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Differential expressions of lipid metabolism related genes in the liver of young apoE knockout mice

Ye Hongyan, Yin Miao, Shang Yunju, Dai Xuedong, Zhang Shengqiang, Jing Wen, Du Huiqin, Zhang Liang, Pan Jie

The Key Laboratory of Animal Resistant Biology of Shandong Province, College of Life Sciences, Shandong Normal University.Jinan 250014,Shandong

Abstract

The work was aimed to investigate the differential expressions of lipid metabolism related genes in the early stage of atherosclerosis in the young apolipoprotein E deficient （apoE~(-/-)） mice at different ages with normal chow diet. The genotypes of mice were identified by using multiplex polymerase chain reaction （multi-PCR） analysis. The semi-quantitative reverse transcriptionpolymerase chain reaction （RT-PCR） and real-time quantitative RT-PCR were used to analyze the expressions of lipid metabolism related genes in the liver of apoE4 and age-matched wild type （WT） mice of 14-day old, 1-month old, 2-month old, 3,month old. The serum total cholesterol （TC）, triglycedride （TG）, low-density lipoprotein cholesterol （LDL-C） and high-density lipoprotein cholesterol （HDL-C） contents were assayed using COD-PAP and GPO-PAP methods. The serum apolipoprotein B100 （apoB 100） content was quantitated by immune turbidimetry. The hearts were perfusion-fixed in 4% formaldehyde, infiltrated with 30% gum sucrose for 24 h at 4℃, and embedded in OCT compound. The aortic sinus tissues were serially sectioned at -15℃, stained with Sudan Ⅳ, and counterstained with light green. The results were shown as follows. Compared with that in WT mice, the mRNA levels of apoA Ⅰ and apoA Ⅳ in apoE~(-/-) mice aged from 14-day old to 3-month old changed prominently （P〈0.05）, with apoA Ⅰ up-regulated and apoA Ⅳ down-regulated. At the age of 1 month, the expression of apoB 100 in apoE/ mice was higher than that in WT mice （P〈0.05）. The expression of apoA Ⅴ was up-regulaied （P〈0.05） and there was obvious lipid deposition in the aortic intima in apoE^-/- mice at the age of 2 months. The expressions of fatty acid translocasc （Fat/CD36） and angiopoietin-like protein 3 （Angptl 3） in apoE~(-/-) mice were higher than those in WT mice at the age of 3 months （P〈0.05）, while the expressions of peroxisome proliferator-activated receptor #alpha# （PPAR#alpha#）, liver X receptor #alpha# （LXR#alpha#）, carnitine palmitoyl transferase Ⅰ （CPT Ⅰ） and acyl coenzyme A oxidase Ⅰ （ACOXI） showed no significant changes. The serum TC, TG, LDL-C and HDL-C contents in apoE~(-/-) mice aged from 14-day old to 3-month old were higher than those in age-matched WT mice. apoE~(-/-) mice showed a marked increase in serum apoB100 content, consistent with the trend of serum LDL-C content and apoB100 mRNA content in the liver. The results suggest that the mRNA expressions of apoA Ⅰ, apoA Ⅳ, apoA Ⅴ, apoB 100 and Angptl 3 in apoE~(-/-) mice change significantly compared with those in WT mice, and these genes might be relevant to the complicated lipid metabolism network, and involved in the early stage of atherogenesis.

Key words: apoE knockout mice;Atherosclerosis;lipid metabolism related gene;Liver;RT-PCR

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Citing This Article：

Ye Hongyan, Yin Miao, Shang Yunju, Dai Xuedong, Zhang Shengqiang, Jing Wen, Du Huiqin, Zhang Liang, Pan Jie. Differential expressions of lipid metabolism related genes in the liver of young apoE knockout mice. Acta Physiol Sin 2008; 60 (1): 51-58 (in Chinese with English abstract).