论文部分内容阅读
目的探讨人参皂苷Rg1(Rg1)诱导人白血病K562细胞复制性衰老特征性变化。方法四甲基偶氮唑盐(MTT)法筛选Rg1对K562细胞增殖抑制的最佳浓度及时间,以此浓度和作用时间诱导K562细胞衰老。流式细胞术检测Rg1对细胞增殖周期的影响;衰老相关β-半乳糖苷酶(SA-β-Gal)染色检测阳性细胞百分率;Southern blotting检测端粒长度;Western blotting检测复制性衰老信号通路相关P21、P53与Rb蛋白表达;透射电镜观察人白血病K562细胞衰老超微形态学改变。结果 Rg1在体外能明显抑制K562细胞增殖,其最佳作用浓度为20μmol/L,时间为48h;诱导后的K562细胞阻滞于G2/M期;SA-β-Gal染色阳性细胞百分率增加(P<0.05);复制性衰老通路相关蛋白P21、P53和Rb表达上调(P<0.05);端粒长度缩短加速(P<0.05);经诱导细胞呈现胞体增大,溶酶体体积增大、数目增多,线粒体体积增大等衰老形态学变化。结论 Rg1可能经由p53-p21-Rb信号通路诱导K562细胞呈现复制性衰老特征。
Objective To investigate the characteristic changes of reproductive senescence induced by ginsenoside Rg1 (Rg1) in human leukemia K562 cells. Methods MTT method was used to screen the optimal concentration and time of Rg1 to inhibit the proliferation of K562 cells. The K562 cells were induced to senescence at this concentration and time. Flow cytometry was used to detect the effect of Rg1 on the cell cycle. The percentage of positive cells was detected by aging-related β-galactosidase (SA-β-Gal) staining. The telomere length was detected by Southern blotting and Western blotting P21, P53 and Rb protein expression; Transmission electron microscopy observation of human leukemia K562 cells aging ultrastructural changes. Results Rg1 could inhibit the proliferation of K562 cells in vitro. The optimal concentration of Rg1 was 20μmol / L for 48h. The induced K562 cells were arrested in G2 / M phase. The percentage of P-gp positive cells increased (P <0.05). The expressions of P21, P53 and Rb were up-regulated (P <0.05), the telomere length was shortened and accelerated (P <0.05) Increased, mitochondrial volume and other morphological changes of aging. Conclusion Rg1 may induce the replication-like senescence of K562 cells via the p53-p21-Rb signaling pathway.