摘要
目的:研究卫矛醇对C6胶质瘤细胞株的体外增殖及凋亡的影响。方法:将C6大鼠胶质瘤细胞随机分为对照组、卫矛醇干预组、卫矛醇+雷帕霉素组、卫矛醇+3-MA组,各组予不同药物孵育,采用MTT法检测卫矛醇对C6细胞的体外增殖抑制率,筛选出抑制率相对较高且生长状态良好的细胞;应用Hoechest 33342染色透射电镜观察细胞凋亡亚显微结构的改变,采用免疫荧光和Western Blot技术检测与凋亡和自噬相关蛋白的表达水平。结果:卫矛醇对C6胶质瘤细胞增殖有明显的抑制作用,并呈现出良好的浓度-效应相关性,与对照组比较,随着浓度的增加,细胞的增殖抑制率上升(P<0.01);Hoechst染色结果显示,与对照组相比,共同孵育卫矛醇24 h后的C6细胞出现明显的核固缩现象,证明有大量的细胞出现凋亡;Western Blot结果显示,与对照组相比,促凋亡基因Bax表达上调,抗凋亡基因Bcl-2表达下调。结论:卫矛醇能够抑制C6细胞的增殖,并诱导其凋亡,其作用机制可能与上调Bax、下调Bcl-2有关。
Objective:To investigate the effect of dulcitol on the in vitro proliferation and apoptosis of C6 glioma cell line.Methods:Rat C6 glioma cells were randomly divided into control group,dulcitol intervention group,dulcitol+rapamycin group,and dulcitol+3-MA group,and these groups were incubated with different drugs.MTT assay was used to measure the inhibition rate of dulcitol on the in vitro proliferation of C6 cells,and the cells with a relatively high inhibition rate and good growth status were obtained;Hoechest33342 staining and transmission electron microscopy were used to observe the change in cell apoptosis and submicroscopic structure;immunofluorescence assay and Western blot were used to measure the expression levels of apoptosis-and autophagy-related proteins.Results:Dulcitol showed a marked inhibitory effect on the proliferation of C6 glioma cells,with a good concentration-effect relationship,and compared with the control group,the inhibition rate of cell proliferation increased with the increase in concentration(P<0.01).Hoechst staining showed that compared with the control group,C6 cells incubated with dulcitol for 24 hours had marked karyopyknosis,suggesting the apoptosis of a large number of cells.Western blot showed that compared with the control group,there was an increase in the expression of the pro-apoptotic gene Bax and a reduction in the expression of the anti-apoptotic gene Bcl-2.Conclusion:Dulcitol can inhibit the proliferation of C6 cells and induce the apoptosis of C6 cells,possibly by upregulating Bax and downregulating Bcl-2.
作者
张玉岭
张涛
陈宝贵
王晖
ZHANG Yuling;ZHANG Tao;CHEN Baogui;WANG Hui(Beichen Hospital of Traditional Chinese Medicine,Tianjin 300499,China;School of Life Sciences,Nankai University,Tianjin 300071,China;Wuqing Hospital of Traditional Chinese Medicine,Tianjin 301799,China)
出处
《湖南中医杂志》
2024年第2期168-172,200,共6页
Hunan Journal of Traditional Chinese Medicine
基金
天津市卫生健康委中医、中西医结合科研项目(2023123)
第二批卫生健康行业高层次人才选拔培养工程(TJSJMYXYC-D2-057)。