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目的研究白藜芦醇(Res)对小鼠前脂肪细胞凋亡及细胞周期的影响及其机制。方法分别用25、50、75、100μmol/L的Res干预小鼠前脂肪细胞24、48、72 h,并设0μmol/L Res为阴性对照组(每组设3个复孔,重复3次)。全自动倒置荧光显微镜观察细胞的形态学变化、细胞增殖-毒性检测试剂盒检测细胞增殖活性、流式细胞仪检测细胞凋亡与细胞周期。结果形态学上,经50μmol/L Res干预48 h后的小鼠前脂肪细胞具有典型凋亡形态学改变。细胞增殖-毒性检测发现,0、25、50、75、100μmol/L Res干预24 h细胞增殖率分别为100%、(97.00±1.00)%、(91.00±2.65)%、(90.67±2.52)%和(86.00±3.61)%;干预48 h细胞增殖率分别是100%、(86.67±2.52)%、(76.00±2.00)%、(34.33±2.08)%和(30.33±2.52)%;干预72 h,细胞增殖率分别是100%、(82.00±2.65)%、(65.67±3.06)%、(21.00±3.61)%和(16.33±3.21)%。偏相关分析结果显示细胞增殖率与干预时间、Res浓度均呈负相关关系(r=-0.72、-0.83,均P<0.01)。0、25、50、75、100μmol/L Res干预24 h,G0/G1期的细胞比例分别为(27.23±2.63)%、(39.03±2.74)%、(80.20±5.15)%、(87.97±3.12)%和(90.80±2.08)%;S期的细胞比例分别为(72.43±2.99)%、(63.93±6.90)%、(19.80±5.15)%、(12.20±2.86)%和(9.20±2.08)%,2个细胞周期的50、75、100μmol/L Res干预组与阴性对照组之间的细胞比例差异均有统计学意义(P<0.05或P<0.01)。0、25、50、75、100μmol/L Res干预48 h,细胞凋亡率分别为(2.90±0.10)%、(5.40±3.81)%、(8.23±4.24)%、(29.77±6.18)%和(27.23±3.17)%;细胞坏死率分别为(7.50±0.87)%、(12.00±4.89)%、(12.27±3.81)%、(12.67±6.13)%和(20.73±2.64)%,100μmol/L的干预组的细胞调亡率、坏死率与阴性对照组的差异均具有统计学意义(P<0.05或P<0.01),其他各组与阴性对照组的差异均无统计学意义(P>0.05)。结论在一定的剂量范围内(0~100μmol/L),Res可能抑制小鼠前脂肪细胞生命周期并促进其凋亡。
Objective To investigate the effects of resveratrol (Res) on the apoptosis and cell cycle of mouse preadipocytes and its mechanism. Methods Preadipocytes were pretreated with 25, 50, 75 and 100 μmol / L of Res for 24, 48 and 72 h, respectively. Respectively, 0 μmol / L Res was used as a negative control group (3 replicate wells in each group, repeated 3 times) . The morphological changes of the cells were observed by inverted fluorescence microscope. The proliferation activity of the cells was detected by the kit of cytotoxicity and cytotoxicity. The apoptosis and the cell cycle were detected by flow cytometry. Results Morphologically, preadipocytes of mice with typical morphological changes of apoptosis were exposed to 50 μmol / L Res for 48 h. The results of cell proliferation and toxicity test showed that the proliferation rates of cells treated with 0, 25, 50, 75 and 100 μmol / L of Res for 24 h were (97.00 ± 1.00)%, (91.00 ± 2.65)% and (90.67 ± 2.52) And (86.00 ± 3.61)%, respectively. The cell proliferation rates at 48 h were 100%, (86.67 ± 2.52)%, (76.00 ± 2.00)%, (34.33 ± 2.08)% and The cell proliferation rates were 100%, (82.00 ± 2.65)%, (65.67 ± 3.06)%, (21.00 ± 3.61)% and (16.33 ± 3.21)%, respectively. Partial correlation analysis showed that the cell proliferation rate was negatively correlated with the time of intervention and the concentration of Res (r = -0.72, -0.83, all P <0.01). The percentage of cells in G0 / G1 phase was (27.23 ± 2.63)%, (39.03 ± 2.74)%, (80.20 ± 5.15)% and (87.97 ± 3.12) respectively at 0, 25, 50, 75 and 100μmol / ) And (90.80 ± 2.08)%, respectively. The percentage of cells in S phase were (72.43 ± 2.99)%, (63.93 ± 6.90)%, (19.80 ± 5.15)%, (12.20 ± 2.86)% and (9.20 ± 2.08) (P <0.05 or P <0.01). The percentage of cells in 50%, 75%, and 100μmol / L Res-treated and 2% of 2-cell cycles were significantly different. The apoptotic rate was (2.90 ± 0.10)%, (5.40 ± 3.81)%, (8.23 ± 4.24)% and (29.77 ± 6.18)% respectively at 0, 25, 50, 75 and 100μmol / (27.23 ± 3.17)% respectively. The cell necrosis rates were (7.50 ± 0.87)%, (12.00 ± 4.89)%, (12.27 ± 3.81)%, (12.67 ± 6.13)% and (20.73 ± 2.64)%, (P <0.05 or P <0.01). There was no significant difference between the other groups and the negative control group (P> 0.05) ). Conclusion Res may inhibit the lifespan and promote apoptosis of preadipocytes in a certain dose range (0 ~ 100μmol / L).