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Anti-thrombosis effect of LRRFIP1 shRNA lentivirus in a mouse model of deep vein thrombosis
Authors:Xiang Yin  Siyu WuZiming Wang  Yu WangQuanyin Du  Aimin Wang
Affiliation:Department of orthopaedics, Institute of surgery research, Daping Hospital, Third Military Medical University, Chongqing 400042, PR China
Abstract:

Introduction

Deep vein thrombosis (DVT) is one of the common complications of orthopedic surgery. Low molecular weight heparin (LMWH) is a usually used agent for DVT, but it would increase the risk of bleeding. LRRFIP1 has been shown to play an important role in the formation of thrombosis. Therefore, we investigated the effect of LRRFIP1 shRNA lentivirus on DVT in mice.

Materials and Methods

Lentiviral Vectors carrying LRRFIP1 shRNA were constructed and transfected into cultured mouse bone marrow cells (BMCs). Male ICR mice were irradiated with a single dose of 9.5 Gy and then were injected with different agents through the tail vein. Stasis venous thrombosis was induced by inferior vena cava (IVC) ligation. Mice were sacrificed on the 1st, 3rd and 7th day post operation and the thrombi were removed, blotted the excess blood on it with filter paper and immediately weighed. P-selectin and d-Dimer were determined by enzyme-linked immunosorbent assay (ELISA).

Results

LRRFIP1 shRNA significantly suppressed the expression of LRRFIP1 in the thrombi. In contrast, low molecular weight heparin (LMWH) and negative shRNA exhibited little effect on the expression of LRRFIP1. LRRFIP1 shRNA, LMWH and negative shRNA inhibited the thrombus formation in vivo significantly. The plasma P-selectin and d-Dimer levels were significantly increased after IVC ligation. LRRFIP1 shRNA significantly decreased the plasma P-selectin and d-Dimer levels. However, LMWH and negative shRNA showed little effects on the levels of plasma P-selectin and d-Dimer.

Conclusion

LRRFIP1 shRNA might represent a promising prevention strategy for DVT.
Keywords:bone marrow cells, BMCs   deep vein thrombosis, DVT   enzyme-linked immunosorbent assay, ELISA   green fluorescent protein, GFP   inferior vena cava, IVC   leucine-rich repeat in Flightless-1 interaction protein 1, LRRFIP1   low molecular weight heparin, LMWH   pulmonary embolism, PE   small interfering RNA, siRNA   total hip arthroplasty, THA   total knee arthroplasty, TKA
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