摘要
目的观察仙茅总黄酮诱导人骨肉瘤MG-63细胞凋亡并探讨其可能的机制。方法采用加热回流提取和大孔吸附树脂纯化方法制备仙茅总黄酮。通过CCK-8法检测MG-63细胞的增殖抑制率,计算半数抑制浓度(IC50);Hoechst 33258染色法观察凋亡细胞核形态;流式细胞术检测细胞凋亡率;Western blotting法和RT-PCR法检测JNK、p38 MAPK、Bax、Bcl-2和Caspase-3的蛋白和mRNA表达水平。结果纯化后仙茅总黄酮纯度为40.72%;与空白对照组相比,仙茅总黄酮对MG-63细胞的增殖抑制作用明显,且呈时间和剂量依赖性;药物组凋亡率显著升高;Western blotting及RT-PCR结果显示,随药物浓度升高,JNK、p38 MAPK和Bcl-2的蛋白和mRNA表达水平均显著降低,Caspase-3、Bax的蛋白和mRNA表达水平均显著升高(P<0.05,P<0.01或P<0.001)。结论仙茅总黄酮能显著抑制人骨肉瘤MG-63细胞增殖、促进其凋亡,其作用机制可能与下调MAPK信号通路JNK、p38 MAPK及抗凋亡因子Bcl-2的表达,上调Caspase-3和促凋亡因子Bax的表达有关。
Objective:To investigate the effect of Curculio Orchioides total flavonoids on human osteosarcoma MG-63 cells apoptosis.Methods:The total flavonoids were prepared by heating reflux extraction and macroporous adsorption resin purification methods.CCK-8 method was used to detect its anti-proliferative activity on MG-63 cells,and the half inhibitory concentration(IC50)was calculated.Hoechst 33258 staining method was used to observe the nucleus morphology of apoptotic cells.The protein and mRNA levels of JNK,p38 MAPK,Bax,Bcl-2and Caspase-3 were detected by Western blotting and RT-PCR.Results:The purity of the total flavonoids of Curculio Orchioides after purification was 40.72%;compared with the control group,the results showed that Curculio Orchioides total flavonoids had a significant inhibitory effect on the proliferation of MG-63 cells in a time and dose-dependent manner.The apoptosis rate was increased significantly in the drug group.As the drug concentration increased,Western blotting and RT-PCR results showed that the protein and mRNA levels of JNK,p38MAPK and Bcl-2 were reduced,and the protein and mRNA expression levels of Caspase-3 and Bax were increased significantly(P<0.05 or P<0.01 or P<0.001).Conclusion:Curculio Orchioides total flavonoids can significantly inhibit the proliferation and migration of human osteosarcoma MG-63 cells and promote their apoptosis.The mechanism of action may be related to down-regulating the expression of MAPK signaling pathway JNK,p38MAPK and anti-apoptotic factor Bcl-2,and up-regulating the expression of Caspase-3 and pro-apoptotic factor Bax.
作者
李婷
汪小玉
税丕先
石厚银
张旗
沈骅睿
Li Ting;Wang Xiaoyu;Shui Pixian;Shi Houyin;Zhang Qi;Shen Huarui(Southwest Medical University,Sichuan,Luzhou 646000,China)
出处
《中国中医急症》
2023年第9期1529-1533,共5页
Journal of Emergency in Traditional Chinese Medicine
基金
四川省科技计划联合创新专项(泸州)(2022YFS0609)
泸州市人民政府—西南医科大学科技战略合作项目(2021LZXNYD-J31)
四川省中医药管理局中医药科研专项课题(2021MS463)
西南医科大学—西南医科大学附属中医医院联合专项(2020XYLH-013)。
关键词
骨肉瘤
MG-63细胞
仙茅总黄酮
凋亡
机制
Osteosarcoma
MG-63 cells
Curculio Orchioides total flavonoids
Apoptosis
Mechanism
