(yī)院中心實(shí)驗(yàn)室，上海200072；4)同濟(jì)大學(xué)醫(yī)學(xué)院，上海200072

【生物工程/Bioengineering】

2-BP對(duì)中性粒細(xì)胞趨化性的抑制作用

孫錦霞1，王 瑞2，譚 笑3，焦琛琛4，黃 鐘1

1)深圳大學(xué)醫(yī)學(xué)部中心實(shí)驗(yàn)室，廣東深圳518060；2)聊城大學(xué)藥學(xué)院，山東聊城252059；3)上海市第十人民醫(yī)院中心實(shí)驗(yàn)室，上海200072；4)同濟(jì)大學(xué)醫(yī)學(xué)院，上海200072

為研究棕櫚酰化對(duì)中性粒細(xì)胞趨化性的調(diào)節(jié)作用，利用2-溴棕櫚酸(2-bromopalmitate, 2-BP)或棕櫚酸(palmitate)預(yù)處理來源于骨髓的中性粒細(xì)胞．采用流式細(xì)胞術(shù)(flow cytometry, FCM)檢測(cè)中性粒細(xì)胞的存活率和中性粒細(xì)胞內(nèi)F-肌動(dòng)蛋白(F-actin)的生成；通過免疫熒光分析中性粒細(xì)胞內(nèi)F-actin的極化；利用細(xì)胞遷移試驗(yàn)檢測(cè)中性粒細(xì)胞的遷移能力；通過免疫印跡的實(shí)驗(yàn)方法檢測(cè)蛋白激酶B(protein kinase B，AKT)和磷酸化AKT的表達(dá)．結(jié)果發(fā)現(xiàn)，2-BP預(yù)處理顯著抑制了中性粒細(xì)胞內(nèi)F-actin極化，且抑制了中性粒細(xì)胞向趨化肽N-甲酰甲硫氨酰-亮氨酰-苯丙氨酸(N-formylmethionyl-leucyl-phenylalanine，fMLP)遷移的能力，而棕櫚酸預(yù)處理顯著促進(jìn)了中性粒細(xì)胞內(nèi)F-肌動(dòng)蛋白極化．兩者對(duì)F-actin生成和AKT的磷酸化均無(wú)影響．研究結(jié)果表明，2-BP處理可能通過影響F-actin的極化抑制了中性粒細(xì)胞向fMLP的趨化活性．

中性粒細(xì)胞；2-溴棕櫚酸；F-肌動(dòng)蛋白極化；趨化性；免疫熒光；流式細(xì)胞術(shù)；免疫印跡；細(xì)胞遷移；磷脂酰肌醇3-激酶-蛋白激酶B信號(hào)通路

中性粒細(xì)胞是外周血中含量最多的白細(xì)胞，可參與調(diào)節(jié)急性以及一些慢性炎癥反應(yīng)進(jìn)程. 其源自骨髓造血干細(xì)胞，壽命較短，在粒細(xì)胞集落刺激因子( granulocyte colony-stimulating factor，G-CSF)和粒細(xì)胞-巨噬細(xì)胞集落刺激因子(granulocyte-macrophage CSF，GM-CSF)的作用下，可從骨髓中釋放出來，進(jìn)入循環(huán)系統(tǒng). 靜息狀態(tài)下，中性粒細(xì)胞在循環(huán)系統(tǒng)中約存活6 h，隨后被肝臟、脾臟或骨髓清除[1-3]．當(dāng)感染發(fā)生時(shí)，中性粒細(xì)胞被快速且選擇性的動(dòng)員起來，以確保在感染部位大量聚集. 一旦中性粒細(xì)胞到達(dá)目的地，它們的短暫壽命也得到了延長(zhǎng)[4-5].作為一線免疫細(xì)胞，中性粒細(xì)胞可快速穿過血管內(nèi)皮細(xì)胞到達(dá)感染部位，通過吞噬、脫顆粒和中性粒細(xì)胞胞外網(wǎng)絡(luò)(neutrophil extracellular traps, NET)清除病原菌，隨后凋亡，被巨噬細(xì)胞清除以控制炎癥進(jìn)程[6-10].長(zhǎng)期以來，科學(xué)家們認(rèn)為吞噬和脫顆粒是中性粒細(xì)胞抵抗病原菌感染的唯一機(jī)制，直至中性粒NET的發(fā)現(xiàn)，科學(xué)家們意識(shí)到這樣一個(gè)問題，針對(duì)不同的病原菌感染，中性粒細(xì)胞是否可以選擇性殺傷病原菌，從而激發(fā)最優(yōu)的免疫反應(yīng)[11-12]．

