氧化低密度脂蛋白通过氧化应激诱导小鼠造血干细胞衰老
发布时间：2018-11-22 08:23
【摘要】：目的探讨氧化低密度脂蛋白(ox-LDL)体外诱导造血干细胞(HSCs)衰老的可能机制。方法用免疫磁性分选法分离纯化小鼠HSC,与ox-LDL共培养,采用β-半乳糖苷酶(SA-β-Gal)染色检测衰老HSC,流式细胞术检测HSC细胞周期分布,混合集落培养(CFU-Mix)检测HSC混合集落形成能力。流式细胞术和免疫荧光检测HSC产生活性氧(ROS)的量,酶学比色法检测HSC培养上清液超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)及丙二醛(MDA)含量。Southern blot和TRAP-PCR法检测HSC端粒长度和端粒酶活性。结果 ox-LDL诱导HSC呈现典型的衰老生物学表现:SA-β-Gal染色阳性细胞率显著增高(P0.01);G0/G1期比例明显增加,S期显著减少(P0.01);CFU-Mix数量显著减少(P0.01)。衰老HSC端粒缩短(P0.05),端粒酶活性降低(P0.05)。衰老HSC ROS含量显著增加(P0.01),细胞培养上清液中SOD、GSH-Px活力下降、MDA含量增加(P0.05)。结论 ox-LDL能通过氧化应激诱导HSC衰老,其机制可能与ROS的蓄积及抗氧化酶活性受抑引起端粒功能异常有关。
[Abstract]:Objective to investigate the possible mechanism of (HSCs) senescence induced by oxidized low density lipoprotein (ox-LDL). Methods HSC, and ox-LDL were isolated and purified by immunomagnetic sorting method, and the distribution of HSC cell cycle was detected by SA- 尾 Gal staining. Mixed colony culture (CFU-Mix) was used to detect the ability of HSC mixed colony formation. Flow cytometry and immunofluorescence were used to detect the amount of reactive oxygen (ROS) produced by HSC, and the superoxide dismutase (SOD), in superoxide dismutase (SOD),) supernatant of HSC culture was detected by enzymatic colorimetry. Glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) (MDA) content. Southern blot and TRAP-PCR were used to detect the telomere length and telomerase activity of HSC. Results ox-LDL induced HSC showed typical senescence biological manifestations: SA- 尾-Gal staining positive cells increased significantly (P0.01), G0/G1 phase increased significantly, S phase decreased significantly (P0.01). The number of CFU-Mix decreased significantly (P0.01). Aging HSC telomere shortening (P0.05), telomerase activity decreased (P0.05). The content of senescent HSC ROS increased significantly (P0.01), the activity of SOD,GSH-Px in supernatant of cell culture decreased, and the content of MDA increased (P0.05). Conclusion ox-LDL can induce HSC senescence through oxidative stress. The mechanism may be related to the abnormal telomere function caused by the accumulation of ROS and the inhibition of antioxidant enzyme activity.
【作者单位】： 重庆医科大学干细胞与组织工程研究室组织学与胚胎学教研室;遵义医学院;遵义医学院附属医院肿瘤科;山东省枣庄市台儿庄区中医院急诊科;
【基金】：国家自然科学基金(81173398,30970872)
【分类号】：R363
[Abstract]:Objective to investigate the possible mechanism of (HSCs) senescence induced by oxidized low density lipoprotein (ox-LDL). Methods HSC, and ox-LDL were isolated and purified by immunomagnetic sorting method, and the distribution of HSC cell cycle was detected by SA- 尾 Gal staining. Mixed colony culture (CFU-Mix) was used to detect the ability of HSC mixed colony formation. Flow cytometry and immunofluorescence were used to detect the amount of reactive oxygen (ROS) produced by HSC, and the superoxide dismutase (SOD), in superoxide dismutase (SOD),) supernatant of HSC culture was detected by enzymatic colorimetry. Glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) (MDA) content. Southern blot and TRAP-PCR were used to detect the telomere length and telomerase activity of HSC. Results ox-LDL induced HSC showed typical senescence biological manifestations: SA- 尾-Gal staining positive cells increased significantly (P0.01), G0/G1 phase increased significantly, S phase decreased significantly (P0.01). The number of CFU-Mix decreased significantly (P0.01). Aging HSC telomere shortening (P0.05), telomerase activity decreased (P0.05). The content of senescent HSC ROS increased significantly (P0.01), the activity of SOD,GSH-Px in supernatant of cell culture decreased, and the content of MDA increased (P0.05). Conclusion ox-LDL can induce HSC senescence through oxidative stress. The mechanism may be related to the abnormal telomere function caused by the accumulation of ROS and the inhibition of antioxidant enzyme activity.
【作者单位】： 重庆医科大学干细胞与组织工程研究室组织学与胚胎学教研室;遵义医学院;遵义医学院附属医院肿瘤科;山东省枣庄市台儿庄区中医院急诊科;
【基金】：国家自然科学基金(81173398,30970872)
【分类号】：R363
【参考文献】
相关期刊论文 前2条
1 陈小莉;李雅慧;王成;李绕明;苏峰;杨庆;贾凤歧;卫立辛;;同型半胱氨酸促进牛主动脉内皮细胞衰老的研究[J];第二军医大学学报;2007年02期
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