目的:研究不同浓度D-核糖对K562细胞增殖活力、细胞内ATP水平及线粒体呼吸链相关基因表达的影响。方法:使用不同浓度D-核糖和D-葡萄糖分别在基础培养基或PBS中孵育K562细胞6、24或48h,利用CCK-8、ATP检测试剂盒及实时荧光定量PCR(RT-qPCR)分别检测K562细胞的增殖活力、细胞内ATP水平及线粒体呼吸链基因(MTND6、MTATP8)的表达。结果:与D-葡萄糖促进增殖作用不同,低浓度D-核糖(10-25mmol/L)在24h内对细胞增殖无显著影响,但48h后呈明显抑制作用。ATP检测结果显示,基础培养基中孵育24h,与葡萄糖组普遍增加ATP水平不同,仅50mmol/LD-核糖组ATP水平增高;48h后虽然对照组ATP几乎耗尽,但10mmol/L和20mmol/L核糖组中ATP明显高于对照组。在无糖(PBS)条件下,与D-葡萄糖显著提升ATP水平不同,D-核糖无法有效维持ATP水平。RT-qPCR显示,D-核糖显著抑制线粒体呼吸链基因MTATP8的表达(P＜0.01),但对MTND6表达无影响。结论:高浓度D-核糖持续作用可抑制K562细胞增殖及呼吸链基因表达,对促进ATP生成贡献不大,但可能会降低ATP的利用。该研究为进一步深入理解D-核糖在维持代谢稳态中的功能奠定实验基础。
Objective: To study the effects of different concentrations of D-ribose on the proliferative viability, intracellular ATP level and mitochondrial respiratory chain-related gene expression of K562 cells. Methods: K562 cells were incubated with different concentrations of D-ribose and D-glucose in basal medium or PBS for 6, 24 or 48 hours, and the proliferation viability, intracellular ATP level and the expression of genes related to mitochondrial respiratory chain (MTND6, MTATP8) were detected by using CCK-8, ATP detection kit and real-time fluorescence quantitative PCR (RT-qPCR), respectively. Results: Unlike the proliferative effect induced by D-glucose, low concentrations of D-ribose (10-25 mmol/L) had no significant effect on cell proliferation within 24 hours, but significantly inhibited cell proliferation after 48 hours. The results of ATP detection showed that the ATP level was generally increased in the basal medium for 24 h, which was different from that in the glucose group, and only the ATP level was increased in the 50 mmol/L D-ribose group; after 48 h, although the ATP in the control group was almost depleted, the ATP in the 10 mmol/L and 20 mmol/L ribose groups was significantly higher than that in the control group. Under the condition of sugar-free (PBS), unlike D-glucose, which significantly increased ATP levels, D-ribose could not effectively maintain ATP levels. RT-QPCR showed that D-ribose significantly inhibited the expression of mitochondrial respiratory chain gene MTATP8 (P＜0.01), but had no effect on the expression of MTND6. Conclusion: High concentration of D-ribose can inhibit the proliferation of K562 cells and the expression of respiratory chain genes, and has little contribution to the promotion of ATP production, but may reduce the utilization of ATP. This study lays an experimental foundation for further understanding the function of D-ribose in maintaining metabolic homeostasis.
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