趨化性是中性粒細(xì)胞得以遷移的生物學(xué)基礎(chǔ)，包括對(duì)趨化物濃度梯度的感應(yīng)、細(xì)胞極化和定向遷移的過程，首先F-actin發(fā)生極化并在細(xì)胞前端行成偽足，后端行成腹足，F(xiàn)-actin的持續(xù)生成使得中性粒細(xì)胞得以向著趨化物濃度梯度發(fā)生定向遷移[13].研究表明，抑癌蛋白PTEN(phosphatase and tensin homolog)[14]、PI3K-AKT[15]信號(hào)通路、小G蛋白(Cdc42、Rac和RhoA)[15]等在細(xì)胞極化和遷移過程中發(fā)揮至關(guān)重要的調(diào)節(jié)作用.

櫚酰化修飾是一種可逆的共價(jià)脂肪酸化修飾，指16碳棕櫚酸(palmitate)通過不穩(wěn)定硫酯鍵結(jié)合至細(xì)胞質(zhì)特定的半胱氨酸殘基側(cè)鏈上. 研究表明，棕櫚?；揎椏烧{(diào)節(jié)蛋白質(zhì)功能的所有階段，包括蛋白質(zhì)的成熟加工、胞內(nèi)運(yùn)輸、細(xì)胞器定位、蛋白質(zhì)穩(wěn)定性以及與其他蛋白的相互作用[16-18]．因此，棕櫚?；揎棇?duì)蛋白質(zhì)功能的正常發(fā)揮、突觸的可塑性和免疫調(diào)節(jié)等多種生物學(xué)過程的影響至關(guān)重要[19-20]．棕櫚酰化和脫棕櫚?；^程分別由蛋白質(zhì)?；D(zhuǎn)移酶和?；鞍琢蝓ッ复呋痆21-22]．2-BP通過抑制棕櫚酰?；D(zhuǎn)移酶的活性，不可逆地抑制蛋白質(zhì)發(fā)生棕櫚?；揎梉23]. 本研究就2-BP和palmitate預(yù)處理對(duì)中性粒細(xì)胞內(nèi)F-actin極化、 F-actin生成、細(xì)胞遷移和AKT磷酸化(phosphorylated AKT, p-AKT)的作用展開研究，以闡明2-BP對(duì)中性粒細(xì)胞趨化性的調(diào)節(jié)作用及其潛在調(diào)控機(jī)制.

1 材料與方法

1.1 主要試劑與儀器

Hank’s平衡鹽緩沖液(Hank’s balanced salt solution，HBSS) 緩沖液購(gòu)自美國(guó)Gibco公司；NH4Cl、KHCO3、乙二胺四乙酸(ethylene diamine tetraacetic acid，EDTA)和體積分?jǐn)?shù)為4%的多聚甲醛等化學(xué)試劑購(gòu)自上海生工生物工程股份有限公司；Percoll細(xì)胞分離液購(gòu)自美國(guó)GE Healthcare公司；二甲基亞砜(dimethyl sulfoxide, DMSO)、2-BP、palmitate、fMLP和溶血卵磷脂均購(gòu)自美國(guó)Sigma公司，鬼筆環(huán)肽購(gòu)自美國(guó)Invitrogen公司；染料DAPI(2-(4-amidinophenyl)-6-indolecarbamidine dihydrochloride)購(gòu)自上海碧云天生物技術(shù)有限公司；碘化丙啶-膜聯(lián)蛋白V(propidium iodide-annexin V，PI-Annexin V)凋亡檢測(cè)試劑盒購(gòu)自美國(guó) BD公司；Trans-well購(gòu)自美國(guó)Corning公司；抗p-AKT、抗AKT抗體和熒光標(biāo)記二抗購(gòu)自美國(guó)CST公司；倒置顯微鏡購(gòu)自日本Olympus公司；激光共聚焦顯微鏡購(gòu)自德國(guó)Leica公司；高速冷凍離心機(jī)購(gòu)自美國(guó)Beckman公司；流式細(xì)胞儀購(gòu)自美國(guó)BD公司；Odyssey雙色紅外成像系統(tǒng)購(gòu)自美國(guó)LI-COR公司.

