牛乳骨桥蛋白诱导破骨细胞γ-H2AX焦点的形成  

Formation of γ-H2AX foci in osteoclasts induced by bovine osteopontin

作　　者：马雪莹[1,2] 杨巍[1,2] 刘宁[1,2,3] 

机构地区：[1]乳品科学教育部重点实验室,哈尔滨150030 [2]东北农业大学食品学院,哈尔滨150030 [3]国家乳业工程技术研究中心,哈尔滨150086

出　　处：《东北农业大学学报》2012年第2期34-38,共5页Journal of Northeast Agricultural University

基　　金：黑龙江省教育厅创新团队基金(2010td11);东北农业大学创新团队基金(CXT007-3-1)

摘　　要：文章探讨了牛乳骨桥蛋白(bovine Osteopontin,bOPN)是否可引起大鼠破骨细胞(Osteoclasts,OC)γ-H2AX焦点的形成的可能性。采用全骨髓培养法培养破骨细胞,用bOPN处理破骨细胞,MTT比色法测定bOPN对破骨细胞的增殖影响,应用免疫荧光试验检测细胞核内DNA双链断裂(γ-H2AX焦点)的形成。在细胞增殖实验中,bOPN 100μg.mL-1浓度组在24 h较对照组差异显著(P<0.05);100μg.mL-1浓度组在72 h较对照组差异极显著(P<0.01);在免疫荧光试验中,1、10、100和1 000μg.mL-1处理组都可以使OC形成焦点,尤其1 000μg.mL-1处理组时数量虽不多,强度却明显增加。总之bOPN可使H2AX磷酸化,进而诱导γ-H2AX焦点的形成。This paper explored the possibility of bovine osteopontin （bOPN） inducing the formation of γ-H2AX foci in osteoclasts （OC）. Large quantity of high-purity osteoclasts had been cultivated successfully by whole bone marrow culture method. Proliferation of osteoclast was assessed by MTT colorimetry, DNA double strand breaks （DSBs） were determined by immunofluorescecence assay. In cell proliferation experiment, when compared with control group, the osteoclasts proliferation of bOPN 100 μg·mL-1group was notable after 24 h （P〈0.05）, and bOPN 100μg·mL-1 group had more remarkable proliferation after 72 h （P〈 0.01）; in immunofluorescecence assay, 1, 10, 100 and 1 000 μg·mL-1 group can make the formation of γ-H2AX foci in OC, and 1 000 pg.mL＂ group was more intensive though the quantity is less. Overall, bOPN can induce γ-H2AX foci formation by making H2AX phosphowlation.

关 键 词：牛乳骨桥蛋白 破骨细胞 γ-H2AX焦点 荧光检测 

分 类 号：TS201.21[轻工技术与工程—食品科学]