青葙苷A诱导肝癌HepG2细胞凋亡及相关机制研究  

Study on Apoptosis of HepG2 Cell Induced by Celosin A from Celosia Semen and its Mechanism

作　　者：程齐来[1,2] 李洪亮[1] 黄志勤[1] 

机构地区：[1]赣南医学院药学院,江西赣州341000 [2]湖南中医药大学,长沙410208

出　　处：《中国实验方剂学杂志》2013年第23期200-204,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：江西省自然科学基金项目(2012ZBAB205006);江西省教育厅科技计划项目(GJJ13676)

摘　　要：目的:研究中药青葙子中的齐墩果烷型三萜青葙苷A(celosin A,CA)对人肝癌HepG2细胞凋亡的诱导作用及其相关机制。方法:采用E-MEM培养基建立人肝癌HepG2细胞体外培养体系,采用细胞计数试剂盒(,CCK-8)检测细胞活力,噻唑蓝(MTT)法测定乳酸脱氢酶(LDH),免疫比色法检测细胞增殖情况,用4',6-二脒基-2-苯基吲哚(4,6-2 amidine base-2-phenyl indole,DAPI)荧光染色观察细胞凋亡形态学变化,Western blot检测半胱氨酸天冬氨酸蛋白酶(Caspase)-3/7和核转录因子kappa B(NF-κB)蛋白表达水平。结果:用0.22-3.5μmol·L-1CA处理过的HepG2细胞,LDH释放增加,呈剂量和时间依赖性。在CCK-8和5-溴脱氧尿嘧啶核苷(BrdU)测定过程中,CA比标准药物抗HepG2活性更强,随浓度和作用时间的增加,细胞的活性不断下降,对HepG2增殖的抑制作用明显。DAPI染色观察用CA处理过的HepG2细胞浓缩明显,细胞核分散,染色质凝聚。Western blot结果显示用CA处理过的HepG2细胞Caspase-3/7的活性显著增加,NF-κB蛋白表达水平下降。结论:Celosin A具有诱导HepG2细胞凋亡的作用,其机制可能与激活Caspase-3/7和抑制NF-κB蛋白表达有关。Objective： To investigate the apoptosis in human hepatic cancer HepG2 cell strain induced by celosin A（CA） from Celosia Semen and its mechanism. Method： The viability of HepG2 cells was detected by the method cell counting kit-8 （CCK-8）.The activity of lactic dehydrogenase （LDH） was measured by thiazolyl blue（MTT）assay.Immune colorimetry was used to test the proliferation of HepG2 cells.The change of morphoiogy of apoptosis HepG2 cells was observed by 4,6-2 amidine base-2-phenyl indole（DAPI） fluorescence staining.Western blot was used to detect the expressions of Caspase-3/7 and nuclear factor kappa B（NF-κB） proteins of HepG2 cells. Result： After HepG2 cells were treated by CA of different concentration,the leakage of LDH increased in a dosage and time dependent manner.In the process of determination of BrdU and CCK-8,the inhibitory effect of CA to HepG2 cell was stronger than standard drugs,and showed clear inhibitory effect on the HepG2 cell proliferation with the increase of concentration and the acting time.By using the DAPI fluorescence dyes,the HepG2 cell cytoplasm concentrating,nucleolus dispersion and chromatin aglomeration were observed.The Western blot analysis showed that the expression of Caspase-3/7 was significantly enhanced and the expression of NF-κB proteins was down-regulated. Conclusion： Celosin A could induce the apoptosis of HepG2 cells and its mechanism related to activating the activity of Caspase-3/7 and down-regulating the expression of NF-κB protein.

关 键 词：青葙子 青葙苷A HEPG2细胞 凋亡 核因子-ΚB 

分 类 号：R285.5[医药卫生—中药学]