本研究旨在探讨二氢杨梅素对骨肉瘤细胞增殖的抑制作用及其机制。采用MTT法检测不同浓度的二氢杨梅素溶液对骨肉瘤细胞增殖的抑制作用；采用倒置显微镜、Hoechst 33258荧光染色法和流式细胞术检测不同浓度的二氢杨梅素溶液对骨肉瘤细胞形态、凋亡和周期的影响；并进一步采用Western Blot法分析细胞周期蛋白和凋亡蛋白表达的影响。研究结果表明二氢杨梅素对骨肉瘤细胞增殖具有显著的抑制作用，细胞形态发生了明显变化，呈显著的剂量依赖型，其半数抑制浓度（IC50）值为24.41±1.25 μM。细胞周期实验结果表明，经二氢杨梅素处理后，G0/G1期细胞数百分比从60.20%下降到21.50%，而G2/M细胞数百分比从11.60%增加到45.30%，细胞周期蛋白Cyclin B1表达下降引起细胞周期阻滞；此外，抗凋亡蛋白Bcl-2表达和促凋亡蛋白Bax表达的比例显著下降，从而诱导细胞发生凋亡。结果表明二氢杨梅素具有显著的抗骨肉瘤增殖活性，可开发一种潜在的治疗骨肉瘤的功能食品或药物。

In the present study, the inhibitory effect and mechanism of action of dihydromyricetin (DHM) on the proliferation of osteosarcoma cells were investigated. The inhibitory effects of DHM solutions at different concentrations on the proliferation of osteosarcoma cells were examined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. The influences of DHM solutions at different concentrations on the cell morphology, apoptosis and cell cycle of osteosarcoma cells were examined by an inverted microscope, Hoechst 33258 fluorescent staining and flow cytometry. The expression levels of cell cycle- and apoptosis-related proteins were analyzed by Western Blot method. The results showed that DHM exhibited a significant inhibitory effect on the proliferation of osteosarcoma cells and caused changes in the cell morphology of osteosarcoma cells in a dose-dependent manner, with the half inhibitory concentration (IC50) value as 24.41±1.25 μM. The results of cell cycle experiments showed that after the DHM treatment, the percentage of the cells at G0 phase to the cells at G1 phase (G0/G1) decreased significantly from 60.20% to 21.50%, whereas, the percentage of the cells at G2 phase to the cells at M phase increased from 11.60% to 45.30%, and the expression of cell cycle-related protein, Cyclin B1, decreased, causing the cell cycle arrest. In addition, the ratio of the expression of anti-apoptotic protein, Bcl-2, to the expression of pro-apoptotic protein, Bax, decreased, inducing apoptosis. These findings indicate that DHM has significant anti-proliferation activity against osteosarcoma cells, and may be a potential functional food or drug for the treatment of osteosarcoma.

广东大学生科技创新培育（“攀登计划”）专项资金项目（pdjh2019b0839)