羟基磷灰石纳米颗粒诱导巨噬细胞凋亡及其与HSP70相互关系的研究  

Reseach of hydroxyapatite nanoparticles inducing apoptosis of macrophage and its relation to Hsp70

作　　者：徐东菁[1] 张稷 丁婷婷[1] 孙皎[1] 

机构地区：[1]上海交通大学附属第九人民医院上海生物材料研究与测试中心,上海200023 [2]无锡第五人民医院,江苏无锡214073

出　　处：《功能材料》2007年第9期1533-1536,共4页Journal of Functional Materials

基　　金：国家自然科学基金资助项目(30470479);上海市科委资助项目(0452nm043);上海市重点(特色学科)资助项目(T0202)

摘　　要：研究羟基磷灰石纳米颗粒(HAp nanoparti-cles)引起巨噬细胞凋亡,及其与热休克蛋白Hsp70的相关性。THP-1单核细胞株经PMA诱导成为巨噬细胞,分别加入0、20、100μg/ml HAp纳米颗粒,培养24h后经PI和AnexinV染色,在荧光显微镜下观察,用流式细胞仪检测细胞的凋亡情况,应用western blot方法检测Hsp70蛋白分子表达的改变。显微镜观察显示THP-1单核细胞株能被诱导成巨噬细胞。荧光显微镜显示20、100μg/ml HAp纳米颗粒组凋亡的阳性信号明显强于0μg/ml对照组;流式细胞仪检测结果也证实HAp纳米颗粒组比对照组凋亡率明显增高(P1<0.0001,P2<0.0001),且凋亡的程度呈浓度依赖性(P3=0.025)。Western Blot检测显示20、100μg/ml HAp纳米颗粒组较对照组Hsp70蛋白表达增加,并且随颗粒浓度增高而表达增加。To explore whether HAp nanoparticles induce apoptosis of macrophages and its correlation to Hsp70. Induce THP-1 cell lines to become macrophage with PMA. Co-culture the macrophages with 0,20,100μg/ml HAp nanoparticles in medium for 24h. Collect the macrophages, and dye with PI and AnexinV, then observe and detect the rate of apoptosis of the macrophages; determine the expression level of Hsp70 by Western Blot. THP-1 cell lines can be induced to macrophages. Detection for apoptosis shows that., observing through fluorescence microscope, positive apoptosis signal of 20,100μg/ml HAp particles groups are stronger than that of control group; detecting by flow cytometry（ FCM ）, apoptosis rate of 20,100μg/ml HAp particles groups are also higher than that of control group （P1〈0.0001 ,P2〈0.0001）, moreover, the rate depends on the concentration of HAp nanoparticles （P3 = 0. 025）. Detection by Western Blot result shows the expression of Hsp70 of 20, 100μ/ml HAp particles groups increase comparing to control group, and the level also depends on concentrations of HAp nanoparticles.

关 键 词：羟基磷灰石 纳米颗粒 凋亡 热敏蛋白70 

分 类 号：R318.08[医药卫生—生物医学工程]