左归丸对老年大鼠CD4^+T细胞IL-2基因启动子区域甲基化的影响  

Effect of ZuoGui Pill on the regulation to the specific CpG sites methylation in interleukin-2 gene promoter region of aged rats CD4^+ Tcells

作　　者：龚张斌[1] 徐品初[2] 韩志芬 金国琴 

机构地区：[1]上海中医药大学基础医学院生化教研室,上海201203 [2]上海中医药大学附属曙光医院

出　　处：《中国老年学杂志》2015年第3期710-712,共3页Chinese Journal of Gerontology

基　　金：国家自然科学基金(No.81001565;30873317);上海市教育委员会重点学科建设项目(No.J50301);"085"一流学科建设科技创新支撑计划(No.085ZY1204);上海中医药大学教师学术共同体建设项目(No.2013JXG02)

摘　　要：目的研究左归丸对自然衰老大鼠CD4+T细胞白细胞介素(IL)-2基因启动子Cp G位点甲基化水平的影响。方法以自然衰老大鼠为模型,分为青年对照组、老年对照组、老年左归组。流式细胞仪分选各组大鼠CD4+T细胞,以抗CD3联合抗CD28进行刺激。Q Real time RTPCR法检测IL-2 mRNA的表达;ELISA法检测DNA甲基转移酶(DNMTs)活性,甲基化特异性PCR(MSP)法检测IL-2基因启动子Cp G位点甲基化水平。结果抗CD3联合抗CD28刺激24 h后,与青年对照组比较,老年对照组CD4+T细胞IL-2 mRNA表达显著降低(P<0.01);DNMTs活性显著增强(P<0.01),IL-2基因SET1区域甲基化水平显著升高(P<0.01)。与老年对照组比较,老年左归组CD4+T细胞IL-2 mRNA表达明显升高(P<0.05);DNMTs活性明显减弱(P<0.05),IL-2基因SET1区域甲基化水平明显降低(P<0.05)。结论左归丸能通过抑制DNMTs活性,降低IL-2基因SET1区域甲基化水平,促进IL-2表达。Objective To investigate whether Zuo Gui Pill( ZGP) elevate interleukin( IL)-2 expression through regulating the Cp G methylation status in the CD4+T cells of aged rat. Methods The IL-2 expressions in CD4+T cells from different aged rats were analyzed,the activity of DNMTs,the Cp G methylation status in the different site of IL-2 promoter region with MSP were measured. Results The expression of IL-2 mRNA of aged cells was lower than that of young CD4+T cells( P < 0. 01). Methylation ratio of specific Cp G sites(- 254,- 118,- 47,- 44,+ 58) within the IL-2 promoter region were significantly higher in the old rats compared to those of the young ones. After treated with ZGP,the methylation ratio of specific Cp G sites were significantly decreased compared to those of the old rats. Conclusions DNA hyper-methylation status on the specific Cp G sites could play an important role in the cellular immune function with aging. ZGP could elevate the IL-2 transcription level through inhibiting the DNMTs activity,regulating the methylation modification of the specific Cp G sites.

关 键 词：衰老 白细胞介素2 表观遗传 DNA甲基化 

分 类 号：R285.5[医药卫生—中药学]