白藜芦醇脂质体对C6胶质瘤细胞的抑制作用分析

Research for Inhibitory Effect of Resveratrol Liposomes on C6 Glioma Cells

目的:制备白藜芦醇脂质体并对其体外抗肿瘤作用进行评价。方法:通过薄膜分散-硫酸铵梯度法制备白藜芦醇脂质体,使用粒度仪对脂质体进行表征;确定C6胶质瘤细胞对数生长期;通过磺酰罗丹明B蛋白(SRB)法评价游离白藜芦醇及其脂质体对C6胶质瘤细胞的抗增殖作用,利用流式法考察白藜芦醇及其脂质体对C6胶质瘤的凋亡作用,以及C6胶质瘤细胞对白藜芦醇和其脂质体的摄取作用。结果:白藜芦醇脂质体的平均粒径(139.97±0.64)nm,Zeta电位(-7.00±0.74)m V;游离白藜芦醇和白藜芦醇脂质体对C6胶质瘤细胞的抑制率最高分别可达(73.30±0.56)%和(91.70±0.60)%,差异有统计学意义(P<0.05);游离白藜芦醇和白藜芦醇脂质体对C6胶质瘤细胞的诱导凋亡率分别为(20.03±0.85)%和(27.18±0.96)%,差异有统计学意义(P<0.05);C6胶质瘤细胞对游离白藜芦醇和白藜芦醇脂质体的摄取率分别为(67.79±1.19)%和(77.61±1.67)%。结论:相比于游离的白藜芦醇,白藜芦醇脂质体对C6胶质瘤细胞的抗增殖作用更明显,具有更强的诱导C6胶质瘤细胞凋亡作用。白藜芦醇脂质体可以更多地被C6胶质瘤细胞摄取,这在一定程度上促进了其对C6胶质瘤细胞的抗增殖作用和诱导凋亡能力。说明白藜芦醇脂质体具有较强的体外抗C6胶质瘤作用。

Objective： To prepare resveratrol liposomes and evaluate its anti-tumor effect in vitro.Method： The resveratrol liposomes were prepared by film dispersion-ammonium sulfate gradient method and the free resveratrol was filtered by sephadex G-50 column afterwards. Moreover,the particle size and Zeta potential of liposomes were characterized by laser particle size analyzer. C6 glioma cells were cultured for 4 days and their logarithmic growth phase was decided by cell growth curve. These cells were tested in several experiments in vitro as below. The antiproliferative effect of free resveratrol on C6 glioma cells were evaluated by sulforhodamine B（ SRB） method,while the antiproliferative effect of resveratrol liposomes on C6 glioma cells were tested by the same way. Furthermore,the apoptosis effect on C6 glioma cells treated by free resveratrol were tested by flow cytometry method. Also,the apoptosis effect on C6 glioma cells treated by resveratrol liposomes were observed by the same way. The uptake effect of resveratrol and resveratrol liposomes on C6 glioma cells were all determined by this method. Result： The particle size of blank liposomes and resveratrol liposomes were separately（ 144. 57 ±0. 93） nm and（ 139. 97 ± 0. 64） nm,while their Zeta potentials were respectively（-6. 70 ± 0. 93） m V and（-7. 00 ± 0. 74） m V. The logarithmic growth phase of C6 glioma cells was between 36 h to 48 h. The SRB method showed that the inhibitory effect on C6 glioma cells treated by resveratrol liposomes was（ 91. 70 ±0. 60） %,which was significantly higher than（ 73. 30 ± 0. 56） % of free resveratrol（ P〈0. 05）. The apoptosis effect on C6 glioma cells induced by resveratrol liposomes was（ 27. 18 ± 0. 96） %,which was obviously higher than（ 20. 03 ± 0. 85） % of free resveratrol（ P〈0. 05）. The uptake effect of resveratrol liposomes was（ 77. 61 ±1. 67） %,which was also significantly higher than（ 67. 79 ± 1. 19） % of free resveratrol（ P〈0. 05）.Conclusion： Compared with free resveratrol,resveratrol liposomes has a more significant antiproliferative effect against C6 glioma cells, and has a stronger apoptosis effect on C6 glioma cells. The number of resveratrol liposomes that uptake is obviously more than that of free resveratrol. It is concluded that the resveratrol liposomes built in this study shows strongest anti-tumor effect on C6 glioma cells in vitro by comparing with other controlled groups.