人肝再生增强因子对人外周血单个核细胞增殖的影响及其机制  

Effect of Human Augmenter of Liver Regeneration on Proliferation of Human PBMCs and Its Mechanism

作　　者：王新国[1] 刘杞[1] 孙航[1] 梁珊[1] 谢松丽[1] 张宏华[1] 

机构地区：[1]重庆医科大学病毒性肝炎研究所教育部感染性疾病分子生物学重点实验室,重庆400010

出　　处：《中国生物制品学杂志》2009年第10期961-963,共3页Chinese Journal of Biologicals

基　　金：国家自然科学基金(30570826);重庆市自然科学基金(CSTC2008BB5228)

摘　　要：目的探讨人肝再生增强因子(hALR)对人外周血单个核细胞(PBMC)增殖的影响及其机制。方法梯度离心分离PBMC,将细胞分为正常对照组、刀豆蛋白A(ConA)(5mg/L)组和ConA(5mg/L)+hALR(30mg/L)组,分别培养10min、30min、1h、2h、4h后,MTT法检测各组各时间点细胞增殖水平,钙荧光指示剂法检测细胞内游离Ca2+浓度的变化,Western blot法检测细胞内胞外信号调节激酶(ERK)的磷酸化程度。结果4h内,各组细胞的增殖水平差异无统计学意义,但ConA组细胞增殖水平已表现出逐渐升高的趋势。正常对照组细胞内Ca2+浓度在加入Fluo-3/AM后2h达高峰,ConA组1h达高峰,ConA+hALR组10min达高峰。正常对照组细胞内磷酸化的ERK随培养时间的延长而逐渐减少;ConA组细胞内磷酸化的ERK在30min达高峰,之后逐渐降低;ConA+hALR组细胞内磷酸化的ERK在2h前明显低于ConA组。结论hALR可能通过影响细胞内Ca2+浓度的变化峰值和磷酸化的ERK,来抑制人PBMC的免疫功能,从而影响其增殖。Objective To investigate the effect of human augmenter of liver regeneration（hALR）on proliferation of human PBMCs and its mechanism.Methods The PBMCs of healthy volunteers were separated by gradient centrifugation and divided into normal control,ConA（5mg/L）and ConA（5mg /L）＋ hALR（30 mg /L）groups.The proliferation levels of PBMCs 10 min,30 min,1 h,2 h and 4 h after culture were determined by MTT method,the free calcium ion concentration in PBMCs by calcium fluorescent indicator,and the phosphorylation level of extracellular signal regulatory kinase（ERK）in PBMCs by Western blot.Results No significant differences were observed in the proliferation levels of PBMCs in various groups within 4 h after culture,though those in ConA group showed an increasing tendency.The calcium ion concentration in PBMCs in normal control group reached a peak value 2 h,while those in ConA and ConA ＋ hALR groups reached peak values 1 h and 10 min after addition with Fluo-3 /AM respectively.The phosphorylation level of ERK in PBMCs in normal control group decreased gradually with the increasing time for culture,while that in ConA group reached a peak value 30 min after culture then decreased gradually.However,the phosphorylation level of ERK in PBMCs in ConA ＋ hALR group within 2 h after culture was significantly lower than that in ConA group.Conclusion hALR may inhibit the immune function and proliferation of human PBMCs by influencing the peak value of calcium ion concentration and phosphorylation level of ERK in the cells.

关 键 词：人肝再生增强因子 外周血单个核细胞 细胞增殖 钙离子 胞外信号调节激酶 

分 类 号：R392.32[医药卫生—免疫学]