摘要
目的探讨高压氧(HBO)联合替莫唑胺(TMZ)对神经胶质瘤U373MG细胞凋亡的影响及其作用机制.方法U373MG细胞培养完成后分为4组:对照组(不作HBO及药物处理)、TMZ处理组(只给予50μmol/L TMZ处理)、HBO组(0.24 MPa HBO处理3 h)和HBO+TMZ处理组(HBO预处理3 h后加入50μmol/L TMZ).CCK-8法检测HBO和TMZ单用或联用对U373MG细胞增殖的影响;流式细胞术检测HBO和TMZ单用或联用对U373MG细胞情况及细胞中活性氧(ROS)水平变化情况;Western blotting实验检测U373MG细胞凋亡相关蛋白的表达.结果HBO+TMZ组中U373MG细胞的增殖抑制情况较TMZ组更明显(P<0.05);TMZ处理组中U373MG细胞出现了早期凋亡和晚期凋亡,HBO+TMZ组中凋亡率明显高于对照组[(73.19±3.58)%vs.(30.5±2.27)%,P<0.01].与空白组和HBO组比较,TMZ组中Bcl-2表达量明显降低,Bax、caspase-3表达量明显升高,且HBO+TMZ组较TMZ组变化更明显(P<0.05或P<0.01).HBO+TMZ组U373MG细胞中ROS水平明显升高(P<0.01);加入ROS清除剂N-乙酰半胱氨酸(NAC)后,HBO+TMZ组的细胞凋亡被抑制(P<0.01).结论HBO联合TMZ能够增强对U373MG细胞增殖的抑制作用,并通过上调节U373MG细胞中的ROS水平促进TMZ诱导细胞凋亡.
Objective To investigate the effect of hyperbaric oxygen(HBO)combined with temozolomide(TMZ)on the apoptosis of glioma U373MG cells and also to explore possible mechanism involved.Methods Upon completion of cell culture,they were divided into 4 groups,i.e.the control group,the TMZ group,the HBO group and the HBO+TMZ group.The control group received neither HBO nor drug treatment,the TMZ group only received 50μmol/L TMZ,the HBO group just received 0.24 MPa HBO treatment for 3 hours and the HBO+TMZ group was given 50μmol/L TMZ 3 hours after HBO pretreatment.CCK-8 method was used to detect the effects of HBO and TMZ alone or combined use of 2 treatment methods on the proliferation of glioma U373MG cells.Flow cytometry was used to detect HBO and TMZ alone or combined use of 2 treatment methods on glioma U373MG cells and changes in the level of reactive oxygen species(ROS).Western blotting was used to detect the expression of U373MG apoptosis-related proteins.Results The proliferation inhibition of U373MG cells in the HBO+TMZ group was more obvious as compared with that of the TMZ group(P<0.05).U373MG cells in the TMZ group showed early and late apoptosis,the rate of apoptosis in the HBO+TMZ group was significantly higher than that of the control group[(73.19±3.58)%vs.(30.5±2.27)%](P<0.01).As compared with the blank control and HBO groups,the expression level of Bcl-2 in the TMZ group significantly decreased,while the expression level of Bax and caspase-3 markedly increased.Furthermore,more obvious changes could be detected,when the HBO+TMZ group was compared with the TMZ group(P<0.05 or P<0.01).ROS level in the U373MG cells of the HBO+TMZ group obviously increased(P<0.01).With the addition of ROS scavenger NAC,apoptosis was inhibited in the HBO+TMZ group(P<0.01).Conclusion HBO combined with TMZ could enhance the inhibitory effect of TMZ on the proliferation of U373MG cells,and promote apoptosis induced by TMZ through up-regulating ROS levels in U373MG cells.
作者
马建功
王晓斌
侯欣
李明轩
方树民
刘亚虎
何承
Ma Jiangong;Wang Xiaobin;Hou Xin;Li Mingxuan;Fang Shumin;Liu Yahu;He Cheng(Department of Neurosurgery,First Affiliated Hospital of Henan University,Kaifeng 475000,China;不详)
出处
《中华航海医学与高气压医学杂志》
CAS
CSCD
2019年第5期404-407,共4页
Chinese Journal of Nautical Medicine and Hyperbaric Medicine
基金
河南省教育厅科学技术研究重点项目(16A320072)。
