脂联素基因敲除小鼠建立及鉴定

Establishment and identification of adiponectin gene knockout mice

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摘要目的建立并鉴定脂联素(APN)基因敲除小鼠。方法将雄性APN杂合子小鼠与雌性C57BL/6J小鼠进行交配,获得雄性和雌性APN杂合子小鼠,再将雄性APN杂合子小鼠与雌性APN杂合子小鼠交配,获得子代小鼠。第4周剪取鼠耳组织并提取基因组DNA,采用聚合酶链式反应(PCR)和琼脂糖凝胶电泳进行基因型鉴定。第8周称重,禁食10 h后,测定空腹血糖。随后给予异氟烷麻醉,经眼球采集血液样本,用于血清甘油三酯测定。提取小鼠脂肪组织的蛋白质和RNA,采用Western blot法和荧光定量PCR检测以上组织中的APN的蛋白表达和RNA水平。结果子代小鼠中,野生型(WT)23只(25.56%)、APN杂合子小鼠21只(23.33%)、APN基因敲除(APN KO)小鼠46只(51.11%),成功构建APN KO小鼠模型。基因鉴定结果显示:APN KO小鼠脂肪组织中蛋白质和RNA水平均明显低于WT小鼠,差异均有统计学意义(P<0.05);WT小鼠与APN KO小鼠体质量、空腹血糖、甘油三酯比较,差异均无统计学意义(P>0.05)。结论本研究成功建立APN KO小鼠模型,为深入阐明APN的作用及基因提供了新的实验工具。 Objective To establish and identify adiponectin(APN)gene knockout mice.Methods Male APN heterozygous mice were mated with female C57BL/6J mice to obtain male and female APN heterozygous mice.Then,male APN heterozygous mice were mated with female APN heterozygous mice to obtain offspring mice.In the fourth week,mouse ear tissues were cut and genomic DNA was extracted.Genotype identification was performed using polymerase chain reaction(PCR)and agarose gel electrophoresis.In the 8th week,after fasting for 10 hours,the fasting blood glucose was measured.Subsequently,isoflurane anesthesia was administered,and blood samples were collected from the eyeball for the determination of serum triglycerides.Proteins and RNA were extracted from adipose tissue of mice.The protein expression and RNA level of APN in the above tissues were detected by Western blot and fluorescence quantitative PCR.Results Among the offspring mice,there were 23 wild-type(WT)mice(25.56%),21 APN heterozygous mice(23.33%),and 46 APN gene knockout(APN KO)mice(51.11%),successfully establishing the APN KO mouse model.The results of genetic identification showed that the levels of protein and RNA in the adipose tissue of APN KO mice were significantly lower than those of WT mice,and the differences were statistically significant(P<0.05).There were no statistically significant differences in body weight,fasting blood glucose and triglycerides between WT mice and APN KO mice(P>0.05).Conclusion In this study,the APN KO mouse model was successfully established,providing a new experimental tool for in-depth clarification of the role and genes of APN.

作者张艳吴园张佳琦那堃闫承慧何佳奇 ZHANG Yan;WU Yuan;ZHANG Jia-qi;NA Kun;YAN Cheng-hui;HE Jia-qi(Department of Cardiology,General Hospital of Northern Theater Command,Shenyang 110016,China;Department of Cardiology,Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区北部战区总医院寒地心血管病全国重点实验室心血管内科武汉大学人民医院心内科

出处《临床军医杂志》 2025年第8期795-798,共4页 Clinical Journal of Medical Officers

基金辽宁省科技计划联合计划项目(2024JH2/102600279)。

关键词脂联素基因敲除小鼠基因型 Adiponectin Gene knockout Mouse Genotype

分类号 R-332 [医药卫生]

作者简介第一作者:张艳(1983-),女,辽宁沈阳人,主管技师,硕士;通信作者:何佳奇,E-mail:jiaqihe202@163.com。