Beta2-GP1Affect Platelet Membrane Leads To Platelet Aggregation And Promote The Formation Of DVT

[Objective]DVT pose a threat to life, the mechanism is complex,so we need further research.In our study,we build rats model of DVT,collecting venous blood beofre thrombosis formed, and fully formed; In our clincal research, we collected the venous blood before thrombosis formed, then both extract plasma, quantitative detected of beta2-GP1protein expression; Then we use a variety of bioinformatics analysis technology to explore the specific value of deep vein thrombosis formed in molecular level, Mainly focus on endothelial cells、 platelets and inflammatory cells, for further analyzes the connection of platelet and thrombosis at the molecular level, Thus we study the connection between platelets and DVT at molecular level more specific and effective.[Methods]Part1In the rat model of DVT plasma, detecting beta2-GP1protein expression. Building the model and grouping:70female SD rats, any age, were randomly divided into three groupsNormal control group (10rats):Normal breeding, without any intervention. The control group (30rats):Abdomen anesthesia, laparotomy, no separation, exposure to blood vessels, closed abdomen, normal breeding.The experimental group (30rats):Abdomen anesthesia, laparotomy,, Separation of inferior vena cava,5-0Ethicon sutured placed along a parallel, close to the inferior vena cava, then use4-0ethicon sutured ligation of the inferior vena cava, Draw again go side by side5-0sparing ethicon, closed abdomen, normal breeding.2Blood collection and pathologic examination:In building models after2h,8h,24h, Randomly chose each10rats from control group, experimental group.After anesthesia, collecting blood3-8ml via the inferior vena cava ligation place above (0.5cm), place in the sodium citrates, centrifugal at3000r/min for15minutes, separated plasma, freezing in-80℃fridge;3Mixed respectively at different groups’plasma samples, ELISA to detect beta2-GP1expression4Statistical analysis:Spss17.0software analysis data, Measurement data with X+S, difference with single factor analysis of variance between groups, P<0.05was statistically significant.Part2β2-GP1in DVT patients’plasma 1Thrombosis diagnosis and grouping:according to 《NICE guideline》 and 《Methodology for the Developmentof Antithrombotic Therapy and Prevention of Thrombosis Guidelines》, to determine the criteria for the diagnosis of DVT. Selection in August2013-December2013, in the first affiliated hospital of kunming medical university orthopaedic, vascular surgery, dry treatment of30cases of hospitalized patients with DVT. Selecting orthopaedic for thrombosis did not form a group,30patients with spinal cord injury, ischemic necrosis of femoral head and knee osseous arthritis, hip fracture; Relief to spine fracture fixation, total hip replacement, a hip replacement, hip and knee arthroplasty, open reduction and internal fixation of fracture as the main treatment; Selected injury patients are more likely to form thrombus, surgical treatment are closely related and thrombosis, action inconvenience, these patients in long-term and high risk factors of thrombosis is used. At the same time, select15healthy volunteers. Normal control group:Randomly selected10healthy volunteers Thrombosis group:In30cases of thrombosis patients from15cases Thrombosis unformed group:Dynamic observation2weeks without thrombosis30cases,15cases were selected2Blood samples collected:Collect each upper limb peripheral venous blood,5ml/time, place in the sodium citrates, centrifugal at3000r/min for15minutes, separated plasma, freezing in-80℃fridge 3. ELISA to detect beta2-GP1expression:Mixed respectively at different time、different groups’plasma samples, ELISA to detect beta2-GP1expression4Statistical analysis:Spss17.0software analysis data, Measurement data with X+S, difference with single factor analysis of variance between groups, P<0.05was statistically significant.Part3bioinformatics analysis technology to explore the specific value of deep vein thrombosis formed in molecular level1application Kegg pathway database, analysis, induction, summarizes the beta2-GPI and platelet aggregation in how to affect the coagulation of dynamic balance.[result]Part1In the rat model of DVT plasma, detecting beta2-GPI protein expression.2After modeling, In the corresponding time points to take plasma, normal controlβ2-GP1’OD:0.43±0.24, the control group:0.44±0.12, thrombosis unformed group:0.58±0.14, early time of Thrombosis formed:0.74±0.16, peaking time of Thrombosis formed:0.80±0.10、β2-GP1gradually rise in thrombosis unformed group,early time of Thrombosis formed, peaking time of Thrombosis formed, No significant change in the control group.3β2-GP1’s OD in different groups:The control group/normal control group, P=0.112, No statistical differences; thrombosis unformed group/normal control group, P=0.033, thrombosis unformed group/The control group, P=0.037, There are Statistical differences early time of Thrombosis formed/normal control group,P=0.000, early time of Thrombosis formed/The control group, P=0.006, here are Statistical differences peaking time of Thrombosis formed/normal control, P=0.000, peaking time of Thrombosis formed/The control group, P=0.001, there are Statistical differencesPart2β2-GP1in DVT patients’plasma1In the corresponding time points to take plasma, normal controiβ2-GP1’OD:0.22±0.06, thrombosis unformed group:0.23±0.05, Thrombosis group:0.27±0.07。2β2-GP1’s OD in different groups: Thrombosis group/normal control group, P=0.018, there are Statistical differences; thrombosis unformed group//normal control group, P=0.318, No statistical differences; Thrombosis group/thrombosis unformed group, P=0.048,, there are Statistical differences; Part3Application of bioinformatics analysis technology, platelet and DVT at the molecular level1beta2-GPI apoER2, GPIb alpha receptors by platelet membrane surface, AKt and Enos downstream genes, influence PGG2and PGH2metabolism, TXA2release, TXA2role in platelet TBXA2R receptor, activation of PLC beta, reduce the content of CAMP and activity, make the platelet membrane glycoproteins (GP) Ⅱb/III a site exposure, caused by platelet aggregation and activation of platelet adsorption III clotting factors and start the extrinsic coagulation pathway, causes the plasma prothrombin into thrombin, clotting enzyme catalyzed fibrinogen into filamentous fibrin, promote thrombosis.[Conclusion]Beta2-GPI in rats, DVT expression level in plasma, mainly through induced platelet membrane surface apoER2and GPIb alpha receptors, membrane phospholipids TXA2release, effect on the platelet membrane GPIIb/III a, enhanced platelet aggregation, to further increase the platelet membrane adsorption of clotting factor III, start the extrinsic coagulation pathway, promote deep vein thrombosis.