Synthesis And Metabolism Of Sulfatide In Atherosclerosis During High Fat Diet

Objective: Previously,we have known that the change of serum sulfatide content is related to the occurrence and development of atherosclerosis,and by detecting the content of sulfatide in various organs,we found that different organs have different sulfatide content and different composition.In this experiment,we established an animal model of atherosclerosis.On the basis of previous experimental conclusions,we detected the changes of sulfatide synthetase and Catabolic enzyme levels in multiple organs to find out the possible main source of serum sulfatide.The aim is to clarify the synthesis and metabolism of serum sulfatide and achieve the purpose of regulating the content of sulfatide in serum,or disease diagnosis and treatment of atherosclerosis series provides a new thought and method.Methods:A model of atherosclerosis was established in 10 the healthy male New Zealand white-eared rabbits fed a high-fat diet.Firstly,in the normal mode,rabbits were fed with normal diet for 7 days and randomly divided into two groups,namely,5 rabbits in the high-fat model group and 5 rabbits in the normal control group.The model group was fed with the same amount of high-fat feed every day,and the control group was fed with the same amount of ordinary feed every day.The diet of each rabbit was controlled at 135g/ day,ensured it could drink water freely.Secondly,the body weight,blood lipid levels and serum thiobrenoside levels of the two groups were measured before feeding,4 weeks,8 weeks,12 weeks and 16 weeks,respectively.The blood should be fasted for 12 h before each blood collection.Finally,after 16 weeks of high-fat diet feeding,the animals were killed,and the liver,kidney and small intestine tissues of the rabbits were collected and pathological sections were made to observe the accumulation of sulfatide in each tissue.At the same time,the expressions of CST and ARSA in liver,kidney and small intestine were determined byreal-time fluorescence quantitative PCR and Western blot analysis,and the synthesis and catabolic enzyme of sulfatide were studied by their changes in levels.It is expected to regulate the activity of CST and ARSA to control the metabolism of thiobrenoside,so as to affect the content of serum sulfatide,and provide a new idea for the treatment of arteriosclerosis and other cardiovascular diseases.Results:1.Hyperlipidemia and atherosclerosis models were successfully established by feeding rabbits with high-fat diet for 16 weeks.Compared with the control group,the serum levels of TC,LDL-C and HDL-C in model group were significantly increased(P<0.05).2.Hematoxylin and eosin(HE)staining of aorta showed that compared with the control group,the aorta intima of the model group was significantly thickened,rich in foam cells and significantly narrowed in the vascular lumen.3.Oil red O fat staining of frozen sections showed that,comparedwith the control group,the liver and kidney samples of the model group had excessive accumulation of adipose tissue,and the small intestine samples had no accumulation of adipose tissue;4.WB analysis and quantitative real-time PCR showed that compared with the control group,the expression of CST in liver and kidney tissues of model group was significantly higher than that of normal group(P<0.05),while the expression of ARSA in the two groups had no significant difference.5.Serum sulfatide level in model group was significantly higher than that in control group(P<0.05).Conclusion:In the atherosclerotic model of rabbits,gradually high-fat diet feeding until the final success of the model,through the experimental results,we found that the change of serum sulfatide content can better reflect the occurrence and development of atherosclerosis than the conventional detection index,therefore,it may replace the current detection factor to become a new detection index of atherosclerotic diseases in the future;at the same time,the levels of CST and ARSA in liver,kidney and small intestine were compared between the two groups,We found that theoverexpression of CST in liver and kidney is the main reason for the increase of serum sulfatide,which urges us to consider the molecular mechanism of precise regulation of CST gene from the perspective of metabolism,which is an important point for the treatment of cardiovasculardisease in the future.