Study On The Mechanism Of The Effect Of Foodborne Choline On The Process Of Atherosclerosis

Objective:To investigate the effect of high choline diet on atherosclerosis in male C57BL/6J mice and its mechanism.Research methods: 1.After 1 week of adaptive feeding,24 male C57BL/6J mice were randomly divided into 3 groups: control group(n = 8),choline group(n = 8),choline group(n = 8)and TMAO group(n = 8).TMAO(3g/L)was added to drinking water.The mice in each group were fed normal feed,the drinking water was changed every other day,and the body weight of the mice was recorded every two weeks.after drug intervention for 12 weeks,the mice in the three groups began to fast for 12 hours at 7: 00 pm.Starting from 7 o’clock the next day,after anesthesia with 10% chloral hydrate(350mg/kg),all mice were killed by taking blood from the eyes,and peripheral blood was taken to dissect the liver tissue and thoracic aorta,some of which were fixed with paraformaldehyde and some frozen to be tested.2.After the mice were killed,the whole blood was centrifuged at 3000 rpm for about 15 minutes,and the upper blood samples were taken to detect the concentrations of serum lipids,total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL)and high density lipoprotein cholesterol(HDL)by automatic biochemical analyzer.The thoracic aorta of mice was carefully dissected under anatomical microscope,and the pathological changes of thoracic aorta were observed by hematoxylin-eosin(HE)staining,the expression of scavenger receptor CD36 and SRA1 in thoracic aorta wall was detected by immunohistochemistry(IHC),and the expression of FMO3 in liver was determined by Western blot.result:1.Regularly record the weight of mice,and it was found that the weight of the three groups of mice was basically the same at the beginning of the experiment,with no statistically significant difference(P>0.05).As the experiment progressed,compared to the control group,C57BL/6J mice in the TMAO and choline groups showed significant weight gain,and statistical differences began to appear at the sixth week(P<0.05).At the end of the experiment,the weight of the mice also increased significantly(P<0.05).Compared with the TMAO group,there was no significant statistical difference(P>0.05)in the weight change of experimental mice in the cholinegroup.2.The results of the fully automated biochemical analyzer showed that there was no statistically significant difference in blood lipid levels between C57BL/6J mice in each group(P>0.05).3.HE staining results showed that compared with the control group,C57 BL /6J mice inthe TMAO group and choline group had thickened arterial intima,disordered arrangement of aortic vascular endothelial cells,severe lipid deposition,and a large number of inflammatory cells and foam cells.Compared with TMAO group,mice in choline group also had atherosclerosis;4.Immunohistochemical staining was used to detect the expression of scavenger receptors CD36 and SR-A1 in the thoracic aorta wall of C57BL/6J mice treated with TMAO and choline compared to the control group(P<0.05);5.Protein immunoblotting was used to detect the expression of FMO3 in the liver of C57BL/6J mice.The results showed that compared with the control group,the expression of FMO3 in the liver of TMAO group and choline group mice was significantly increased(P<0.05).Conclusion: Choline may promote the development of atherosclerosis in C57 BL /6J mice through FMO3 /scavenger receptor CD36 and SR-A1 pathways,and thisprocess does not change the level of serum lipids.