| China's development of health statistical bulletin from Statistical Information Center in 2006 shows that cardiovascular disease and cerebrovascular disease caused by atherosclerosis (AS)poses a serious threat to human health in China. The incidence of the disease and mortality is rising year by year, and the trend has been gradually younger.The formation and development of Atherosclerosis is caused by many factors. Its pathogenesis includes a variety of theories,such as lipid infiltration doctrine, thrombosis doctrine, arterial smooth muscle cell proliferation doctrine and damage responses doctrine.Recently some experts suggest that atherosclerosis is associated with inherent immune and acquired immune of arterial wall injury under the condition of inflammation.Many immune cells plays an important role in progression of atherosclerosis,such as T lymphocytes and B lymphocytes. Recently, someone put forward a new type of CD4+CD25+ regulatory T cells which is a group of T cells playing an important role in immune adjustment.It is important in occurrence and development of atherosclerosis. The number of CD4+CD25+Treg in patients with acute coronary syndromes decreased, and the plaque of atherosclerotic will reduce by transferring this regulatory T cells in animal models. Transforming growth factor-1(TGF-beta1) is one of the representative secretion factors of CD4+ CD25+Treg.In atherosclerosis formation process,it participates in multiple pathways of immune adjustment.On the one hand, it can alleviate damage responses and protect cells,.And on the other hand, it can cause excessive extracellular matrix sedimentary, tissue fibrosis, restenosis, and remodel the blood vessels and cardiac. The expansion of CD4+CD25+Treg in vivo needs TGF-beta 1. It has been found that CD4+CD25+regulatory T cells can be significantly increased in mice by using anti-CD3 antibody,and the organ transplant rejection and autoimmune disease can be treated. However,the study is less whether can it inhibit atherosclerosis by oraling anti-CD3 antibody.By HE staining of aortic arch to abserve atherosclerotic plaque,measuring the lipid levels,analysing TGF-β1 expression in vascular tissue with semi-quantitative immunohistochemical method, and detecting murine serum TGF-β1 level with ELISA,this research studied the relationship between anti-CD3 antibody and TGF-β1, and discussed the possible mechanism of inhibiting atherosclerosis formation by oraling anti-CD3 antibody.Purpose of the research:observing the inhibition of atherosclerosis by oraling anti-CD3 antibody and the relationship with TGF-β1Research Methods:31 healthy male Wistar rats (weight 220±10g), were randomly divided into normal control group (10),atherosclerostic group (12), and the treatment group(9).Measure the weight for every rat.The normal control group is raised basic feed, ordinary water for 5 weeks. The atherosclerostic group is raised basic feed, ordinary water for 1 week,then is given promoting arteriosclerosis feed and ordinary water.When given promoting arteriosclerosis feed,each rat is 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.The treatment group is raised anti-CD3 antibody (25ug/d)for 5 consecutive days.Sample processing: Measure the weight again and measure the lipid levels for every rat.Treat the above rats with ether inhaled anesthesia, collect some blood using the eyeball taken off method, put the rats death in the method of cervical dislocated, open chest and take artery in its full length, get 1cm artery tissue undernthe aortic arch about 1cm, then immediately fix the artery tissue with 10% formaldehyde solution for 24 hours, dehydrate,transparency, leach wax, embed for HE Dyeing and immunohistochemical test. HE staining of aortic arch to abserve atherosclerotic plaque,analyse TGF-β1 expression in vascular tissue with semi-quantitative immunohistochemical method,and detect murine serum TGF-β1 level with ELISA. Experimental data according to SPSS 16.0 Statistical software, proceeds Variance analysis,showed in x±s, P<0.05 indicates statistical significance.Result:1.Weight level:compared with before the experiment, normal control group, arteriosclerostic group and treatment group were significantly higher weight (P< 0.01); After the experiment each group there were no obvious difference in weight.2.Lipid levels:In the arteriosclerostic 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 reduced significantly (P< 0.01); the serum total cholesterol (TC), triglycerides (TG), low density lipoprotein cholesterol (LDL-C) level in treatment group were reduced compared with arteriosclerostic group (P< 0.01), whereas high-density lipoprotein cholesterol (HDL-c) level increased significantly (P< 0.01); there is no difference in the serum total cholesterol (TC), triglycerides (TG), low density lipoprotein cholesterol (LDL-C) level and high-density lipoprotein cholesterol (HDL-c) level between treatment group and normal control group. 3.HE Dyeing:In normal control group, the vascular wall is smooth,the layer of aortic wall is clear,nuclei of endothelial cells slightly convex to flat in the lumen, and smooth muscle cells in tunica media arrange regularly, nuclei like long pole.In arteriosclerostic group, vascular wall endangium have a lot of big patches, the intimal is eccentric thickening,the plaque is filled with foam cells,smooth muscle cells in tunica media have changed morphologically,and many tunica media have calcified or damaged.In treatment group, there are a few small patches in vascular wall convex to the lumen, and smooth muscle cells in tunica media arrange less regularly, nuclei like long pole,the plaque is much less than arteriosclerostic group,and close to normal control group.4. The immunohistochemistry results of TGF-β1 in each group:There are a large number of TGF-β1 expression in smooth muscle cells and cytoplasm cells of normal control group,and the color is deep yellow.The expression in arteriosclerostic group is much less than in normal control group. And the expression in treatment group is much more than in arteriosclerostic group,and more than the normal control group,it is dark brown and flaky colored.5.Semi-quantitative immunohistochemical analysis:use relative gray intensity to present the positive expression of TGF-β1.Compared the TGF-β1 expression among the vascular wall of the three groups:normal control group(129.76±4.621), arteriosclerostic group (171.4±2.708), treatment group (103.08±2.957).So the expression of TGF-β1 in treatment group is the most(P<0.05),and the expression in arteriosclerostic group is the least(P<0.05).6. Murine serum TGF-β1 level with ELISA,expressed by average optical density:Compared the TGF-β1 expression among the three groups:normal control group(0.301±0.023), arteriosclerostic group (0.259±0.040), treatment group (0.368±0.050).So the expression of TGF-β1 in treatment group is the most(P<0.05),and the expression in arteriosclerostic group is the least(P<0.05).Conclusion:1.Small dose of oral anti-CD3 antibody has significant effect of inhabitation of atherosclerotic plaque formation.2.The inhabitation effect of atherosclerosis may be related to upregulation of TGF-β1.3.Anti-CD3 antibody may be become a new therapy of cardiovascular diseases.
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