1.2 中性粒細(xì)胞提取

8～12周C57BL/6小鼠處死后，取股骨和脛骨，用10 mL HBSS 緩沖液(無(wú)Ca2+和Mg2+)(含有10 g/L牛血清白蛋白(bovine serum albumin，BSA)和5 mmol/L Hepes)沖出骨髓細(xì)胞. 經(jīng)紅細(xì)胞裂解液(含有0.15 mol/L NH4Cl、10.0 mmol/L KHCO3和0.1 mmol/L EDTA)裂解紅細(xì)胞后，加入3 mL體積分?jǐn)?shù)為45%的Percoll細(xì)胞分離液重懸骨髓細(xì)胞，置于冰上. 在15 mL離心管中依次加入3 mL體積分?jǐn)?shù)分別為81%和62%的Percoll及細(xì)胞懸液. 密度梯度離心30 min后，吸取體積分?jǐn)?shù)為81%和62% 的Percoll中間細(xì)胞層，即成熟中性粒細(xì)胞.

1.3 免疫熒光

HBSS 緩沖液(含Ca2+和Mg2+)稀釋提取的中性粒細(xì)胞至1×106mL-1，每孔20 μL加至鋪有載玻片的24孔細(xì)胞培養(yǎng)板，37 ℃貼壁5 min. 加入等體積濃度為20 μmol/L的N-甲酰甲硫氨酰-亮氨酰-苯丙氨酸(N-formylmethionyl-leucyl-phenylalanine，fMLP)刺激不同時(shí)間，迅速加入體積分?jǐn)?shù)為4%多聚甲醛固定20 min. PBS洗滌后加入體積分?jǐn)?shù)為0.1% TritonX-100孵育5 min. PBS洗滌后加入鬼筆環(huán)肽染色F-actin 30 min. PBS洗滌后加入DAPI染色細(xì)胞核5 min，洗后封片，利用激光共聚焦顯微鏡采集圖片.

1.4 流式細(xì)胞術(shù)

用HBSS 緩沖液(含Ca2+和Mg2+)稀釋提取的中性粒細(xì)胞至1×107mL-1，設(shè)7個(gè)組，分別取100 μL細(xì)胞懸液，做如下預(yù)處理，組號(hào)分別為：① 加入終濃度為1 μmol/L的2-BP；② 加入終濃度為10 μmol/L的2-BP；③ 加入終濃度為20 μmol/L的2-BP；④ 加入終濃度為40 μmol/L的2-BP；⑤ 加入終濃度為50 μmol/L的2-BP；⑥ 加入體積分?jǐn)?shù)為0.1%的DMSO；⑦ 加入終濃度為50 μmol/L的palmitate.將7組細(xì)胞分別放入37 ℃孵育2 h，然后按照PI/Annexin V凋亡檢測(cè)試劑盒說明書操作，加入PI和異硫氰酸熒光素(fluorescein isothiocyanate, FITC)標(biāo)記的AnnexinV染色15 min，通過流式細(xì)胞術(shù)檢測(cè)各組樣品中細(xì)胞存活率的差異.

