合江佛手挥发油调控Bcl-2/Bax蛋白表达诱导人恶性胶质瘤U87细胞凋亡  

Volatile oil of Hejiang Bergamot induces apoptosis through regulating protein expression of Bcl-2/Bax in human malignant glioma cell line U87 cells

作　　者：黄健麟 张丹 李颖 杨小会 王健 刘明华 HUANG Jian-lin;ZHANG Dan;LI Yin;YANG Xiao-hui;WANG Jian;LIU Ming-hua(School of Pharmacy,Southwest Medical University,Sichuan Luzhou 646000,China;Department of Pharmacy,Naxi District People's Hospital,Sichuan Luzhou 646000,China)

机构地区：[1]西南医科大学药学院,四川泸州646000 [2]泸州市纳溪区人民医院药剂科,四川泸州646000

出　　处：《中国医院药学杂志》2021年第7期681-686,共6页Chinese Journal of Hospital Pharmacy

摘　　要：目的:研究合江佛手挥发油诱导人恶性胶质瘤U87细胞凋亡及对凋亡相关因子Bcl-2和Bax蛋白表达的影响。方法:采用不同浓度的合江佛手挥发油作用于人乳腺癌细胞MCF-7、人恶性胶质瘤细胞U87、人肝癌细胞HepG2、人肺癌细胞A549,以CCK-8法(Cell Counting Kit-8 assay)筛选体外抗肿瘤活性最强的细胞;Hoechst33342/PI、透射电子显微镜观察及Annexin V-FITC/PI双染法相结合检测U87细胞的凋亡;Western Blot检测U87细胞凋亡相关因子Bcl-2、Bax蛋白表达的影响。结果:合江佛手挥发油对MCF-7、U87、HepG2、A549细胞的IC50分别为(173.99±18.68)、(82.19±13.07)、(192.87±19.66)和(215.83±23.27)μg·mL-1,其中对U87细胞抑制活性最强。合江佛手挥发油作用于U87细胞48 h后,Hoechst33342/PI、透射电子显微镜观察均可见核固缩、碎裂和凋亡小体等凋亡形态学改变。Annexin V-FITC/PI双染法结果显示,合江佛手挥发油和阳性对照维甲酸组U87细胞凋亡率明显增高,与阴性对照比较,P<0.05或P<0.01,药物作用呈现出较好的剂量依赖关系。Western Blot法结果表明,Bax蛋白表达增多,Bcl-2蛋白表达减少,Bcl-2/Bax比值降低。结论:合江佛手挥发油通过下调Bcl-2/Bax蛋白表达的比值,明显诱导人恶性胶质瘤细胞U87细胞凋亡。OBJECTIVE To explore the effect of volatile oil from Hejiang Bergamot(VO-HB) on apoptosis of human glioma cell line U87 and the expression of apoptosis related factors Bcl-2 and Bax.METHODS Different concentrations of VO-HB were utilized for MCF-7, U87, HepG2 and A549 cells. And the cells with the strongest anti-tumor activity were screened by CCK-8 assay. Hoechst33342/PI, transmission electron microscopy(TEM) and annexin V-FITC/PI double staining were employed for detecting apoptosis in U87 cells;Western Blot was used for detecting the expression of related factors Bcl-2 and Bax.RESULTS The IC50 values of VO-HB for MCF-7, U87, HepG2, A549 were(173.99±18.68),(82.19±13.07),(192.87±19.66) and(215.83±23.27) μg·mL-1 respectively. Thus the activity to U87 cells was the strongest in vitro. After 48-hour treatment of VO-HB, apoptotic morphological changes such as nuclear pyknosis, fragmentation and apoptotic bodies were observed by hoechst33342/PI and TEM. The results of annexin V-FITC/PI double staining showed that the apoptotic rate of U87 cells in VO-HB and positive control retinoic acid group significantly increased as compared with control(P<0.05 or P<0.01), showing an excellent dose-dependent relationship. Western blot showed that Bax protein expression increased, bcl-2 protein expression decreased and Bcl-2/Bax ratio declined.CONCLUSION VO-HB may significantly induce the apoptosis of human malignant glioma cell line U87 by down-regulating the protein expression ratio of Bcl-2/Bax.

关 键 词：合江佛手挥发油 U87 凋亡 BCL-2 BAX 

分 类 号：R969[医药卫生—药理学]