镰形棘豆对SMMC-7721细胞线粒体膜电位和凋亡相关蛋白表达的影响  

Effect of Tibetan Medicine Oxytropis Falcata on Mitochondrial Transmembrane Potential and Expression of Apoptosis Related Proteins in SMMC-7721

作　　者：陈晓红[1,2] 程海清[1] 邓易 杨光明[1] 潘扬[1,3] 

机构地区：[1]南京中医药大学江苏省中药炮制重点实验室国家中医药管理局中药炮制标准重点研究室国家教育部中药炮制规范化及标准化工程研究中心,江苏南京210023 [2]青海大学医学院,青海西宁810001 [3]南京中医药大学药用菌与中药生物技术研究所,江苏南京210023

出　　处：《南京中医药大学学报》2017年第1期54-58,共5页Journal of Nanjing University of Traditional Chinese Medicine

基　　金：国家自然科学基金(81473420;30902012);江苏高校优势学科建设工程资助项目(PAPD);江苏高校品牌专业建设工程资助项目(PPZY2015A070)

摘　　要：目的研究藏药镰形棘豆对SMMC-7721人肝癌细胞活性、线粒体膜电位,细胞色素C(Cyt C)及Caspase-3蛋白表达的影响与细胞凋亡的关系。方法以镰形棘豆总黄酮0.05、0.075、0.1g/L处理肝癌细胞SMMC-7721,24h后观察细胞的生长状况;镰形棘豆总黄酮作用于SMMC-7721细胞后,流式细胞仪检测细胞总体凋亡率和线粒体膜电位,Western blot检测对凋亡相关蛋白Cyt C及Caspase-3表达的影响。结果结果表明,镰形棘豆总黄酮抑制肝癌细胞SMMC-7721的生长,呈浓度依赖关系;镰形棘豆作用后可观察到凋亡细胞的比例升高;可导致细胞的线粒体跨膜电位明显降低,与药物浓度呈负相关。中、高剂量组的细胞中Cyt C及Caspase-3蛋白表达明显升高(P<0.01)。结论镰形棘豆总黄酮诱导SMMC-7721细胞的凋亡可能是通过线粒体途径,并且与影响Cyt C及Caspase-3的表达有关。为镰形棘豆抗肿瘤的作用机制的实验和临床研究提供依据。OBJECTIVE To explore the influence of Oxytropis falcata on proliferation, mitochondrial membrane potential and expression of cytochrome C （Cyt C） and Caspase-3 proteins in SMMC-7721 cells. METHODS SMMC-7721 cells were treated with TFOF for 24 h. Vitality of SMMC-7721 cells was tested by MTT assay. Flow cytometric measurement was used for investigating the effect of TFOF on induction of apoptosis in SMMC-7721 cells and evaluating the effect on mitochondrial membrane potential in cells. Western blot was applied to investigate changes of Cyt C and Caspase-3 expression in cells. RE- SULTS The results showed that TFOF exhibited antiproliferative activity on SMMC-7721 cells in a dose-dependent manner. Flow eytometric analysis demonstrated that TFOF caused a dose-dependent apoptosis of SMMC-7721 cells. After being treated with TFOF of 0.05, 0.075, 0.1 g/L, mitochondrial membrane potential levels of SMMC-7721 cells were lower than control group significantly. Western blot showed that Cyt C and Caspase-3 expression of SMMC-7721 cells was up regulated after TFOF treatment （P〈0.05-0.01）. CONCLUSION The results suggest that O. falcata may be a pro-apoptotic agent for he patic cancer cells through mitochondrial apoptosis pathway and via its modulation of Cyt C and Caspase-3 expression.

关 键 词：镰形棘豆 SMMC-7721 线粒体膜电位 细胞色素C Caspase-3 蛋白表达 

分 类 号：R285.5[医药卫生—中药学]