血清剥夺反应因子(Sdpr)敲除及转基因小鼠的建立与表型分析  

Establishment and phenotypic analysis of Sdpr-knockout mice and transgenic mice

作　　者：李欣悦 石桂英 雷雪裴 黄艺滢 李珂雅 侯丽雅 王凯瑜 汤家鸣 白琳 LI Xinyue;SHI Guiying;LEI Xuepei;HUANG Yiying;LI Keya;HOU Liya;WANG Kaiyu;TANG Jiaming;BAI Lin(NHC Key Laboratory of Human Disease Comparative Medicine,Institute of Laboratory Animal Science,CAMS&PUMC,Beijing Engineering Research Center for Experimental Animal Models of Human Critical Diseases,Beijing 100021,China)

机构地区：[1]国家卫生健康委员会人类疾病比较医学重点实验室,中国医学科学院医学实验动物研究所,北京协和医学院比较医学中心,北京市人类重大疾病实验动物模型工程技术研究中心,北京100021

出　　处：《中国比较医学杂志》2022年第5期7-15,共9页Chinese Journal of Comparative Medicine

基　　金：国家自然基金面上项目(31672374);北京自然基金面上项目(5202024);中央高校基本科研业务费专项资金(3332020051)。

摘　　要：目的建立Sdpr基因敲除小鼠及Sdpr转基因小鼠模型,为Sdpr基因功能的研究提供小鼠模型。方法利用CRISPR/Cas9技术构建Sdpr基因敲除小鼠,利用过表达质粒构建Sdpr转基因小鼠模型,利用PCR鉴定小鼠基因型,利用qRT-PCR和Western blot检测mRNA和蛋白表达情况,获取小鼠组织进行组织病理学分析,利用流式细胞术对小鼠免疫细胞比较分析。结果获得Sdpr基因敲除及转基因小鼠模型。与野生型小鼠相比,基因敲除小鼠Sdpr基因表达降低;组织多处出现炎性灶,肺、脂肪有炎性细胞浸润;肝出现脂肪变性,中央静脉周围可见髓外造血;脾组织红髓出现髓外造血;骨髓中T细胞比例降低,胸腺中CD4^(-)CD8^(-)细胞增多,CD4^(+)CD8^(+)细胞减少。转基因小鼠Sdpr基因表达增加;肝组织轻微脂肪变性;肺泡间隔增宽;脾组织淋巴细胞增生,红髓细胞增多;外周血中B细胞比例增加,M细胞、T细胞比例降低,胸腺CD4^(+)细胞增多,CD4^(+)CD8^(+)细胞减少。结论建立和鉴定了Sdpr敲除和转基因小鼠,可以为研究SDPR蛋白及其相关胞膜窖蛋白在不同组织器官中的作用机理提供模型。Objective To establish Sdpr-gene-knockout and Sdpr transgenic mouse models to provide suitable models for studying Sdpr gene function.Methods Sdpr-knockout mice were constructed by CRISPR/Cas9 technology,while Sdpr transgenic mice were constructed using plasmid overexpression.We identified mouse genotypes by PCR.The mRNA and protein expression levels were determined by qRT-PCR and Western blot.Mouse tissues were obtained for histopathological analysis.We analyzed the immune cells of the knockout mice and transgenic mice by flow cytometry.Results Sdpr-gene-knockout and transgenic mouse models were obtained.Compared with that in the wild-type mice,the Sdpr gene expression level was decreased in the knockout mice.Meanwhile,inflammatory foci appeared in many tissues,such as inflammatory cells infiltrated into the lungs and fat.Hepatic steatosis can be observed,and extramedullary hematopoiesis can be seen around the central hepatic vein.Extramedullary hematopoiesis can be observed in the red pulp of the spleen.The proportion of T cells in bone marrow of the knockout mice decreased,the number of CD4^(-)CD8^(-)T cells in the thymus increased,while CD4^(+)CD8^(+)T cells decreased.In terms of the findings in the transgenic mice,the expression level of the Sdpr gene increased.In addition,their liver cells showed slight fatty degeneration and widening of alveolar septa.Moreover,spleen lymphocytes exhibited hyperplasia and red pulp cells increased.The proportion of B cells in the peripheral blood of the transgenic mice increased while the proportions of myeloid and T cells decreased.Finally,thymus CD4^(+)T cells increased while CD4^(+)CD8^(+)T cells decreased.Conclusions Sdpr-knockout and transgenic mice were successfully established and identified,which can provide models for studying the mechanisms of action of SDPR protein and its related caveolin protein in different tissues and organs.

关 键 词：血清剥夺反应蛋白因子 Sdpr 基因敲除 转基因 小鼠 

分 类 号：R-33[医药卫生]