人视黄醇结合蛋白4多克隆抗血清的制备及在妊娠糖尿病中的初步应用  

Preparation of a polyclonal antiserum to human RBP4 and its preliminary application in gestational diabetes mellitus

作　　者：陈利春[1] 鲁云霞[2] 孙玉秀[1] 方向东[1] 叶俊良[1] 陈磊[1] 李朝飞[2] 

机构地区：[1]安徽省妇幼保健院,安徽医科大学妇幼保健临床学院,合肥市230001 [2]安徽医科大学生化教研室,合肥市230032

出　　处：《实用医学杂志》2013年第7期1050-1052,共3页The Journal of Practical Medicine

基　　金：合肥市科技局基金项目(编号:合科[2011]25号)

摘　　要：目的:制备抗人视黄醇结合蛋白4(retinal binding protein4,RBP4)的多克隆抗血清并用于妊娠糖尿病(GDM)血液和胎盘的检测。方法:用自制rhRBP4蛋白免疫新西兰兔制备多克隆抗血清,Western blot检测特异性,ELISA法测定效价,收集20例GDM患者和15例正常孕妇的血液和胎盘,检测RBP4的水平和分布。结果:制备的抗血清效价为1∶128000。与正常孕妇相比,GDM患者血清中RBP4水平明显升高,其分娩胎盘的滋养层细胞质和胞膜中RBP4也显著高表达。结论:制备的多克隆抗血清可初步用于GDM患者血清或组织样本中RBP4的检测,为进一步用于临床检测奠定基础。Objective To prepare the rabbit polyclonal antiserum against retinal binding protein 4 （RBP4） which could be applied to detect blood and placental in gestational diabetic mellitus （GDM）. Methods The polyelonal antiserum to rhRBP4 were prepared by immunizing New Zealand rabbits with self-made protein rhRBP4. The specificity and titer of antiserum were determined by western blot and ELISA respectively. RBP4 levels and distributions in serum and placenta of 20 GDM patients and 15 normal pregnant women were analyzed by western blot and immunohistochemistry respectively. Results The titer of polyclonal antiserum to rhRBP4 was about 1：128 000. RBP4 serum levels of GDM patients were significantly higher than normal pregnant women, mainly distributed in the cytoplasm and membrane of trophocyte. Conclusion The polyclonal antiserum were successfully prepared and could be preliminary used to detect RBP4 in serum and tissue samples of GDM patients, which laid as a foundation for further clinical detection.

关 键 词：糖尿病 妊娠 重组人视黄醇结合蛋白4 多克隆抗血清 血清 胎盘 

分 类 号：R714.25[医药卫生—妇产科学]