CAJ | 학술논문

背景：SIRT6/NF-κB信号轴是细胞衰老的调控通路，但其在造血干/祖细胞(hematopoietic stem and progenitor cel ，HSC/HPC)衰老中的作用尚不清楚。目的：探讨SIRT6/NF-κB信号通路在三丁基过氧化氢体外诱导造血干/祖细胞衰老中的作用。方法：免疫磁性分选法分离、纯化小鼠Sca-1+ HSC/HPC。用100μmol/L三丁基过氧化氢体外诱导Sca-1+HSC/HPC构建体外衰老模型，通过衰老相关β-半乳糖苷酶细胞化学染色、流式细胞术分析细胞周期、造血祖细胞混合集落培养确定三丁基过氧化氢诱导 Sca-1+HSC/HPC 衰老的生物学作用。荧光定量 PCR 及Western blotting检测衰老调控分子SIRT6、NF-κB mRNA及蛋白的表达。结果与结论：与对照组比较，衰老组Sca-1+ HSC/HPC β-半乳糖苷酶染色阳性细胞数、G0/G1期细胞比例增高，形成造血祖细胞混合集落数量减少；SIRT6 mRNA及蛋白表达下调，NF-κB mRNA及蛋白表达上调。结果表明三丁基过氧化氢可能通过调控SIRT6/NF-κB信号通路发挥诱导Sca-1+HSC/HPC衰老作用。
배경：SIRT6/NF-κB신호축시세포쇠로적조공통로，단기재조혈간/조세포(hematopoietic stem and progenitor cel ，HSC/HPC)쇠로중적작용상불청초。목적：탐토SIRT6/NF-κB신호통로재삼정기과양화경체외유도조혈간/조세포쇠로중적작용。방법：면역자성분선법분리、순화소서Sca-1+ HSC/HPC。용100μmol/L삼정기과양화경체외유도Sca-1+HSC/HPC구건체외쇠로모형，통과쇠로상관β-반유당감매세포화학염색、류식세포술분석세포주기、조혈조세포혼합집락배양학정삼정기과양화경유도 Sca-1+HSC/HPC 쇠로적생물학작용。형광정량 PCR 급Western blotting검측쇠로조공분자SIRT6、NF-κB mRNA급단백적표체。결과여결론：여대조조비교，쇠로조Sca-1+ HSC/HPC β-반유당감매염색양성세포수、G0/G1기세포비례증고，형성조혈조세포혼합집락수량감소；SIRT6 mRNA급단백표체하조，NF-κB mRNA급단백표체상조。결과표명삼정기과양화경가능통과조공SIRT6/NF-κB신호통로발휘유도Sca-1+HSC/HPC쇠로작용。
BACKGROUND:SIRT6/NF-κB is the important signal axis to cellsenescence, but the effect of SIRT6/NF-κB signal axis to hematopoietic stem and progenitor cell(HSC/HPC) senescence is unclear. OBJECTIVE:To induce (t-BHP) in vitro and to investigate the role of SIRT6/NF-κB signal axis in Sca-1+HSC/HPC senescence induced by tert-butylhydroperoxide in vitro. METHODS:Sca-1+HSC/HPC was isolated and purified by magnetic activated cellsorting. Sca-1+HSC/HPC senescence was induced by 100μmol/L tert-butylhydroperoxide in vitro. The senescence-associatedβ-Galactosidase staining, cellcycle analysis and culture of mixed hematopoietic progenitor cellwere used to investigate the biological effects of tert-butylhydroperoxide on Sca-1+HSC/HPC senescence. The expression of senescence associated SIRT6, NF-κB mRNA and protein was examined by real-time fluorescence quantitative PCR and western blot assay. RESULTS AND CONCLUSION:Compared with control group, the percentage of positive cells expressing SA-β-Gal and cells in G0/G1 phase increased and the number of forming colony of mixed hematopoietic progenitor decreased in the aging group. It showed lower expression of SIRT6 and higher expression of NF-κB in the aging group. The SIRT6/NF-κB signal axis may play a key role in the Sca-1+ HSC /HPC senescence inducted by tert-butylhydroperoxide.