Establishment And Experimental Study Of Deep Venous Thrombosis In SD Rats Model

Objective:By establishing the deep vein thrombosis of SD rats model,we observe the changes of color Doppler ultrasound thrombus thickness/vessel diameter ratio,postoperative/preoperative blood perfusion ratio and thrombus length of Peri Cam PSI blood perfusion imaging system(PSI system for short)in SD rats at different time points after modeling.Serum differential proteins(CD138 and S100A8)in patients after trauma were screened by proteomics technology.The protein expression trend of deep venous thrombosis in rats was observed by immunofluorescence staining.It is predicted that CD138 and S100A8 can be used as biomarkers for detection of thrombosis and recurrence monitoring,which can provide help for early diagnosis of deep venous thrombosis of lower limbs.Methods:A total of 88 SD rats were randomly divided into 24 rats in the control group and 64 rats in the thrombus model group.Rats in control group only underwent laparotomy.In the model group,deep vein thrombosis was induced by incomplete ligation of inferior vena cava and complete ligation of branches of inferior vena cava from renal vein to iliac vein and vascular wall damage with vascular clamp.Ultrasound and PSI system were respectively performed at 3h,12 h,1d,2d,3d,7d,14 d and21d after operation to observe the changes of ultrasonic thrombus thickness / vessel diameter ratio and postoperative / preoperative perfusion ratio at different time points of thrombosis.The serum of patients in clinical practice was collected and divided into 10 patients with post-traumatic deep vein thrombosis,10 patients without post-traumatic deep vein thrombosis and 5 healthy people.The serum samples were analyzed by proteomics technology to screen the proteins with significant differences(CD138 and S100A8)for immunofluorescence staining of animal vascular thrombosis specimens,and the protein expression at different time points was observed.Results:A total of 56 deep venous thrombosis models were successfully produced,with the success rate of 87.5 %(56/64).At 1 day,2 days and3 days after modeling,the thrombus thickness / vessel diameter ratio increased gradually and reached the peak at 3 days after modeling.After 7 days of modeling,the thrombus thickness / vessel diameter ratio gradually decreased,reached the minimum on 21 days,and the thrombus completely disappeared in some rats ’ IVC.There was statistically significant difference in the first seven time points between the two groups of ultrasonic testing values(P < 0.05).At 21 days after operation,there was no significant difference between the two groups of ultrasonic testing values(P >0.05).At 1 day,2 days and 3 days after modeling,the postoperative / preoperative perfusion ratio gradually decreased and reached the lowest value at 3 days after operation.At 7days,14 days and 21 days after modeling,the postoperative /preoperative perfusion ratio increased gradually,and reached the highest at 21 days after operation.The comparison of blood flow meter test values between two groups was consistent with that of ultrasonic test.The larger the thrombus thickness / vessel diameter ratio was,the smaller the postoperative / preoperative perfusion ratio was,and there was a negative correlation between the two detection indexes.The length of thrombus increased first,reached the maximum at 3 days after operation,then decreased,reached the minimum at 21 days or part of the model thrombus disappeared basically.Immunofluorescence results showed that :(1)The expression of CD138 in the animal model group was higher than that in the control group.The expression of CD138 reached the maximum on the third day,and then decreased gradually.(2)The expression level of S100A8 showed that the expression level showed an overall increasing trend 2 days ago,and reached the maximum on the second day.The expression level decreased on the first day,and gradually decreased 2 days later.The expression level at21 days was basically the same as that of the normal control group.Conclusion:The postoperative / preoperative perfusion ratio of blood flow meter can be used as an important reference index for thrombosis,which has a certain value in identifying the progress of thrombosis.By changing the modeling sequence of the traditional deep vein thrombosis’ stenosis model,the lateral branches were exposed and ligated,which was conducive to the success of the model.According to the results of proteomics screening and immunofluorescence of vascular thrombosis in rats,it is predicted that CD138 and S100A8 can be used as a biomarker for detecting thrombosis and recurrence monitoring,which provides help for the early diagnosis of deep venous thrombosis of lower extremities.