CAJ | 학술논문

目的探讨干扰素(IFN-γ)联合白消安诱导建立的重型再生障碍性贫血(AA)模型小鼠脾脏中CD4+CD25+调节性T(Treg)细胞的凋亡情况以及可能的机制.方法应用IFN-γ腹腔注射联合白消安灌胃诱导建立BALB/c重型AA模型小鼠(联合组),以正常组、IFN-γ组和白消安组(各16只)为对照.采用免疫磁珠法分选各组小鼠脾脏Treg细胞,应用流式细胞术检测各组小鼠脾脏Treg细胞的分选纯度及凋亡率,通过Western blot法检测各组小鼠脾脏Treg细胞的蛋白激酶B(Akt)和转化生长因子-β(TGF-β)的表达水平.结果联合组小鼠脾脏Treg细胞的凋亡率为(33.21±0.65)％,较正常组[(27.38±1.89)％]、IFN-γ组[(17.62±1.05)％]和白消安组[(21.58±0.64)％]升高,差异有统计学意义(P＜0.05),而Akt、TGF-β蛋白表达水平分别是0.30±0.05和0.17±0.05,较其他3组降低,差异有统计学意义(P＜0.05).结论 IFN-γ联合白消安诱导的重型AA模型小鼠的脾脏Treg细胞存在过度凋亡现象,这可能是AA患者Treg细胞减少的原因之一；而Akt和TGF-β的表达水平下调可能在其过度凋亡中起一定作用,相关实验结果可能为AA免疫治疗提供新的理论思路.
목적 탐토간우소(IFN-γ)연합백소안유도건립적중형재생장애성빈혈(AA)모형소서비장중CD4+CD25+조절성T(Treg)세포적조망정황이급가능적궤제.방법 응용IFN-γ복강주사연합백소안관위유도건립BALB/c중형AA모형소서(연합조),이정상조、IFN-γ조화백소안조(각16지)위대조.채용면역자주법분선각조소서비장Treg세포,응용류식세포술검측각조소서비장Treg세포적분선순도급조망솔,통과Western blot법검측각조소서비장Treg세포적단백격매B(Akt)화전화생장인자-β(TGF-β)적표체수평.결과 연합조소서비장Treg세포적조망솔위(33.21±0.65)％,교정상조[(27.38±1.89)％]、IFN-γ조[(17.62±1.05)％]화백소안조[(21.58±0.64)％]승고,차이유통계학의의(P＜0.05),이Akt、TGF-β단백표체수평분별시0.30±0.05화0.17±0.05,교기타3조강저,차이유통계학의의(P＜0.05).결론 IFN-γ연합백소안유도적중형AA모형소서적비장Treg세포존재과도조망현상,저가능시AA환자Treg세포감소적원인지일；이Akt화TGF-β적표체수평하조가능재기과도조망중기일정작용,상관실험결과가능위AA면역치료제공신적이론사로.
Objective To explore the apoptosis and its mechanisms of spleen CD4 + CD25 + regulatory T (Treg) cells in severe aplastic anemia (SAA) mouse model induced by interferon (IFN-γ) in combination with busulphan (BU).Methods The BALB/c female mice SAA model was induced by intraperitoneal injection with IFN-γ and intragastric administration with BU (combined group,n=1 6),with BU group (n=16),IFN-γ group (n=16) and normal group (n=16) as controls.Spleen Treg cells were purified by using of immunomagnetic beads.Apoptosis was detected by flow cytometry.Akt and TGF-β expression was measured by Western blot.Results Apoptosis of spleen Treg cells in combined group [(33.21±0.65)％]was significantly higher than that in BU group [(21.58±0.64)％],IFN-γ group [(17.62± 1.05)％],and control [(27.38± 1.89)％] (P ＜ 0.05).A significantly lower expression of Akt and TGF-β protein was also seen in combined group (0.30±0.05 and 0.17±0.05),as compared to the other three groups (P ＜ 0.05).Conclusion Excessive apoptosis of Treg cells was found in SAA mouse model,which may be a cause of Treg cells decrease in patients with AA.The down-regulated expression of Akt and TGF-β could play a role in increased apoptosis of Treg cells.Our data may provide a new treatment strategy in AA.