CAJ | 학술논문

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CpG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 RT－PCR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CpG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。
목적：연구좌귀환대자연쇠로대서CD4＋T세포백세포개소（ IL）－2기인계동자CpG위점갑기화수평적영향。방법이자연쇠로대서위모형，분위청년대조조、노년대조조、노년좌귀조。류식세포의분선각조대서 CD4＋T세포，이항 CD3연합항 CD28진행자격。 Q Real time RT－PCR법검측IL－2 mRNA적표체；ELISA법검측DNA갑기전이매（ DNMTs）활성，갑기화특이성PCR（ MSP）법검측IL－2기인계동자CpG위점갑기화수평。결과항CD3연합항CD28자격24 h후，여청년대조조비교，노년대조조CD4＋T세포IL－2 mRNA 표체현저강저（P＜0．01）；DNMTs활성현저증강（P＜0．01），IL－2기인SET1구역갑기화수평현저승고（P＜0．01）。여노년대조조비교，노년좌귀조 CD4＋T 세포 IL－2 mRNA 표체명현승고（P＜0．05）；DNMTs활성명현감약（P＜0．05），IL－2기인SET1구역갑기화수평명현강저（P＜0．05）。결론좌귀환능통과억제DNMTs활성，강저IL－2기인SET1구역갑기화수평，촉진IL－2표체。
Objective To investigate whether ZuoGui Pill ( ZGP) elevate interleukin( IL)-2 expression through regulating the CpG 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 CpG methylation status in the different site of IL-2 promoter region with MSP were measured.Results The expres-sion of IL-2 mRNA of aged cells was lower than that of young CD4+T cells (P<0.01).Methylation ratio of specific CpG 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 CpG sites were significantly decreased compared to those of the old rats.Conclusions DNA hy-per-methylation status on the specific CpG 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 CpG sites.