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2022, Vol. 26 ›› Issue (14): 2196-2201Establishment and identification of a myeloid-derived growth factor deficiency model in apolipoprotein E knockout miceDing Yan1, 2, Xiang Guangda2, Meng Biying2, Xu Xiaoli2, Chen Yuefu1
Abstract: BACKGROUND: There are few studies on myeloid-derived growth factor (MYDGF) and atherosclerosis and metabolic diseases. Therefore, a mouse model of bone marrow specific MYDGF deficiency with apolipoprotein E (ApoE) knockout is particularly important for studying the relationship between MYDGF and atherosclerotic diseases. OBJECTIVE: To establish and identify the mouse model of bone marrow specific MYDGF deficiency with ApoE knockout. METHODS: ApoEflox/flox mice and bone marrow specific MYDGFflox/fiox mice were purchased from the Shanghai Model Organisms Centre, Inc (Shanghai, China). Five ApoEflox/flox male mice were selected to mate with 10 MYDGFflox/fiox female mice, and 30 F1 progeny mice with genotype of MYDGFflox/+ApoEflox/+ were obtained and mated with each other. Finally 30 mice with genotypes of MYDGFflox/floxApoEflox/flox and 34 ApoEflox/flox progenies were obtained. The MYDGFflox and ApoEflox genotypes were identified by PCR, and the body length and body mass of MYDGFflox/floxApoEflox/flox mice and ApoEflox/flox mice at 2 months were recorded. The expression of MYDGF protein in the mouse bone marrow tissues was detected by western blot assay, and immunofluorescence was used to detect the expression of MYDGF in the mouse bone marrow cells. The plaque area of the thoracic aorta in the mouse was detected by oil red O staining. The study was approved by the Animal Ethic Committee of PLA General Hospital of Central Theater Command. RESULTS AND CONCLUSION: There were 34 ApoEflox/flox mice and 30 MYDGFflox/floxApoEflox/flox mice, with no significant difference in their body length and body mass (both P > 0.05). The relative expression level of MYDGF protein in the bone marrow of MYDGFflox/floxApoEflox/flox mice was significantly lower than that of ApoEflox/flox mice (P < 0.05). The percentage of plaque area in the thoracic aorta of MYDGFflox/floxApoEflox/flox mice was significantly higher than that of ApoEflox/flox mice (P < 0.05). Therefore, MYDGF gene was successfully knocked out in ApoEflox/flox mice, and the homozygous mouse model of bone marrow knockout MYDGF gene in ApoE knockout mice was successfully constructed and identified by genotype and protein tissue levels. To conclude, MYDGF gene deficiency can aggravate atherosclerosis in ApoE knockout mice. Key words:bone marrow cells, MYDGF gene, ApoE gene, MYDGF protein, double gene knockout, mouse |