Mfge8基因敲除纯合子小鼠的定向繁殖和鉴定及其与自身免疫疾病相关性分析  

Breeding and identification of Mfge8 knockout homozygous mice and their association with autoimmune diseases

作　　者：杨文强[1] 黄伟[1] 马威[1] 崔天盆[1] YANG Wenqiang;HUANG Wei;MA Wei;CUI Tianpen(Central Laboratory, Wuhan No. 1 Hospital, Wuhan 430022, China)

机构地区：[1]武汉市第一医院中心实验室,武汉430022

出　　处：《中国比较医学杂志》2019年第5期69-75,共7页Chinese Journal of Comparative Medicine

基　　金：国家自然科学基金(81770038);武汉市卫计委2017临床医学科研重点项目(WX17A01);武汉市卫计委2017临床医学科研项目(WX17Z01)

摘　　要：目的繁殖和鉴定Mfge8基因敲除C57BL/6小鼠,获取纯合子(Mfge8^(-/-))小鼠,并比较Mfge8^(-/-)小鼠血清学及组织学改变。方法对幼鼠剪尾抽提基因组DNA,用PCR法和琼脂糖凝胶电泳法鉴定幼鼠基因型;利用TUNEL法分别检测12周龄野生型(WT)和Mfge8^(-/-)小鼠肺组织中凋亡细胞;并且利用免疫荧光法分别检测36周龄WT和Mfge8^(-/-)小鼠血清中ANA和AECA含量。结果构建的基因敲除小鼠已成功繁育保种,并得到纯合基因缺失型小鼠。同时,12周龄Mfge8^(-/-)小鼠肺组织中凋亡细胞数量明显多于WT小鼠;36周龄Mfge8^(-/-)小鼠血清中ANA抗体及AECA抗体阳性,而WT小鼠血清中呈现阴性。结论 C57BL/6背景小鼠敲除Mfge8基因后,更易发生自身免疫疾病。Objective To breed and identify Mfge8 gene knock-out (Mfge8^-/-) C57BL/6 mice, and to examine serological and histological changes in these animals. Methods Genomic DNA was obtained from the tail of young mice and the genotype was determined by PCR and agarose gel electrophoresis. TUNEL assay was used to detect apoptotic cells in the lung tissues of 12-week-old wild type (WT) and Mfge8 ^-/- mice. Levels of anti-nuclear antibodies (ANA) and antiendothelial cell antibodies (AECA) in the serum of 36-week-old WT and Mfge^-/- mice were determined by immunofluorescence. Results Knockout mice have been successfully generated and maintained that are homozygous for Mfge8genedeletion(Mfge8 -/-).The number of apoptotic cells in the lung tissues of 12-week-oldMfge8 -/-micewashigher than in WT mice. In addition, the serum of 36-weeks-old Mfge8 -/- mice, but not WT mice, was positive for ANA and AECA. Conclusions Mfge8 knockout mice with the C57BL/ 6 genetic background demonstrate a greater susceptibility to autoimmune disease.

关 键 词：Mfge8 纯合子 凋亡细胞 ANA 自身免疫疾病 小鼠 

分 类 号：R-33[医药卫生]