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目的:探讨虎纹捕鸟蛛毒抑制体外脑胶质瘤细胞系U251生长并诱导其凋亡,激活半胱天冬酶-3(Caspase-3)的作用。方法:将U251细胞分为3组:空白组、顺铂组、加药组(虎纹捕鸟蛛毒素浓度为37.5,50,75,100,150 mg·L-1),毒素作用24,48 h后用四甲基偶氮唑蓝(MTT)法检测细胞活性并与顺铂组、空白组做对比;流式细胞仪AnnexinV-FITC检测细胞凋亡率;Caspase-3分光光度法检测Caspase-3的相对活性。结果:虎纹捕鸟蛛毒素剂量为37.5,50,75,100,150 mg·L-1时,对U251细胞的增殖有显著性抑制作用(P<0.05或P<0.01),IC50为53.48 mg·L-1。虎纹捕鸟蛛毒素可诱导U251胶质瘤细胞凋亡并呈浓度依赖关系。在剂量为150 mg·L-1诱导48 h,细胞早期凋亡率84.1%,晚期凋亡率4.48%。用不同浓度分别处理细胞,Caspase-3活性于48 h,150 mg·L-1时达高峰,对细胞中的执行分子Caspase-3的激活速度快,活化程度为9.23,并存在剂量依赖关系和浓度依赖性的升高。结论:虎纹捕鸟蛛毒素明显抑制U251细胞生长并诱导其发生凋亡,凋亡过程中有Caspase-3的激活。
OBJECTIVE: To investigate the effect of Huwenthuis poisoning on inhibiting the growth of human glioma cell line U251 and inducing its apoptosis and activating Caspase-3. Methods: U251 cells were divided into three groups: blank group, cisplatin group, dosing group (tigerwort toxin concentration of 37.5,50,75,100,150 mg · L-1), toxins 24,48 h after four Methyl thiazolyl tetrazolium (MTT) assay was used to detect cell viability and compared with cisplatin group and blank group; apoptosis rate was detected by flow cytometry Annexin V-FITC; Caspase-3 spectrophotometry was used to detect the relative activity of Caspase-3 . Results: The dosages of tiger beetle were 37.5, 50, 75, 100 and 150 mg · L-1, respectively, and significantly inhibited the proliferation of U251 cells (P <0.05 or P <0.01) with IC50 of 53.48 mg · L -1 . Huwentuctoxin can induce apoptosis in U251 glioma cells in a concentration-dependent manner. At the dose of 150 mg · L-1 for 48 h, the early apoptosis rate was 84.1% and the late apoptosis rate was 4.48%. Caspase-3 activity peaked at 48 h and 150 mg · L-1, and the activation rate of Caspase-3 was 9.23 in a dose-dependent manner Concentration-dependent increase. Conclusion: Huwentoxin obviously inhibits the growth of U251 cells and induces its apoptosis. Caspase-3 activation is observed during apoptosis.