In Murine Different Degree Atherosclerotic Plaques The Expression Of IFN-γ,IL-4 And Correlation Studies With Serum B7-H3

Atherosclerosis is the main cause of death in western developed countries. Along with the improvement of people's living standard and changes of eating habits in our country, the morbidity and mortality of arteriosclerosis also increased year by year. Atherosclerosis accounts for great majority of coronary heart disease(95%-99%).coronary artery acute stenosis or block caused by atheromatous plaque rupturing, surface damaging or crackle occuring, platelet aggregation and thrombosis in coronary artery is the important reasons for acute coronary syndrome. According to current studies, the pathogenesis of atherosclerosis has a variety of theories,such as inflammation damage responses doctrine, fatty infiltration doctrine, platelet aggregation doctrine and smooth muscle cells clone doctrine,while chronic inflammation theory caused by many factors got more and more attention.More and more researches confirmed, in each period of the atherosclerotic occurrence and development, the immune system and inflammation factors play an important role. Compared with high-fat diet fed immunodeficiency mice and the normal mice, two groups both had serious hypercholesterolemia, and two groups of cholesterol had no difference in the distribution of LDL,but atherosclerosis damage of the aortic arch and the thoracic aorta in immune deficiency mice were more significantly reduce than normal mice. So inflammatory immune adjustment mechanism turns into the present research hot spot, the research on this mechanism will break a new pathway for the treatment of AS.By observing the expression of IFN-y and IL-4 in murine different degree atherosclerotic plaques and the relationship with serum B7-H3,this research discussed the function of Cellular immune involved by IFN-y and humoral immune involved by IL-4 in the occurrence and development of atherosclerotic plaques and the mechanism of these cytokines, and maybe provide new ideas for the treatment of atherosclerosis.Research Methods:51 healthy male Wistar rats (weight 220+10g)were randomly divided into normal control group (11) and experimental group (40), and the latter were further divided into High-fat diet group (3w group; 10), Mild atherosclerosis group (4w group; 13), Obviously arteriosclerosis group (5w group;17). normal control group had been raised basic feed and ordinary water for 5 weeks. experimental group had been raised basic feed and ordinary water for 1 week,then were given promoting arteriosclerosis feed and ordinary water.When given promoting arteriosclerosis feed,each rat was given 40 million u/kg weight vitamin D3 by intraperitoneal injection for 3 consecutive days. Promoting arteriosclerosis feed consists of basic feed,2% cholesterol, 0.5% bile acid sodium,0.2% propylthiouracil, vitamin D3 powder (1.25 x 106u/kg feed),3% lard.Sample processing:Rats had been fasting for 12 hours, Measured weight,Treated the above rats with ether inhaled anesthesia, collected some blood using the eyeball-taken-off-method, put the rats death in the method of cervical dislocated, open chest and took artery in its full length, get 1cm artery tissue undernthe aortic arch about 1cm, then immediately fixed the artery tissue with 10% formaldehyde solution for 24 hours, dehydrated, transparencied, leach waxed, embeded for HE Dyeing and immunohistochemi-cal test. Analysed IFN-y and IL-4 expression in vascular tissue with semi-quantitative immunohistochemical method,measured lipid levels,and detected murine serum B7-H3 level with ELISA. Experimental data according to SPSS 16.0 statistical software, proceeds Variance analysis, Pairwise comparison occured with Student-Newman-Keuls among each mean,showed in x±s, P<0.05 indicated statistical significance.Result:1. Weight level:compared with before the experiment, normal control group, high-fat diet group (3 weeks group), mild atherosclerosis group (4 weeks group) and obvious arteriosclerosis group (5 weeks group) had significantly higher weight (P<0.01); After the experiment, there were no obvious difference in each group. Lipid levels:In the experimental group the serum total cholesterol (TC), triglycerides (TG), low density lipoprotein cholesterol (LDL-C) level were increased compared with normal controls (P< 0.01), whereas high-density lipoprotein cholesterol (HDL-c) level was reduced significantly compared with normal control group (P<0.01), as experimental groups fed prolonged, the level of total cholesterol (TC), triglycerides (TG), low density lipoprotein cholesterol (LDL-C) gradually increased (P<0.01), but there was no obvious difference in high-density lipoprotein cholesterol (p>0.05).2.HE Dyeing:Compared with normal control group, in high-fat diet group (3 weeks group),a few small patches in vascular wall are convex to the lumen, and smooth muscle cells in tunica media arrange less regularly, nuclei is like long pole. In mild atherosclerosis group (4 weeks group),several small patches in vascular wall are convex to the lumen, smooth muscle cells in tunica media slightly proliferated,arranging less regularly. In obvious arteriosclerosis group (5 weeks group), vascular wall endangium has big patches centripetally, smooth muscle cells in tunica media obviously proliferated, elastic plates were damaged, lipid drops were melt, and there were foam cells, and deposited purple granular calcium salt.3. Immunohistochemical results:In normal control group, IFN-γand IL-4 of vessel wall endangium and tunica media both expressed a little, which were brown granular;in high-fat diet group (3 weeks group), IFN-γand IL-4 of vessel wall endangium and tunica media expressed more,which were brown granular; in mild atherosclerosis group (4 weeks group),IFN-γand IL-4 of vessel wall endangium and tunica media expressed significantly more,which were brown or dark brown granular and small flake, especially in endometrial plaques; in obviously arteriosclerosis group (5 weeks group) IFN-γand IL-4 of vessel wall endangium and tunica media expressed significantly more,which were brown or dark brown granular and small flake.4.Semi-quantitative immunohistochemical analysis:As the extent of atherosclerosis deteriorated gradually, the expression of IFN-y and IL-4 increased,and the four groups had significantly difference (P<0.001).5.serum B7-H3 level:Compared with normal controls, the B7-H3 expression in high-fat diet group (3 weeks group), mild atherosclerosis group (4 weeks group), obvious arteriosclerosis group (5 weeks group) reduced,but had no statistically differences (P>0.05),and had no difference in experimental groups (P>0.05).Conclusion:1.Cellular immune involved by IFN-γand humoral immune involved by IL-4 were both involved in the formation of atherosclerosis, and could promote arteriosclerosis.2. The formation of the atherosclerosis involved by IFN-y and IL-4 may have no correlation with costimulating molecules B7 H3.