摘要
目的探讨五味子多糖对人脑胶质瘤U251细胞体外生长的抑制作用及诱导细胞凋亡的作用机制。方法实验分为4组,对照组常规培养(不加任何药物)人脑胶质瘤U251细胞,五味子多糖250μg/mL、500μg/mL、800μg/mL组分别加入相应浓度的五味子多糖培养人脑胶质瘤U251细胞。采用MTT法检测各组细胞培养12,24,48 h后生长抑制率;Transwell细胞体外侵袭实验法使细胞穿过小室透膜数量,检测各组细胞迁移能力;流式细胞实验结合AnnexinV-FITC/PI双染色法检测各组细胞培养12,24,48 h后早期凋亡百分比。RT-PCR技术检测500μg/mL五味子多糖作用U251细胞12,24,48 h后bcl-2、bax、caspase-3基因表达水平。结果与对照组比较,不同浓度五味子多糖对U251细胞有显著增殖抑制作用(P均<0.05),且抑制作用呈现剂量与时间效应关系;与对照组比较,五味子多糖各组细胞迁移抑制率、细胞早期凋亡率、细胞晚期凋亡率均明显增高(P均<0.05),且细胞迁移抑制率和细胞凋亡率随五味子多糖浓度增高而增高(P均<0.05)。500μg/mL五味子多糖作用U251细胞12,24,48 h后,Bax、Caspase-3基因表达水平及Bax/Bcl-2比值逐渐增高,Bcl-2基因表达水平逐渐降低,与对照组比较差异均有统计学意义(P均<0.05)。结论五味子多糖能显著抑制人脑胶质瘤U251细胞生长,诱导细胞凋亡,降低细胞迁移能力,其促进凋亡作用机制可能与上调Bax、Caspase-3基因表达水平,下调Bcl-2基因表达水平相关。
Objective It is to investigate the inhibitory effect of schisandra chinensis polysaccharides on the growth of human glioma U251 cells in vitro and the mechanism of inducing apoptosis.Methods The experiment was divided into 4 groups,the human glioma U251 cells were routinely cultured(without any drugs)in the control group,and were cultured with the corresponding concentration of schisandra polysaccharide in 250μg/mL,500μg/mL,800μg/mL schisandra polysaccharide groups.MTT method was used to detect the growth inhibition rate of cells after 12,24,and 48 hours of culture in each group;Transwell cell invasion test method was used to detect the number of cells through the small room of membrane to evaluate the cell migration ability in each group;flow cytometry combined with AnnexinV-FITC/PI Double staining method was used to detect the percentage of early apoptosis after 12,24,and 48 hours of cell culture in each group.RT-PCR technique was used to detect the expression levels of Bcl-2,Bax and Caspase-3 genes after U251 cells were treated with 500μg/mL schisandra polysaccharides for 12,24 and 48 h.Results Compared with the control group,different concentrations of schisandra chinensis polysaccharides had significant proliferation inhibitory effect on U251 cells(all P<0.05),and the inhibitory effect showed a dose-time effect relationship;compared with the control group,the cell migration inhibition rate,early cell apoptosis rate and late cell apoptosis rate were significantly increased in every schisandra chinensis polysaccharides group(P<0.05),and the cell migration inhibition rate and apoptosis rate increased with the increase of Schisandra chinensis polysaccharide concentration(P<0.05).After the treatment with 500μg/mL schisandra chinensis polysaccharides for 12,24,and 48 hours,the expression levels of Bax and Caspase-3 gene and Bax/Bcl-2 ratios gradually increased,and Bcl-2 gene expression levels gradually decreased,the differences were statistically significant compared with control group(all P<0.05).Conclusion Schisandra chinensis polysaccharides can significantly inhibit the growth of human glioma U251 cells,induce apoptosis,and reduce cell migration ability.The mechanism of promoting apoptosis may be related to up-regulation of Bax and Caspase-3 gene expression levels and down-regulation of Bcl-2 gene expression levels.
作者
王光杰
张熙
WANG Guangjie;ZHANG Xi(The People’s Hospital Affiliated to Hubei Medical College, Shiyan 442000, Hubei, China)
出处
《现代中西医结合杂志》
CAS
2020年第18期1968-1972,共5页
Modern Journal of Integrated Traditional Chinese and Western Medicine
作者简介
王光杰,男,硕士,研究方向为神经痛机制及治疗;通讯作者:张熙,E-mail838624729@qq.com。
