摘要
目的观察两种共刺激分子CD137及CD28对D-半乳糖致亚急性衰老模型小鼠及自然衰老小鼠T细胞活化后的bcl-2表达的调控,分析这两种共刺激分子调节衰老T细胞凋亡的可能机制。方法7周龄BALB/c雄性小鼠每日背部皮下注射D-半乳糖溶液(120mg/kg,溶于0.1ml蒸馏水中),连续注射5个月,建立亚急性衰老模型组;自然衰老组为16月龄BALB/c雄性小鼠。分离小鼠的脾脏T细胞,分别用conA+IgG、conA+CD137单抗、conA+CD28单抗体外活化,48h后用流式细胞术及半定量RT-PCR分析各组T细胞bcl-2的表达。结果(1)衰老模型组T细胞经conA+IgG、conA+CD137单抗、conA+CD28单抗活化48h后,胞内bcl-2蛋白表达率分别为(24.4±1.5)%、(34.4±2.4)%、(45.1±2.7)%,自然衰老组T细胞经conA+IgG、conA+CD137单抗、conA+CD28单抗活化48h后,胞内bcl-2蛋白表达率分别(19.6±2.0)%、(26.3±1.9)%、(48.5±2.2)%;(2)衰老模型组T细胞经conA+IgG、conA+CD137单抗、conA+CD28单抗活化48h后,胞内bcl-2mRNA表达率(IOD(bcl-2)/IOD(β-actin)分别为0.309±0.039、0.547±0.036、0.780±0.041,自然衰老组T细胞经conA+IgG、conA+CD137单抗、conA+CD28单抗活化48h后,胞内bcl-2mRNA表达率分别0.283±0.038、0.535±0.041、0.771±0.063。结论CD137单抗及CD28单抗,均可以显著提高衰老模型组及自然衰老组小鼠T细胞bcl-2mRNAbcl-2蛋白的表达,这可能是CD137分子及CD28分子抑制衰老T细胞凋亡、维持衰老T细胞功能的主要机制之一。
Objective To observe the effect of the costimulatory molecules CD137 and CD28 on Bcl-2 expression in T cells of mice with D-galactose-induced aging and natural aging. Methods Seven-week-old male BALB/c mice were injected daily with D-galactose （120 mg/kg） for 5 months to induce subacute aging, and 16-month-old male BALB/c mice served as the natural aging group. The spleen T cells of were isolated from the rats and stimulated in vitro with conA ＋IgG, conA＋CD 137 mAb or conA＋CD28 rnA, b for 48 h. Bcl-2 expression of T cells was detected by flow cytometry and semi-quantitative RT-PCR. Results In the subacute aging group, Bcl-2 protein expression rate in the T cells stimulated with conA＋IgG, conA＋CD 137 mAb and conA＋CD28 mAb were （24.4 ± 1.5）%, （34.4 ±2.4）%, and （45.1 ±2.7）%, which were （19.6 ±2.0）%, （26.3±1.9）%, and （48.5±2.2）% in the natural aging group, respectively. In vitro stimulation with conA＋IgG, conA＋CD137 mAb and conA＋CD28 mAb resulted in expression rate ofbcl-2 mRNA in the T cells of 0.309±0.039, 0.547±0.036, and 0.780±0.041 in the subacute aging group, and 0.283±0.038, 0.535±0.041, and 0.771±0.063 in the natural aging group, respectively. Conclusion CD 137 and CD28 can promote bcl-2 expression at both mRNA and protein level in T cells in aging mice, which may be one of the mechanisms through which CD137 and CD28 molecules inhibit aging T cell apoptosis and maintain their function.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2007年第9期1427-1430,共4页
Journal of Southern Medical University
