摘要
目的:探讨小檗胺体外对大鼠骨肉瘤UMR-106细胞增殖和凋亡的作用及机制。方法:用不同浓度的小檗胺(0,2,4,8,16,32mg.L-1)分别处理UMR-106细胞,作用时间分别为24,48,72h,采用四甲基偶氮唑盐(MTT)比色法检测小檗胺对UMR-106细胞增殖的抑制作用。用不同浓度的小檗胺(0,4,8,16mg.L-1)分别处理UMR-106细胞24h后,采用流式细胞术(FCM)检测细胞凋亡率(Annexin/PI染色),采用酶联免疫吸附试验(ELISA)检测细胞内Bcl-2、Bax水平,采用分光光度法检测caspase-3的活性。结果:小檗胺对UMR-106细胞的作用呈时间-剂量依赖性,孵育72h、32mg.L-1的小檗胺对UMR-106细胞的作用最强,抑制率接近90%;24,48,72h的IC50分别为24.69、8.03和3.54mg.L-1。空白对照组和小檗胺处理组细胞凋亡率分别为(1.64±0.29)%,(3.58±0.31)%,(6.3±0.5)%和(11.3±1.1)%。同时,小檗胺可以诱导细胞坏死;此外,小檗胺剂量依赖性地降低Bcl-2,增高Bax,降低Bcl-2/Bax比值,并增强caspase-3的活性。结论:小檗胺体外能抑制UMR-106细胞增殖并诱导细胞凋亡或坏死,其诱导凋亡的机制可能与降低Bcl-2/Bax比值,增强caspase-3的活性有关。
OBJECTIVE To explore the effect of berbamine on the proliferation and apoptosis of rat osteosarcoma UMR-106 cells and its possible mechanism in vitro.METHODS Rat osteosarcoma UMR-106 cells were treated by berbamine with different concentrations（0,2,4,8,16,32 mg·L-1） for 24 h,48 h and 72 h,respectively.The anti-proliferation was assayed by methyl thiazolyl tetrazolium（MTT）.Rat osteosarcoma UMR-106 cells were cultured with different concentrations of berbamine（0,4,8,16 mg·L-1） for 24 h,the apoptotic rate was detected by flow cytometry using Annexin V/PI labeled.The levels of Bcl-2 and Bax were measured by ELISA and the activity of caspase-3 was detected by spectrophotometry.RESULTS The inhibitory rate of cell proliferation increased in a time-and dose-dependent manner,and the highest inhibitory rate almost arrived at 90% when UMR-106 cells were treated by berbamine 32 mg·L-1 for 72 h.The IC50 for 24,48 and 72 h were 24.69,8.03 and 3.54 mg·L-1,respectively.Apoptotic rates of control group and berbamine treated group were（1.64±0.29）%,（3.58±0.31） %,（6.3±0.5） % and（11.3±1.1） %,respectively.Meanwhile,berbamine could induce necrosis.In addition,berbamine dose-dependently reduced Bcl-2 level and increased Bax,causing the reduction of the Bcl-2/Bax ratio and increasing the caspase-3 activity in UMR-106 cells significantly.CONCLUSION Berbamine can inhibit the proliferation and induce apoptosis or necrosis of UMR-106 cells in vitro,the reducing Bcl-2/Bax ratio and increasing the caspase-3 activity might contribute to the apoptotic mechanisms.
出处
《中国医院药学杂志》
CAS
CSCD
北大核心
2012年第21期1719-1722,共4页
Chinese Journal of Hospital Pharmacy