另取100 μL細(xì)胞懸液，加入終濃度為20 μmol/L的palmitate，記做預(yù)處理⑧，分別將預(yù)處理③、⑥和⑧組細(xì)胞于37 ℃孵育2 h，經(jīng)PBS洗滌后用10 μmol/L fMLP刺激不同時(shí)間，加入體積分?jǐn)?shù)為4%多聚甲醛固定20 min，PBS洗滌后加入20 mg/mL溶血卵磷脂和鬼筆環(huán)肽染色F-actin 30 min，流式細(xì)胞術(shù)檢測(cè)不同處理后F-actin生成的差異.

1.5 細(xì)胞遷移

分別將預(yù)處理的③、⑥和⑧組細(xì)胞于37 ℃孵育2 h，然后取20 μL細(xì)胞加入96孔Trans-well上室，下室加入60 μL的10 μmol/L fMLP，37 ℃培養(yǎng)1 h，倒置顯微鏡下隨機(jī)采集5個(gè)視野，細(xì)胞計(jì)數(shù)后統(tǒng)計(jì)不同處理后細(xì)胞遷移的差異.

1.6 免疫印跡

HBSS 緩沖液(含Ca2+和Mg2+)稀釋提取的中性粒細(xì)胞至1×108mL-1，分別將預(yù)處理③、⑥和⑧組細(xì)胞于37 ℃孵育2 h．經(jīng)PBS洗滌后，于37 ℃用10 μmol/L fMLP分別刺激不同時(shí)間，加入SDS上樣緩沖液終止反應(yīng)并裂解細(xì)胞. 樣品于100 ℃煮沸10 min，用于十二烷基硫酸鈉聚丙烯酰胺凝膠電泳(sodium dodecyl sulfate polyacrylamide gel electropheresis, SDS-PAGE)凝膠電泳檢測(cè). 抗p-AKT和抗AKT一抗于4 ℃孵育過夜，熒光標(biāo)記的二抗室溫孵育1 h，使用Odyssey雙色紅外成像系統(tǒng)采集圖片.

1.7 統(tǒng)計(jì)學(xué)方法

2 結(jié)果與分析

2.1 不同濃度2-BP對(duì)中性粒細(xì)胞存活率的影響

為檢測(cè)棕櫚?；瘜?duì)中性粒細(xì)胞趨化性的調(diào)控作用，本研究先檢測(cè)了2-BP和palmitate對(duì)其存活率的影響．將流式細(xì)胞術(shù)處理后的①～⑦組細(xì)胞分別進(jìn)行檢測(cè)，結(jié)果如圖1．從圖1中可見，與對(duì)照組DMSO相比，當(dāng)2-BP的濃度增加至40～50 μmol/L時(shí)可以促進(jìn)細(xì)胞凋亡，而濃度為50 μmol/L的palmitate對(duì)細(xì)胞存活率仍沒有影響. 因此，后續(xù)實(shí)驗(yàn)將選用濃度為20 μmol/L 的2-BP和palmitate的預(yù)處理方式.

圖1 2-BP、palmitate和DMSO對(duì)中性粒細(xì)胞存活率的影響Fig.1 Effects of f 2-BP, palmitate and DMSO on neutrophil viability

2.2 2-BP對(duì)中性粒細(xì)胞遷移的抑制作用

將1.4節(jié)預(yù)處理的③、⑥和⑧組細(xì)胞分別加入Trans-well上室，于37 ℃向含有fMLP的下室定向遷移1 h，隨后通過后倒置顯微鏡采集圖片，分析2-BP和palmitate對(duì)中性粒細(xì)胞遷移的影響，結(jié)果如圖2．由圖2可見，20 μmol/L 2-BP顯著抑制中性粒細(xì)胞向fMLP的遷移，而20 μmol/L palmitate對(duì)中性粒細(xì)胞向fMLP的遷移則沒有影響.

