The Research Of Funtion Of TOLL-like Receptor 3 In Atherosclerosis

Objective:To investigate therelationship between TLR3 expressionand IFN-β expression in peripheral blood monouclear cells in patients with lower limbs ASO;To investigate therelationship between TLR3 expressionand IFN-β expression in both plasma and aortain AS mice model,And the relationship between dendrtic cells’ maturation and Toll like recepter 3 in AS mice model.Methods:1.Detection of positive rate of TLR3 and IFN-β in humen’s peripheral blood monouclear cellsSpecimens were divided into two groups: patients with lower limbs ASO group（n=20）and health group（n=20）.Specimens were treated by EDTA for anti-coagulation,and then treated by red blood cell lysis buffer.Abandon upper liquid after centrifugal operation in order to get white blood cells.Then specimens were treated by rabbit anti-human CD14 FITC labelled antibody,rabbit anti-human CD283（TLR3）PElabelled antibody,rabbit anti-human IFN-β APClabelled antibody and Ig G1 PElabelled antibody.After that,positive rate of TLR3 and IFN-β in humen’s peripheral blood monouclear cells were detected by flow cytometry method.2.ApoE^-/-mice were fed in high fat diet to establish AS mice model5 week old ApoE^-/-mice housed under SPF conditionswere fed in high fat diet（n=24;female=12,male=12）,Mice were splited into two groups randomly: mice in experimental group（n=14;female=7,male=7）and mice in control group（n=10;female=5,male=5）.3.Poly（I:C）activate TLR3 in AS mice modelMice in experimental group were injected with poly（I:C）intraperitoneally（ip）,Mice in control group were injected with normal saline ip.After 10 weeks,the mice were sacrificed.Whole blood was drawn by jugular vein puncture.After the blood was added to DETA,the supernatant was kept in liquid nitrogen after centrifugation.And the arotas were removed and preserved in liquid nitrogen.4.Detection of levels of blood fat and glucose in plasma,and levels of TLR3,IRF3,IFN-β and IL-1β in both plasma and aortaLevels of LDL-C,HDL-C,TG,TC and Glu in plasmawere detected by biochemical analyzer.Levels of TLR3,IRF-3,IFN-β and IL-1βin plasma and arota were detected by ELISA method.5.Detection ofquantityand distribution of mature DCs in AS mice modelQuantityand distribution of mature DCs in aorta in both groups’ mice were detected by immunohistochemistry method.Results:1.Compared with control group,TLR3 expression positive rates in peripheral blood mononuclear cells in patients with lower limbs ASO were significantly higher（P<0.001）.2.Compared with control group,IFN-β expression positive rates in peripheral blood mononuclear cells in patients with lower limbs ASO were significantly higher（P<0.01）.3.Compared with ApoE^-/-mice in control group,levels of LDL-C,HDL-C,TG and TCin serumin experimental group were shown had no significant difference（All P>0.05）.But,level of Glu in serumin experimental group were shown significantly descend（P<0.001）.4.Compared with ApoE^-/-mice in control group,levels of TLR3,IRF3 and IFN-β both in plasma and in aorta in Poly（I:C）i.p.group were shown significantly higher（All P<0.001）;5.Compared with ApoE^-/-mice in control group,levels of IL-1β both in plasma and in aorta in Poly（I:C）i.p.group were shown significantly lower（P<0.001）.6.Mature DCs were shown in aorta tissue in ApoE^-/-micein both Poly（I:C）i.p.groupand normal saline i.p.group,and maturedendritic cells mainly distributed in intimal plaque or around the arterial outer membrane surrounding area.But,compared with control group,the quantity of MHC-II+,CD11c+ and CD86+ dendritic cells were significantly higher.Conclusion:1.Disease of lower limbs ASO can cause up-regulation of TLR3 secretion in peripheral blood mononuclear cells;2.Disease of lower limbs ASO can cause up-regulation of IFN-βsecretion in peripheral blood mononuclear cells;3.Poly（I:C）can lead level of Gluin serumin As mice model significantly descend.But,cannot influencelevels of LDL-C,HDL-C,TG and TC in serumin AS mice model;4.Poly（I:C）can lead levels of TLR3,IRF3,IFN-βin bothplasma and aortain AS mice model significantly ascend,and lead level of IL-1βin bothplasma and aortain AS mice model significantly descend;5.Poly（I:C）can promote the progress of DCs’ maturation in AS mice model.