磷对成骨细胞超微结构及其GJIC和Ⅰ型胶原含量的影响  

Effects of phosphorus on ultramicrostructure,GJIC and content of collagen Ⅰ of osteoblasts in vitro

作　　者：卓丽玲[1,2] 刘宗平[2] 

机构地区：[1]枣庄学院生命科学系,山东枣庄277160 [2]扬州大学兽医学院,江苏扬州225009

出　　处：《中国兽医科学》2011年第8期839-843,共5页Chinese Veterinary Science

基　　金：国家自然科学基金资助项目(30972229;30571364);江苏省高校"青蓝工程"中青年学术带头人培养对象资助项目(2006);枣庄学院博士基金项目(2009);枣庄学院青年项目(2010)

摘　　要：在体外培养成骨细胞(OB)的基础上,添加0、1、2、4mmol/L磷,作用2d后,采用电子显微镜观察OB的超微结构,采用划痕标记染料示踪技术观察细胞间隙连接通讯(GJIC);作用2、5、8d后,用BCA法蛋白定量试剂盒、Goat Anti-rat Collagen TypeⅠ试剂盒分别检测OB内总蛋白及Ⅰ型胶原的含量,并用DⅠ型胶原/D总蛋白表示Ⅰ型胶原的含量。结果显示,作用2d,4mmol/L磷组OB内线粒体减少、肿胀。4mmol/L磷组GJIC的荧光扩散距离减弱,1、2mmol/L磷组基本不变;就Ⅰ型胶原含量而言,除1mmol/L磷组在作用2d抑制其分泌外(P<0.05),在磷作用2、5、8d均促进其分泌(P<0.05)。结果表明,添加1、2、4mmol/L磷能促进OB的分泌Ⅰ型胶原;4mmol/L磷能抑制OB的代谢活性,且能抑制OB间通讯。Osteoblasts（OB） were exposed to 0,1,2 and 4 mmol/L phosphorus in vitro to observe the ultramicrostructure by electronmicroscopy, and to observe the gap junction intercellular communication （GJIC） as well using nick marker dye tracer technology on day 2 post-exposure,and to detect contents of the total protein and collagen type Ⅰ in OB on day 2,5 and 8 post-exposure using BCA method kit and Goat Anti-rat Collagen type Ⅰ kit,respectively. The content of collagen type Ⅰ was expressed by Dcollagen type Ⅰ / Dtotal protein. In result, the mitochondria in 4 mmol/L P group were decreased on 2 day post-exposure. The fluorescence diffusion length of GJIC in 4 mmol/L P group was decreased and in 1 and 2 mmol/L P groups were unchanged on day 2 post-exposure. The contents of collagen type Ⅰ in 1,2 and 4 mmol/L P groups were all enhanced on day 2,5 and 8 post-exposure respectively（P〈0.05） except 1 mmol/L P groups on day 2 post-exposure. In conclusion,P in the all exposure groups enhanced OB to secrete collagen type Ⅰ,and 4 mmol/L P inhibited metabolic activity and intercellular communication of OB.

关 键 词：成骨细胞 磷 超微结构 细胞间隙连接通讯 

分 类 号：S852.21[农业科学—基础兽医学]