2.3 2-BP對(duì)中性粒細(xì)胞內(nèi)F-actin極化和生成的調(diào)節(jié)作用

將預(yù)處理③、⑥和⑧組細(xì)胞于37 ℃孵育2 h，然后分作2組，1組分別加入10 μmol/L fMLP分別刺激0、1和3 min，固定后依次染色鬼筆環(huán)肽和DAPI，激光共聚焦顯微鏡檢測(cè)F-actin極化的差異，結(jié)果如圖3；另1組分別加入10 μmol/L fMLP分別刺激0、1和3 min，固定后加入20 mg/mL溶血卵磷脂和鬼筆環(huán)肽孵育30 min，流式細(xì)胞術(shù)檢測(cè)F-actin生成的差異，結(jié)果如圖4． 由圖3可見， 2-BP預(yù)處理顯著抑制fMLP介導(dǎo)的F-actin在偽足極化，palmitate則促進(jìn)了fMLP介導(dǎo)的F-actin在偽足極化. 由圖4可見，2-BP和palmitate預(yù)處理對(duì)F-actin的生成均無(wú)影響.

圖2 2-BP、palmitat和DMSO對(duì)中性粒細(xì)胞向fMLP遷移能力的調(diào)節(jié)作用Fig.2 Regulation of 2-BP, palmitate and DMSO on neutrophil migration toward fMLP

圖3 2-BP、palmitate和DMSO對(duì)中性粒細(xì)胞內(nèi)F-actin極化的調(diào)節(jié)作用Fig.3 Regulation of 2-BP, palmitate and DMSO on F-actin polarization in neutrophil

圖4 2-BP、palmitate和DMSO對(duì)中性粒細(xì)胞內(nèi)F-acin生成的調(diào)節(jié)作用Fig.4 Regulation of 2-BP, palmitate and DMSO on F-actin formation in neutrophil

2.4 2-BP對(duì)AKT磷酸化的調(diào)節(jié)作用

分別將預(yù)處理③、⑥和⑧組細(xì)胞于37 ℃孵育2 h，然后加入10 μmol/L fMLP分別刺激0、1和3 min，加入SDS上樣緩沖液終止反應(yīng)并裂解細(xì)胞，樣品用于SDS-PAGE凝膠電泳，Odyssey雙色紅外成像系統(tǒng)檢測(cè)p-AKT和AKT表達(dá)的差異，結(jié)果如圖5．由圖5可見，2-BP和palmitate預(yù)處理對(duì)AKT的磷酸化均無(wú)影響.

圖5 2-BP、palmitate和DMSO對(duì)中性粒細(xì)胞內(nèi)AKT磷酸化的調(diào)節(jié)作用Fig.5 Effects of 2-BP, palmitate and DMSO on AKT phosphorylation in neutrophil

3 討 論

棕櫚?；怯蓹磅ｕ；D(zhuǎn)移酶催化發(fā)生的一種動(dòng)態(tài)翻譯后修飾，1979年首次發(fā)現(xiàn)于水泡性口膜炎病毒跨膜糖蛋白[24]. 棕櫚酰酰基轉(zhuǎn)移酶為高度保守天冬氨酸-組氨酸-組氨酸-半胱氨酸(Asp-His-His-Cys, DHHC)家族蛋白，其中哺乳細(xì)胞內(nèi)有23種[25]. 與豆蔻?；彤愇煜；嗨?，櫚?；揎椧部梢栽黾拥鞍踪|(zhì)疏水性，介導(dǎo)靶蛋白定位于細(xì)胞膜，增加靶蛋白與細(xì)胞膜的結(jié)合進(jìn)而調(diào)節(jié)蛋白的功能[26]. 迄今尚無(wú)棕櫚?；c中性粒細(xì)胞趨化性的報(bào)道，本研究采用2-BP和palmitate分別預(yù)處理中性粒細(xì)胞，首次證明2-BP顯著抑制中性粒細(xì)胞內(nèi)F-acin的極化且不影響其生成，Trans-well實(shí)驗(yàn)結(jié)果顯示，2-BP顯著抑制了中性粒細(xì)胞向fMLP的遷移. 因此，2-BP可能通過抑制F-acin極化參與調(diào)節(jié)中性粒細(xì)胞的趨化性. 此外，本研究還發(fā)現(xiàn)隨著2-BP濃度的增高，可以不同程度的促進(jìn)中性粒細(xì)胞凋亡，但其促凋亡機(jī)制尚不清楚．

