高碳酸因素对大鼠活化T淋巴细胞CD28/B7和CD2/CD48(LFA-3)信号转导通路的影响  

Effects of high carbonate factors on CD28/B7 and CD2 / CD48( LFA-3) signal transduction pathways in T lymphocytes in rats

作　　者：张丽娟[1,2] 车玉香 高伟[1,2] 崔晓光 

机构地区：[1]哈尔滨医科大学附属第二医院麻醉科 [2]黑龙江省麻醉与危重病学研究重点实验室,黑龙江哈尔滨150081

出　　处：《哈尔滨医科大学学报》2014年第6期460-463,共4页Journal of Harbin Medical University

基　　金：黑龙江省教育厅科学技术研究项目(12521221)

摘　　要：目的：探讨高碳酸因素对大鼠活化T淋巴细胞CD28／B7和CD2／CD48（LFA－3）信号转导通路的影响。方法采集体重200～250 g的健康Wistar大鼠外周静脉血样，分离T淋巴细胞，培养于含有植物血凝素（ PHA）的无菌16孔培养板中。采用随机数字表法，将其分为3组：对照组常规培养；高碳酸组和缓冲液组于O2－N2－CO2混合气体培养箱中培养，维持培养液PaCO280～100 mmHg；缓冲液组用NaH－CO3缓冲液调节，维持培养液pH值7．2～7．4。分别于给予植物血凝素前和给予植物血凝素细胞培养1、2、4 h（ T0～3）时，随机取4孔，收集细胞和培养液，采用流式细胞仪测定T淋巴细胞坏死率、凋亡率、CD28和CD2阳性率；采用ELISA法测定培养液IL－2、IFN－γ、IL－4和IL－10的浓度。结果3组T细胞T0～3时坏死率和凋亡率比较差异无统计学意义（P＞0．05）。与对照组比较，高碳酸组和缓冲液组T1～3时培养液IL－2和FN－γ的浓度降低，IL－4和IL－10的浓度升高，T2，3时T淋巴细胞CD28和CD2阳性率降低（P＜0．05或P＜0．01）；与高碳酸组比较，缓冲液组T1～3时培养液IL－2和IFN－γ的浓度升高，IL－4和IL－10的浓度降低（ P＜0．05或P＜0．01），各时点T淋巴细胞CD28和CD2阳性率差异无统计学意义（ P＞0．05）。结论高碳酸因素可能通过抑制CD28／B7和CD2／CD48（LFA－3）信号转导通路，减少T淋巴细胞的活化，这是CO2直接作用和其酸性环境共同完成的。Objective To investigate the effects of high carbonate factors on CD 28/B7 and CD2/CD48（LFA-3） signal transduction pathways of activated T lymphocytes in rats .Methods Healthy Wister rats , weighting 200-250 g were sclected .Blood sample was taken from pe-ripheral vein and T lymphocytes was separated in each rat .The T lymphocytes were randomly divided into 3 groups：control group （ group C ） , T cells were conventional cultured;hypercap-nia group （group H）, T cells were exposed to a mixture of O2-N2 and CO2 and the culture media keep PaCO 2 between 80-100 mmHg;and buffering media group （ group pH ） , T cells were incubated as described as group T , and the culture media were also buffered to pH 7.2-7.4 with NaHCO3.In all the three groups, the T cells were incubated for 0 h（T0）, 1 h（T1）, 2 h（T2） or 4 h（T3） at 37 ℃, respectively.After incubation, the T cells and culture media were harvested.The apoptosis and CD28 and CD2 were detected by flow cytometry.The cyto-kines of culture media were examined by ELISA .Results There was no significant difference in death rate and apoptosis rate at T0,1,2,3 among the 3 groups （P〉0.05）.Compared with group C,the media IL-2 and INF-γconcentrations at T 1,2,4 were significantly decreased , the IL-4 and IL-10 concentrations were significantly increased , the expression of CD28and CD2 at T 2,3were significantly decreased in H and pH groups （P〈0.05or P〈0.01 ）.Compared with group H, the media IL-2 and INF-γconcentrations at T1,2,4 were significantly increased, the IL-4 and IL-10 concentrations were significantly decreased （ P〈0.05 or P〈0.01 ） , and there was no significant difference in expression of CD 28 and CD2 at each point in time in pH groups （ P〉0.05 ） .Conclusion The high carbonate factors maybe depress CD 28 and CD2 expression and reduce T lymphocytes activation , buffering the media can reduce the effects of hypercapnia on T lymphocytes .

关 键 词：高碳酸血 T淋巴细胞 抗原 CD28 抗原 CD2 

分 类 号：R655.3[医药卫生—外科学]