槲皮素对大鼠脑胶质瘤C6细胞增殖调控的作用

Effect of quercetin on proliferation of rat glioma C6 cells

目的:研究类黄酮化合物槲皮素(quercetin,QUE)对体外培养的大鼠脑胶质瘤C6细胞增殖调控的作用。方法:按QUE浓度分成10、25、50、75及100μmol.L-15个处理组和空白对照组(生理盐水)及溶剂对照组(二甲亚砜),大鼠脑胶质瘤C6细胞在RPMI 1640培养基中生长达1×106.mL-1后,在96孔板中分别加入上述浓度的QUE继续培养,每组设3复孔,作用24、48及72 h,采用MTT比色法检测QUE对大鼠脑胶质瘤C6细胞的增殖抑制情况,流式细胞术(FCM)对50及100μmol.L-1的QUE作用48 h的大鼠脑胶质瘤C6细胞进行周期分析,免疫组化法检测50μmol.L-1的QUE作用48 h的p53和bcl-2基因产物。结果:与空白对照组比较,QUE处理组随药物浓度增加和作用时间的延长,A值减小(P<0.05),对C6细胞的增殖抑制作用增强,使停滞在G0/G1期的细胞增加(P<0.01),而S和G2/M期细胞减少(P<0.05)。P53蛋白表达增加和Bcl-2蛋白表达减少。结论:QUE对C6细胞增殖抑制作用具有浓度、时间依赖性,通过P53蛋白表达增加和Bcl-2蛋白表达减少诱导细胞凋亡来实现。

Objective To study the effects of quercetin （QUE） on proliferation of rat glioma C6 cell line in vitro. Methods The cells were divided into 5 treatment groups （10, 25, 50, 75 and 100 μmol·L^-1 QUE）, blank control and menstruum control group. The rat C6 cells were cultivated to 1× 10^6· mL^-1 in the RPMI 1640 medium, then added into 96 holes board with various doses of QUE by 3 holes per group, and MTT assay was used to observe the proliferation of the cells treated for 24, 48 and 72 h. The change of cell cycle was also observed by flow cytometry （FCM） after the cells were treated with 50 and 100μmol·L^-1 QUE for 48 h. The changes of the protein P53 and Bcl-2 of C6 cells treated with 50 /μmol·L^-1 QUE for 48 h were detected by immunocytochemical methods. Results With the augmentation of QUE and the extension of the treated time, the C6 cell growth was inhibited, the A values decreased and the cell number in G0/C-~ phase was increased, the cell numbers in S and G2/ M phases were cut down, and the decreased expression of Bcl-2 protein and the increased expression of P53 protein were also observed after treatment with QUE. Conclusion Inhibitory effect of QUE on C6 cell line is proved to be dependent on the treated time of the drug and the dose of QUE, and the induced apoptosis of C6 cells is implemented by the means of up-regulation of P53 protein expression and down-regulation of Bcl-2 protein expression.