中性粒細(xì)胞是先天免疫和急性炎癥反應(yīng)的關(guān)鍵效應(yīng)細(xì)胞，它們?cè)谕鈬M織中的招募是機(jī)體發(fā)揮宿主防御功能必不可少的.由于它們具有潛在的破壞性，因此中性粒細(xì)胞進(jìn)入外周組織必須受到嚴(yán)格的控制，以避免對(duì)機(jī)體造成不必要的傷害[27].趨化因子、白三烯、補(bǔ)體、炎性因子及細(xì)菌釋放的多肽(如fMLP)都可以趨化中性粒細(xì)胞快速穿過血管壁到達(dá)感染部位[9]. palmitate與AKT信號(hào)通路的關(guān)系已有報(bào)道，如palmitate通過抑制PIP3K/AKT通路抑制內(nèi)皮細(xì)胞介導(dǎo)的血管生成[28]．但在中性粒細(xì)胞中，兩者關(guān)系未知．研究表明，阻斷PIP3K的激酶活性顯著抑制中性粒細(xì)胞在體內(nèi)外的遷移和向感染部位的募集，且促進(jìn)了中性粒細(xì)胞的程序性死亡[29]. 此外，Rho家族小GTPase在中性粒細(xì)胞內(nèi)極性分布，參與調(diào)節(jié)F-actin的定位和生成[15]. 本研究結(jié)果顯示，2-BP和palmitate均對(duì)PI3K/AKT信號(hào)通路的活化沒有影響，提示2-BP對(duì)中性粒細(xì)胞趨化性的抑制作用可能通過其他調(diào)控機(jī)制，如小GTPase等.

結(jié) 語(yǔ)

中性粒細(xì)胞參與調(diào)節(jié)宿主防御、腫瘤免疫和創(chuàng)傷修復(fù)等生理、病理進(jìn)程，其中趨化性是中性粒細(xì)胞快速募集至感染部位行使殺傷作用所必需的，研究表明，2-BP顯著抑制中性粒細(xì)胞的F-actin極化和遷移，提示棕櫚?；揎椏蓞⑴c調(diào)節(jié)在中性粒細(xì)胞的趨化性，其調(diào)節(jié)機(jī)制尚待后續(xù)深入研究.

引文：孫錦霞，王 瑞，譚 笑，等. 2-BP對(duì)中性粒細(xì)胞趨化性的抑制作用[J]. 深圳大學(xué)學(xué)報(bào)理工版，2017，34(6)：625-630.

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【中文責(zé)編：晨兮；英文責(zé)編：艾琳】

2016-11-14；Revised2017-09-12；Accepted2017-09-22

Professor Huang Zhong. E-mail: zhuang809@126.com

Theinhibitoryeffectof2-BPonneutrophilchemotaxis

SunJinxia1,WangRui2,TanXiao3,JiaoChenchen4,andHuangZhong1

1) 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

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 peptideN-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.

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

Foundation：National Natural Science Foundation of China (31401217); Postdoctral Science Foundation of China (2014M0560672); Shenzhen Science and Technology Basic Research Foundation (JCYJ20150403091443312)

：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(6): 625-630.(in Chinese)

R 392

A

10.3724/SP.J.1249.2017.06625

國(guó)家自然科學(xué)基金資助項(xiàng)目(31401217)；中國(guó)博士后科學(xué)基金資助項(xiàng)目(2014M0560672); 深圳市科技基礎(chǔ)研究計(jì)劃資助項(xiàng)目(JCYJ20150403091443312)

孫錦霞(1986—)，女，深圳大學(xué)博士后研究人員. 研究方向：免疫學(xué). E-mail：jinxia8608@126.com