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目的:研究茶多酚对人多发性骨髓瘤RPMI 8226细胞生长及凋亡的影响,并探讨其可能的作用机制。方法:通过MTT法、瑞氏染色法和FCM法分别检测不同浓度的茶多酚对RPMI 8226细胞增殖和凋亡的影响;FCM法检测对细胞线粒体膜电位的影响;蛋白质印迹法检测Δcaspase-3片段和凋亡相关蛋白bcl-2的表达;ELISA法检测对RPMI 8226细胞分泌细胞因子白细胞介素-6(interleukin-6,IL-6)水平的影响。结果:与对照组相比,茶多酚浓度129.6和259.1μmol/L处理RPMI 8226细胞1～3d后均能明显抑制细胞的增殖,且呈时间-剂量相关性;统计显示,24h时的半数细胞抑制浓度(half inhibitory concentration,IC50)值为165.7 μmol/L。茶多酚(129.6 μmol/L)作用24h后,RPMI 8226细胞出现典型的凋亡形态学改变;茶多酚(129.6和259.1 μmol/L)作用24h后,RPMI 8226细胞凋亡率分别为(24.6±2.3)%和(43.5±3.9)%,与对照组比较差异均有统计学意义;细胞的线粒体膜电位降低,活性Δcaspase-3片段产生,bcl-2蛋白表达水平下降,IL-6分泌水平降低。结论:茶多酚能抑制RPMI 8226细胞增殖并诱导其凋亡,其作用机制可能与产生活性Δcaspase-3片段,抑制bcl-2蛋白表达,降低细胞分泌IL-6水平相关。
Objective: To study the effect of tea polyphenols on the growth and apoptosis of human multiple myeloma RPMI 8226 cells and to explore its possible mechanism. Methods: The effects of different concentrations of tea polyphenols on the proliferation and apoptosis of RPMI 8226 cells were detected by MTT assay, Wright’s stain and FCM. The effects of FCM on the mitochondrial membrane potential were detected by Western blotting. The expressions of Δcaspase-3 The expression of bcl-2 was detected by enzyme-linked immunosorbent assay (ELISA). The effect of interleukin-6 (IL-6) on RPMI 8226 cells was detected by ELISA. Results: Compared with the control group, the proliferation of RPMI 8226 cells was inhibited at the concentration of 129.6 and 259.1μmol / L for 1 ~ 3d, and the time-dose dependence was observed. The statistics showed that half cells The half inhibitory concentration (IC50) value was 165.7 μmol / L. Apoptosis of RPMI 8226 cells was observed after treated with tea polyphenols (129.6 μmol / L) for 24 h. Apoptotic rates of RPMI 8226 cells treated with tea polyphenols (129.6 and 259.1 μmol / L) for 24 h were (24.6 ± 2.3% and (43.5 ± 3.9)%, respectively, which were significantly different from the control group. The mitochondrial membrane potential of the cells was decreased, the activity of Δcaspase-3 fragment was decreased, the expression of bcl-2 protein and the secretion of IL-6 reduce. CONCLUSION: Tea polyphenols can inhibit the proliferation and induce apoptosis of RPMI 8226 cells. The mechanism may be related to the production of active Caspase-3 fragment, the inhibition of bcl-2 protein expression and the decrease of IL-6 secretion.