【作者】 陈芳；

【导师】 袁琳；

【作者基本信息】 武汉理工大学 ， 生物医学工程， 2008， 硕士

【摘要】 羟基磷灰石纳米粒子对癌细胞的抑制作用是无机纳米粒子的生物学效应,也是生物中心多年来的研究课题。本论文围绕这一课题,在前人研究的基础上,选择“羟基磷灰石纳米粒子对大鼠肝线粒体作用的研究”作为主要研究内容,目的是在细胞器水平上,将纳米粒子与线粒体直接作用,通过检测线粒体琥珀酸脱氢酶活性,探讨纳米粒子对线粒体活性的影响,为进一步研究纳米粒子的抑癌机制提供理论依据。实验通过传统的组织匀浆和差速离心方法来提取和分离大鼠肝细胞线粒体,选择线粒体的标志酶琥珀酸脱氢酶来检测其活性;探讨了线粒体琥珀酸脱氢酶活力检测条件;最后,将不同浓度纳米HAP粒子悬浮液与线粒体直接作用,以没加纳米HAP粒子作为对照,测定线粒体琥珀酸脱氢酶活力,并对数据进行分析,最后得出结果:1.测定得出线粒体琥珀酸脱氢酶的比活性为7.376±0.910u/mg蛋白质,线粒体活性满足实验要求;2.在37℃条件下,两天后线粒体琥珀酸脱氢酶的比活性丧失,而在-70℃条件下,两个月内比活性保持不变;3.纳米HAP粒子随着浓度的增大和作用时间的延长,对线粒体琥珀酸脱氢酶的比活性抑制作用加强,表明HAP纳米粒子对线粒体琥珀酸脱氢酶的比活性抑制有剂量和时间的依赖性。本实验改进了琥珀酸脱氢酶试剂盒的检测方法,使检测快速、方便、成本大大降低;并且在传统细胞水平研究上将纳米粒子直接与线粒体作用。更多还原

【Abstract】 The effect of hydroxyapatite nano-particles on cancer cells is a kind of biological effect of inorganic nano-particles, and it had been studied by Bomedical Materials and Engineering Research Center of Wuhan University of Technology since many years before. On the basis of the former researeh, the author selected "the effect of HAP nano-particles on the mitcochondria of liver rats" as his main research content in the subject. In this study, we have studied HAP nano-particles effect of mitochondria from the perspective of organelles, tested SDH activity of mitochondria, and explored the effect of HAP nano-particles on the mitcochondria. Therefore this study aims to provide the theoretical foundation for further researching inhibitive effect on cancer cell.Liver mitochondria of rat were isolated and extracted by traditional methods, we obtained active mitochondria by the test of SDH which is a key marker of mitochondria; we investigated the condition of SDH test of mitochondria; last, we put different concentrations of HAP nano-particles to directly treat mitochondria with negative control, then conclusions are made:1. The SDH activity of mitochondria was 7.376±0.910 u/mg protain, it fit to our experimental request;2. The SDH activity of mitochondria was higher at 37℃but its activity disappeared after two days, and the same as in two months at -70℃;3. HAP nano-particles inhibit the SDH activity of mitochondria, and became lower and faster compared with negative control depend on time and dose.We bettered the test of mitochondrial activity by SDH kit, which made test more quickly, conveniently and cut down its cost. Moreover we put HAP nano-particles to mitochondria directly, which based on the traditional cell level research.更多还原