【摘要】：目的探討氧化低密度脂蛋白(ox-LDL)體外誘導(dǎo)造血干細胞(HSCs)衰老的可能機制。方法用免疫磁性分選法分離純化小鼠HSC,與ox-LDL共培養(yǎng),采用β-半乳糖苷酶(SA-β-Gal)染色檢測衰老HSC,流式細胞術(shù)檢測HSC細胞周期分布,混合集落培養(yǎng)(CFU-Mix)檢測HSC混合集落形成能力。流式細胞術(shù)和免疫熒光檢測HSC產(chǎn)生活性氧(ROS)的量,酶學(xué)比色法檢測HSC培養(yǎng)上清液超氧化物歧化酶(SOD)、谷胱甘肽過氧化物酶(GSH-Px)及丙二醛(MDA)含量。Southern blot和TRAP-PCR法檢測HSC端粒長度和端粒酶活性。結(jié)果 ox-LDL誘導(dǎo)HSC呈現(xiàn)典型的衰老生物學(xué)表現(xiàn):SA-β-Gal染色陽性細胞率顯著增高(P0.01);G0/G1期比例明顯增加,S期顯著減少(P0.01);CFU-Mix數(shù)量顯著減少(P0.01)。衰老HSC端�？s短(P0.05),端粒酶活性降低(P0.05)。衰老HSC ROS含量顯著增加(P0.01),細胞培養(yǎng)上清液中SOD、GSH-Px活力下降、MDA含量增加(P0.05)。結(jié)論 ox-LDL能通過氧化應(yīng)激誘導(dǎo)HSC衰老,其機制可能與ROS的蓄積及抗氧化酶活性受抑引起端粒功能異常有關(guān)。
[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.
【作者單位】： 重慶醫(yī)科大學(xué)干細胞與組織工程研究室組織學(xué)與胚胎學(xué)教研室;遵義醫(yī)學(xué)院;遵義醫(yī)學(xué)院附屬醫(yī)院腫瘤科;山東省棗莊市臺兒莊區(qū)中醫(yī)院急診科;
【基金】：國家自然科學(xué)基金(81173398,30970872)
【分類號】：R363

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相關(guān)期刊論文 前2條

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