Protective Role of Shenfu Injection (SF) in Ischemic Reperfusion Injury of Rat Liver Graft
|School||Chongqing Medical University|
|Keywords||Shenfu Injection Rats Orthotopic liver transplantation Ischemia-reperfusion injury (IRI)|
Objective: This article aims to explore the protective effect of Shenfu Injection (Shenfu injection, SF) on rat liver graft ischemia-reperfusion injury (Ischemic reperfusion injury, IRI) and its specific mechanism from the tissue and cellular level. Methods: 72 healthy male SD (Sprague Dawley) rats, weight 190 to 220g, randomly divided into a control group and SF group (SF group receptor once via the tail vein graft implanted portal vein blood flow recovery soon injection SF 1ml/100g, control group injected with normal saline in the same way), n = 18, using two-cuff technique for orthotopic liver transplantation. Where each portal vein reperfusion at different times, are divided into three groups, respectively completed in liver transplantation 2h, 4h and 6h after collection the receptor of serum and liver tissue, serum tumor necrosis ELISA (enzyme-linked immunosorbent assay) method concentration factor α (tumor necrosis factor-α, TNF-α), immunohistochemistry method for the determination of the nuclear factor κB (nuclear factor kappa B, NF-κB) expression, in situ hybridization method for the determination of cell adhesion molecules (intercellular adhesion molecule-1, ICAM-1) mRNA. Addition, we selected 36 healthy male SD rats, weight 180 to 210g, were randomly divided into a control group and SF group (SF group per well of cell suspension was added dropwise 0.5mlSF control group each hole to give the same amount of saline), each of which reoxidation time is divided into three groups, respectively situ liver perfusion resulting cell suspension into hypoxia - reoxygenation model (simulation in vitro IRI). Reoxygenation 2h, 4h and 6h after taking the cell suspension for testing. Measured by ELISA of ICAM-1 expression, immunocytochemistry method for the determination of the expression of NF-κB. Results: for serum and histological detection rats after liver transplantation is completed, 2h, 4h, 6h, SF group concentration of TNF-α (576 ± 60pg/ml, 518 ± 52pg/ml and 421 ± 35pg / ml), compared to the concentration of the control group (831 ± 65pg/ml and 715 ± 69pg/ml and 598 ± 52pg/ml) was significantly lower (P lt; 0.05); the SF group organization of NF-κB and the expression of ICAM-1mRNA were significantly weaker than the control group. Reoxygenation 2h, 4h and 6h, SF group SEC ICAM-1 concentration, respectively (44 ± 3.4 pg / ml, 68 ± 3.2 pg / ml and 73 ± 4.5 pg / ml) was significantly lower than the control group ( 59 ± 4.3 pg / ml, 98 ± 6.5 pg / ml and 111 ± 5.7 pg / ml, P lt; 0.05); the SF group SEC's NF-κB expression was significantly weaker than the control group. Conclusion: SF can reduce IRI transplanted liver especially antral endothelial cell damage, and its mechanism is through the inhibition of NF-κB activation in the liver tissue, the effective cut of TNF-α secretion, decreased generation of ICAM-1, thereby reducing the leukocyte adhesion to the SEC, a significant improvement of microcirculation. This study may provide a new way to improve the quality of liver transplantation.