汉黄芩素及其抑制物细胞周期蛋白依赖性激酶1对人脑胶质瘤干细胞作用机制的研究

A research of mechanism and effects of wogonin and its inhibitor cyclin-dependent kinase 1 on human glioma stem cells

目的研究汉黄芩素及其抑制剂细胞周期蛋白依赖性激酶1（CDKl）对人脑胶质瘤干细胞（BGSCs）的作用机制。方法使用不同浓度的汉黄芩素（1、10、1000μmol／L）对BGSCs进行浸染，并设空白对照组。采用四甲基偶氮唑盐（M1fr）比色法检测汉黄芩素对BGSCs增殖的影响；采用逆转录一聚合酶链反应（RT—PCR）检测CDKlmRNA表达；同时检测脂质过氧化产物还原型谷胱甘肽（GSH）、超氧化物歧化酶（SOD）和丙二醛（MDA）含量的变化。结果汉黄芩素对人BGSCs增殖具有明显的抑制作用，各剂量汉黄芩素组BGSCs增殖[吸光度（A）值]较空白对照组明显降低，随给药浓度增加，细胞生长抑制率（IR）较空白对照组逐渐增加，以汉黄芩素100tanol／L组作用更显著[A值：0．18±0．02比0．67±0．03，IR：（64．43±0．04）％比0，均P〈0．05]；各剂量汉黄芩素组CDKlmRNA表达（A值）均较空白对照组有不同程度的提高，在10tanol／L、100~mol／L汉黄芩素组出现显著提高（0．378±0．061、0．733±0．081比0．125±0．019，均P〈0．05）；脂质过氧化产物GSH（mg／g）、SOD（U／g）和MDA（nmol／g）在各个剂量汉黄芩素组与空白对照组比较差异均无统计学意义（GSH：474．51±38.63、479．67±15．89、481．17±36．41比487．00±28．53．SOD：26．58±2．26、26．75±1．96、27．08±1．52比27．31±1．89，MDA：4．92±0．45、4．72±0．73、4．61±0．47比4．45±0．26，均P〉0．05）。结论CDKl可能参与汉黄芩素抑制人BGSCs生长的作用机制，该作用与脂质过氧化产物无关。

Objective To investigate the meclaanism and effects of wogonin and its inhibitor cyclin- dependent kinase 1 （CDK1） on human brain glioma stem cells （BGSCs）. Methods BGSCs were treated with different concentrations （ 1,10,100 μmol/L） of wogonin and a blank control group was made. Cell growth inhibition of BGSCs was detected by assay of methyl thiazolyl tetrazolium （MTT）. Expression of CDK1 mRNA was detected by retro transcriptive-polymerase chain reaction （ RT-PCR）. Simultaneously, lipid peroxidation products of glutathione （ GSH ）, superoxide dismutase （SOD）, malondiadehyde （MDA） were tested. Results Wogonin significantly suppressed the growth of BGSCs [ absorbance （A） value ], statistical significant differences were found in study groups compared with blank control group. The cell growth inhibition rate （IR） was gradually increased with the administration of wogonin concentration increment, especially prominent in the wogonin 100 μmol/L group （A value ： 0.18±0.02 vs. 0.67 ＋ 0.03, IR ： （64.43 ＋ 0.04） % vs. 0, both P〈 0.05）. Expression of CDK1 mRNA was significantly increased in 10 tmaol/L and 100 μmol/L wogonin groups compared to that in the blank control group （0.378 ＋0.061,0.733-4-0.081 vs. 0.125 ＋0.019, both P〈0.05）. There was no significant change in GSH （mg/g）, SOD （U/g） and MDA （nmol/g） in all groups （GSH： 474.51＋38.63, 479.67＋15.89, 481.17＋36.41 vs. 487.00＋28.53, SOD： 26.58＋2.26, 26.75 ＋ 1.96,27.08 -4- 1.52 vs. 27.31 ＋ 1.89, MDA ： 4.92 ＋ 0.45,4.72 ＋ 0.73,4.61 ＋ 0.47 vs. 4.45 ＋ 0.26, all P 〉 0.05 ）. Conclusion CDK1 possibly participates in the mechanism of wogonin suppression of BGSCs growth which is not related to lipid oeroxidation.