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目的探讨去乙酰化酶SIRT6/核转录因子-κB(NF-κB)信号通路在人参皂苷Rg_1抗辐射致造血干/祖细胞(Sca-1+HSC/HPC)衰老中的作用。方法 C57BL/6小鼠随机分为对照组、模型组和人参皂苷Rg_1组。给予模型组和人参皂苷Rg_1组小鼠全身一次性6.5 Gy ~(60)Coγ线照射,人参皂苷Rg_1组于照射前ip人参皂苷Rg_1 20 mg/kg,每天1次,连续给药7 d,照射后继续ip等剂量人参皂苷Rg_1 7 d。给药结束后第2天,外周血血象指标检测确定人参皂苷Rg_1促进造血恢复情况;免疫磁性分选法分离纯化各组Sca-1+HSC/HPC,造血祖细胞混合性集落(CFU-Mix)培养、细胞周期分析和衰老相关β-半乳糖苷酶(SA-β-Gal)染色分析人参皂苷Rg_1抗辐射损伤致Sca-1+HSC/HPC衰老的生物学作用;荧光定量PCR及Western blotting法检测衰老调控分子SIRT6、NF-κB mRNA及蛋白的表达。结果辐射后,与对照组相比,模型组小鼠外周血象恢复缓慢,Sca-1+HSC/HPC出现细胞衰老特征,G0/G1期细胞比例及SA-β-Gal染色阳性率增高,CFU-Mix数量下降,SIRT6 mRNA及蛋白表达下调,NF-κB mRNA及蛋白表达上调。与模型组比较,人参皂苷Rg_1组小鼠白细胞(WBC)、红细胞(RBC)、血小板(PLT)数量增高,Sca-1+HSC/HPC G_0/G_1期细胞比例及SA-β-Gal染色阳性率下降,CFU-Mix数量升高,SIRT6 mRNA及蛋白表达上调,NF-κB mRNA及蛋白表达下调。结论人参皂苷Rg_1可通过调控SIRT6/NF-κB信号通路发挥抗辐射致Sca-1+HSC/HPC衰老的作用。
Objective To investigate the role of SIRT6 / NF-κB signaling pathway in the senescence of hematopoietic stem / progenitor cells (Sca-1 + HSC / HPC) induced by ginsenoside Rg 1. Methods C57BL / 6 mice were randomly divided into control group, model group and ginsenoside Rg_1 group. The mice in the model group and the ginsenoside Rg_1 group were given a one-time whole body radiation of 6.5 Gy ~ (60) Co γ-rays. The ginsenoside Rg_1 group was given ip ginsenoside Rg_1 20 mg / kg once a day for 7 days after irradiation After continued ip ip and other doses of ginsenoside Rg_1 7 d. On the second day after the administration, blood gindex of peripheral blood was detected to determine the recovery of hematopoietic cells by ginsenoside Rg_1. Immunomagnetic separation was used to separate and purify the Sca-1 + HSC / HPC and hematopoietic progenitor cell mixed colonies (CFU-Mix) The biological effects of ginsenoside Rg 1 on the senescence of Sca-1 + HSC / HPC induced by radiation were analyzed by cell cycle analysis and aging-related β-galactosidase (SA-β-Gal) staining. The expressions of SIRT6, NF-κB mRNA and protein were detected by immunohistochemistry. Results Compared with the control group, the peripheral blood of mice in the model group recovered slowly. The cell senescence characteristics of Sca-1 + HSC / HPC were observed. The percentage of cells in G0 / G1 phase and the positive rate of SA-β-Gal staining were increased. The number of Mix decreased, the expression of SIRT6 mRNA and protein was down-regulated, and the expression of NF-κB mRNA and protein was up-regulated. Compared with model group, the number of white blood cells (WBC), red blood cells (RBC) and platelets (PLT) of mice in ginsenoside Rg_1 group increased, the percentage of cells in Sca-1 + HSC / HPC G0 / Decreased, the number of CFU-Mix increased, SIRT6 mRNA and protein expression increased, NF-κB mRNA and protein expression down. Conclusion Ginsenoside Rg_1 can play an important role in the anti-aging of Sca-1 + HSC / HPC by regulating SIRT6 / NF-κB signaling pathway.