体育科学
體育科學
체육과학
China Sport Science
2011年
3期
37~43
,共null页
董静梅 陈佩杰 王茹 于动震 张亚军 肖卫华
董靜梅 陳珮傑 王茹 于動震 張亞軍 肖衛華
동정매 진패걸 왕여 우동진 장아군 초위화
乳酸 吞噬细胞 吞噬功能 呼吸爆发 活性氧 NADPH氧化酶
乳痠 吞噬細胞 吞噬功能 呼吸爆髮 活性氧 NADPH氧化酶
유산 탄서세포 탄서공능 호흡폭발 활성양 NADPH양화매
lactic acid; phagocytic cells; phagocytosis function; respiratory burst; radical; ROS; NADPH-oxidase
目的：探讨乳酸引起的酸度变化对外周全血吞噬细胞呼吸爆发与吞噬功能的影响及发生机制。方法：1）取20名健康男性外周静脉血10 ml,分10份建立乳酸体积量与血乳酸值的回归方程;2）另取10名健康男性外周静脉血20 ml分为对照组（Ctr组）、乳酸处理组（E组）,各组再分10小组,按照第一步建立的回归方程加入不同体积的乳酸,同时Ctr组给予DPI处理,2组同时孵育10 min后各小组每份取100μl全血用流式细胞仪测定呼吸爆发和吞噬功能;3）分别取Ctr组、E组第6小组的血液分离中性粒细胞,利用免疫细胞化学与共聚焦技术进行NADPH氧化酶关键亚基gp91 phox与p47 phox的共定位实验。结果：1）低浓度（低于4.81 mmol/L）的乳酸抑制吞噬细胞的呼吸爆发和吞噬功能,中度浓度（4.81 mmol/L~8.14 mmol/L）的乳酸浓度可引起吞噬细胞呼吸爆发依赖乳酸浓度的增加而增加,高于此浓度可引起吞噬细胞呼吸爆发与急剧下降,而吞噬功能提前急剧下降;2）吞噬细胞NADPH氧化酶活性的改变与呼吸爆发的变化呈高度的相关性;3）乳酸浓度为8.14 mmol/L时中性粒细胞NADPH氧化酶关键亚基p47 phox移为至质膜与gp91 phox出现共定位。结论：乳酸引起的外周血酸度的变化可以通过调控NADPH氧化酶活性的改变影响其吞噬细胞呼吸爆发和吞噬功能。
目的：探討乳痠引起的痠度變化對外週全血吞噬細胞呼吸爆髮與吞噬功能的影響及髮生機製。方法：1）取20名健康男性外週靜脈血10 ml,分10份建立乳痠體積量與血乳痠值的迴歸方程;2）另取10名健康男性外週靜脈血20 ml分為對照組（Ctr組）、乳痠處理組（E組）,各組再分10小組,按照第一步建立的迴歸方程加入不同體積的乳痠,同時Ctr組給予DPI處理,2組同時孵育10 min後各小組每份取100μl全血用流式細胞儀測定呼吸爆髮和吞噬功能;3）分彆取Ctr組、E組第6小組的血液分離中性粒細胞,利用免疫細胞化學與共聚焦技術進行NADPH氧化酶關鍵亞基gp91 phox與p47 phox的共定位實驗。結果：1）低濃度（低于4.81 mmol/L）的乳痠抑製吞噬細胞的呼吸爆髮和吞噬功能,中度濃度（4.81 mmol/L~8.14 mmol/L）的乳痠濃度可引起吞噬細胞呼吸爆髮依賴乳痠濃度的增加而增加,高于此濃度可引起吞噬細胞呼吸爆髮與急劇下降,而吞噬功能提前急劇下降;2）吞噬細胞NADPH氧化酶活性的改變與呼吸爆髮的變化呈高度的相關性;3）乳痠濃度為8.14 mmol/L時中性粒細胞NADPH氧化酶關鍵亞基p47 phox移為至質膜與gp91 phox齣現共定位。結論：乳痠引起的外週血痠度的變化可以通過調控NADPH氧化酶活性的改變影響其吞噬細胞呼吸爆髮和吞噬功能。
목적：탐토유산인기적산도변화대외주전혈탄서세포호흡폭발여탄서공능적영향급발생궤제。방법：1）취20명건강남성외주정맥혈10 ml,분10빈건립유산체적량여혈유산치적회귀방정;2）령취10명건강남성외주정맥혈20 ml분위대조조（Ctr조）、유산처리조（E조）,각조재분10소조,안조제일보건립적회귀방정가입불동체적적유산,동시Ctr조급여DPI처리,2조동시부육10 min후각소조매빈취100μl전혈용류식세포의측정호흡폭발화탄서공능;3）분별취Ctr조、E조제6소조적혈액분리중성립세포,이용면역세포화학여공취초기술진행NADPH양화매관건아기gp91 phox여p47 phox적공정위실험。결과：1）저농도（저우4.81 mmol/L）적유산억제탄서세포적호흡폭발화탄서공능,중도농도（4.81 mmol/L~8.14 mmol/L）적유산농도가인기탄서세포호흡폭발의뢰유산농도적증가이증가,고우차농도가인기탄서세포호흡폭발여급극하강,이탄서공능제전급극하강;2）탄서세포NADPH양화매활성적개변여호흡폭발적변화정고도적상관성;3）유산농도위8.14 mmol/L시중성립세포NADPH양화매관건아기p47 phox이위지질막여gp91 phox출현공정위。결론：유산인기적외주혈산도적변화가이통과조공NADPH양화매활성적개변영향기탄서세포호흡폭발화탄서공능。
Objective：To investigate the impact of acidity change caused by lactic acid on the phagocytes cell respiratory burst and the phagocytic function in peripheral whole blood and mechanism.Method：1） 10ml peripheral whole blood from 20 healthy male,was divided into 10 portions（1ml per portion） to establish the regression equation between concentration land the volume lactic acid.2） 20ml peripheral whole blood from another 10 healthy male,was divided into group Ctr and E,each group was further separated into 10 portions and added in the different volume of lactic acid according to the first step,the group Ctr was treated with DPI additionally.The blood of two groups was incubated for 10 minutes and then 100ul whole blood was taken from each portion to detect the function of respiratory burst and phagocytosis of neutrophils and mononuclear cell respectively by flow cytometery.3.The neutrophils from the sixth portion blood of group Ctr and E were taken and observed the co-localization of the gp91phox and p47phox in NADPH-oxidase using immunocytochemistry and confocal technology.Result：1） Low concentration（less than 4.81mmol / L） of lactic acid can inhibit the function of phagocytosis and respiratory burst of phagocytic cells,moderate concentration（4.81mmol / L~8.14mmol / L） of lactic acid can lead to the increase respiratory burst of phagocytic cells.The curve of respiratory burst can be declined sharply while above this concentration lactic acid and the curve of sharp decline of the function of phagocytosis was early than the curve of respiratory burst.2） It is a high degree of correlation between the activity NADPH oxidase and the respiratory burst in phagocytic cells.3） There is co-localization between the subunit p47phox and the gp91phox in NADPH-oxidase on the plasma membrane in neutrophils when the lactic acid concentration in blood is 8.14mmol / L.Conclusion：The acidity induced by lactic acid in peripheral blood can regulate the activity NADPH oxidase and lead to the change of the phagocytosis function and respiratory burst in phagocytic cells.
