长爪沙鼠肝脏基因组DNA甲基化水平检测

doi:10.3969/j.issn.1674-5817.2018.01.008

摘要/Abstract

摘要： 目的用间接ELISA法检测长爪沙鼠肝脏基因组DNA整体甲基化的水平。方法选取新生仔鼠6只(仔鼠组),3月龄鼠12只 [其中青年对照组6只,高脂饮食诱发非酒精性脂肪性肝病(NAFLD)青年模型组6只],8月龄中老龄鼠6只(中老龄),各组均为雄性,按组采集血清(新生鼠除外)用于总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白(HDL)、低密度脂蛋白(LDL)检测,同时采集肝脏标本二份,一份作常规HE染色的形态学观察,一份提基因组DNA,用ELISA法试剂盒检测肝脏基因组DNA整体甲基化的水平[即5-甲基胞嘧啶(5-mc)含量]。结果青年模型组鼠及中老龄鼠均出现高脂血症,血清TC及TG均不同程度显著上升,青年对照组鼠血脂正常。病理学检查显示,青年模型组与中老龄组鼠肝脏出现较为明显的脂肪变性,而青年对照组肝脏无异常,新生仔鼠肝血窦中可见大量成堆分布有单核细胞和红细胞。肝脏基因组DNA整体甲基化水平,青年模型组沙鼠>新生仔鼠>老龄鼠>5对照组鼠。结论长爪沙鼠NAFLD及年龄增加引起的脂肪性变程度与其基因组DNA甲基化水平相关,间接ELISA法检测肝脏基因组DNA整体甲基化水平简便、迅速,成本低,或可以作为评价其表观遗传学的一种方法。

关键词: 长爪沙鼠, 基因组DNA整体甲基化水平, 5-甲基胞嘧啶(5-mC), 间接ELISA法, 非酒精性脂肪性肝病(NAFLD)

Abstract: Objective To detect the genomic DNA methylation level in the liver of Mongolian gerbil by indirect ELISA.Methods Six newborn gerbils (newborn group),12 young gerbils (6 in young control group and 6 in young nonalcoholic fatty liver disease (NAFLD) model group,6 gerbils in the 8-month-old (middle and aged group) were employed,all were male.The serum samples (except newborn group) were collected for detection of the level of total cholesterol(TC),triglycerides(TG),high-density lipoprotein (HDL),low-density lipoprotein(LDL).At the same time,two samples of liver were collected for HE staining and detecting the whole methylation level of liver genomic DNA [ie,5-methylcytosine (5-mC) contentby indirect ELISA kit.Results The levels of serum TC and TG in the young model group and the middle and aged group were significantly difference with those in the young control group (P<0.05).Morphological examination revealed that the young model group and middle and aged groups developed obvious hepatic steatosis.There are a large number of piles of mononuclear cells and piles of erythrocytes in neonatal gerbil liver sinus.Overall methylation test results showed that high fat diet induced model young group gerbils>newborn gerbils>middle and aged gerbils>young control group gerbils.Conclusions The degree of fatty degeneration caused by NAFLD and age in Mongolian gerbil correlated with methylation level of genomic DNA.The method of indirect ELISA for detecting methylation level of liver genome DNA was simple,rapid and economic,which would be developed as a method to evaluate epigenetics of gerbils.

Key words: Mongolian gerbil, Genomic DNA whole methylation level, 5-methylcytosine(5-mC), Indirect ELISA, Nonalcoholic fatty liver disease (NAFLD)

中图分类号:

Q95-33

王志远, 刘月环. 长爪沙鼠肝脏基因组DNA甲基化水平检测[J]. 实验动物与比较医学, 2018, 38(1): 44-47.

WANG Zhi-yuan, LIU Yue-huan. Detection of Whole Methylation Level of Liver Genomic DNA in Mongolian Gerbil Based on Indirect ELISA[J]. Laboratory Animal and Comparative Medicine, 2018, 38(1): 44-47.

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