Expression Of C-Kit+ Cells And Sca-1+ Cells In Coronary Atherosclerotic Calcification Model Of Rats

BackgroundAtherosclerosis is a kind of chronic disease after vascular endothelial injury.In recent years,the morbidity trend is younger.Atherosclerosis is often accompanied by calcifica-tion,which is one of the signs of atherosclerosis and its inevitable result.Recent views suggest that atherosclerotic calcification is an active process rather than simply passive deposition of calcium crystals.Sca-1 and c-Kit,as markers of stem cells,are widely used in cardiovasc-ular disease research.To explore the changes of Sca-1 and c-Kit expression in atheroscle-rosis with calcification may provide new mentality for the treatment of atherosclerosis with calcification.ObjectiveTo investigate the expression of Sca-1 and c-Kit in coronary atherosclerosis calcifica-tion rats,and to provide a cytological basis for further study of coronary atherosclerosis.Methods1.Thirty-two healthy adult SD rats were randomly divided into two normal control group(n=8)and coronary atherosclerotic calcification model group((n=24).The control group was fed with conventional diet.The model of atherosclerosis calcification rats was established by using intragastric administration of propylthiouracil,feeding with high fat and cholesterol diet and injecting vitamin D3 though intraperitoneal.2.When the seventh week,two rats were slected from the normal group and the model group randomly.The rat hearts were embedded with OCT,then frozen section.Oil red O,alizarin red and HE staining were used to observe the pathological changes of coronary artery in model rats.The model group was formed at the twelfth week.3.Immunohistochemistry technique were used to detect the expression of Sca-1,c-Kit.4.Immunofluorescence technique were used to detect the co-expression of Sca-1and?-SMA.5.Western Blotting technique was used to detect the difference of c-Kit and Sca-1protein expression between model group and normal group.Results1 At 12 weeks,the coronary artery of model group was obviously thickened,intima was obviously proliferated,smooth muscle arranged in disorder,necrotic substance deposition,lipid deposition and calcium nodule precipitation appeared.It is suggested that the model of atherosclerosis calcification is successfully established.2 Immunohistochemistry results showed that the expression of Sca-1~+cells and c-Kit~+cells in intima,media and adventitia of coronary artery in atherosclerotic calcification rat model was significantly higher than that in normal group.3 Immunofluorescence results showed that the expression of Sca-1~+cells and c-Kit~+cells in the coronary artery of atherosclerotic calcification rat model was consistent with the results of immunohistochemistry,co-expression of Sca-1 and?-SMA in some cells.4 Western Blotting results showed tha the protein content of c-Kit and Sca-1 in athero-scler otic calcified rats was higher than that in the normal group.ConclusionThe number of Sca-1~+cells and c-Kit~+cells increased under the condition of coronary atherosclerosis calcification,and the mechanism needs to be studied.