晚期氧化蛋白产物抑制CD4＋CD25＋调节性T细胞的增殖  

Advanced oxidation protein products inhibit the proliferation of CD4 ＋ CD25 ＋ regulatory T cells

作　　者：白晓峰[1] 冯梅[1] 李凯[1] 石华[1] 王元林[1] 孙兆林[1] 徐述雄[1] 

机构地区：[1]贵州省人民医院泌尿外科,贵阳550002

出　　处：《中华实验外科杂志》2018年第1期123-126,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金（81260112）;贵州省优秀科技教育人才省长资金项目[黔省专合字（2009）31];贵州省卫生和计划生育委员会基金（gzwjkj-2015-01-023）

摘　　要：目的探讨晚期氧化蛋白产物（AOPP）对CD4＋CD25＋调节性T细胞（Tregs）增殖的影响及机制，为临床上采取措施保持终末期肾病（ESRD）患者体内Tregs数量和功能正常奠定基础。方法从人外周血单个核细胞分离Tregs和CD4＋CD25－ T细胞（作为效应T细胞，Teff）、诱导树突状细胞（DC）后做如下处理：Tregs按5×104个细胞/孔培养于U型底96孔板，加入经丝裂霉素灭活的负载同种异体抗原的DC 5×104个细胞/孔和IL-2 100 U/ml，依据是否加入AOPP分为3组：对照组（加入血清白蛋白200 μg/ml）；AOPP组（加入AOPP 200 μg/ml）、抗氧化组[100 μmol/L还原型烟酰胺腺嘌呤二核苷酸（NADPH）氧化酶抑制剂DPI预处理1 h后再加入200 μg/ml AOPP]。3H-TdR掺入法检测各组Tregs的增殖能力和免疫抑制功能，Western blot检测磷酸化细胞外信号调节激酶1/2（p-ERK1/2）、磷酸化蛋白激酶B（p-Akt）、磷酸化信号转导与转录激活因子5（p-STAT5）和p27kip1等表达情况。 结果在AOPP存在的情况下，AOPP组Tregs 3H-TdR掺入率为（36 213.3±3 385.9）每分钟放射性荧光闪烁计数（cpm），较对照组（65 557.0±3 981.6） cpm低（P＝0.001）。抗氧化组Tregs 3H-TdR掺入率为（44 827.0±3 694.6） cpm，较AOPP组高（P＝0.041），低于对照组（P＝0.003）。AOPP组在Tregs∶Teff为1∶1和1/2∶1的情况下，Tregs对Teff的抑制率分别为（27.3±4.0）%和（23.9±3.7）%，与同比例情况下的对照组抑制率[（68.5±7.9）%和（62.9±5.0）%]比较，差异均有统计学意义（P＝0.001、0.001）。AOPP组Tregs内p-ERK、p-Akt和p-STAT5的表达较对照组低（P＝0.009、0.021、0.036），而p27kip1表达较对照组高（P＝0.036）。抗氧化组Tregs内p-ERK、p-Akt和p-STAT5的表达较AOPP组显著性上调（P＝0.034、0.030、0.042），但仍低于对照组（P＝0.043、0.039、0.007）；而p27kip1表达较AOPP组低（P＝0.048），但高于对照组（P＝0.047）。 �Objective To investigate the effect and mechanism of advanced oxidation protein products （AOPP） on the proliferation of CD4＋ CD25＋ regulatery T cells （Tregs）.MethodsTregs and CD4＋ CD25- T cells （worked as effective T cells, Teff） were isolated, and dendrtic cells （DCs） were induced from the human peripheral blood mononuclear cells. Tregs （5×104/well） were co-cultured with Mitomycin-inactivated DCs loaded with allogeneic antigen and IL-2 （100 U/ml）, and according to the presence of AOPP into 3 different groups： Control group （200 μg/ml serum albumin）, AOPP group （200 μg/ml AOPP）, Antioxidant group [pretreated with 100 μmol/L diphenyleneiodonium （DPI） 1 h before AOPP was added]. The incorporation of [3H]thymidine was used to evaluate the proliferation of Tregs and the suppressive function of Tregs on the Teff. Western blotting was used to observe the expression of phosphorylated extracellular signal-regulated kinase 1/2 （p-ERK1/2）, phosphorylated protein kinase B （p-Akt）, phosphorylated signal transducer and activators of transcription 5 （p-STAT5） and p27kip1.ResultsIn AOPP group, the incorporation of [3H]thymidine was （36213.3±3 385.9） cpm, which was significantly lower than that [（65 557.0±3 981.6） cpm] of Control group （P=0.001）. In Antioxidant group, the value was （44 827.0±3 694.6） cpm, which was significantly higher than that of AOPP group （P=0.041）, but lower than that of Control group （P=0.003）. When Tregs and Teff were incubated at the ratio of 1∶1 and 1/2∶1, the suppressive ratios of Tregs on the Teff in AOPP group were （27.3±4.0）% and （23.9±3.7）%, which were significantly lower than those [（68.5±7.9）% and （62.9±5.0）%] of Control group （P=0.001, 0.001）. Compared with Control group, addition of AOPP significantly downregulated the expression of p-ERK, p-Akt, p-STAT5 and upregulated the expression of p27kip1 in Tregs （P=0.009, 0.021, 0.036, 0.036）. After antioxidants was added, the expressi

关 键 词：晚期氧化蛋白产物 调节性T细胞 增殖 

分 类 号：R692[医药卫生—泌尿科学]