摘要
目的：探讨一种简单、有效、快捷的方法来提取颗粒蛋白前体（PGRN）基因敲除（PGRN-/-）小鼠的DNA以及基因的鉴定方法,同时建立鸡Ⅱ型胶原诱导PGRN-/-小鼠的关节炎模型,从形态学、组织学等多角度观察和鉴定鸡Ⅱ型胶原诱导的PGRN /小鼠关节炎模型的特点.方法：将PGRN-/-小鼠与野生C57BL小鼠进行交配,产仔后6周再将杂合小鼠进行交配产仔,剪尾约0.5～1 cm,提取组织DNA,在PCR体系中设计两对引物,一对用来鉴定野生C57BL/6小鼠,另一对用来鉴定PGRN-/-的纯合小鼠.将鸡Ⅱ型胶原与弗氏完全佐剂（CFA）等体积混合后通过尾部皮下注射PGRN-/-小鼠和野生C57小鼠.野生C57BL对照组小鼠和PGRN-/-实验组小鼠分别进行鸡ColⅡ诱导15周后取其病变腿组织进行阿尔新蓝和茜素红染色,腿组织切片进行苏木精-伊红染色,并进行相关统计学分析.结果：成功鉴定出PGRN-/-纯合小鼠;成功建立鸡Ⅱ型胶原诱导的PGRN-/-小鼠关节炎模型.结论：PGRN-/-小鼠对Ⅱ型胶原诱导关节炎的易感性高于野生小鼠.本研究为临床关节炎的研究与治疗提供了很好的动物模型.
Objective：To investigate a simple and efficient DNA extraction method of the PGRN / mice and its identification,with the establishment of the chicken type Ⅱ collagen-induced PGRN-/-mice model of arthritis,model features of chicken type Ⅱ collagen-induced PGRN-/-mice arthritis were observed and identified in morphological,histological and other multiangle observations.Methods：PGRN-/-mice were mated with wild common C57BL mice.Heterozygous mice were mated six weeks after calving,and DNA was extracted from the 0.5-1 cm tails.There were two pairs of primers in the PCR system.One was for identification of wild normal C57BL mice,and the other for PGRN / mice.Chicken Col Ⅱ,mixed in equal volumes with Freund＇s complete adjuvant （CFA）,were subcutaneously injected to the PGRN-/-mice and wild common C57BL mice through the tail.The control group was injected with an equal amount of PBS.After 15 weeks of chicken Col Ⅱ induction,the lesion legs of control groups and treatment groups were stained by Alcian blue and Alizarin red.And the leg tissues were also stained by hematoxylin-eosin.Then statistical analysis was performed.Results：The PGRN-/-mice were successfully identified and PGRN mice arthritis model were successfully established.Conclusion：PGRN-/-mice are more highly susceptible to collagen Ⅱ-induced arthritis comparing with normal C57BL mice.This experiment provides a good animal model for the study and treatment of arthritis.
出处
《解剖学杂志》
CAS
CSCD
北大核心
2014年第4期448-453,共6页
Chinese Journal of Anatomy
基金
国家自然科学基金（81371928,81171697）
教育部新世纪优秀人才支持计划（NCET-12-1090）
人力资源和社会保障部留学回国人员择优资助项目（2011-235）
