The effect of necrostatin-1 on high mobility group protein-B1 in liver of rats with hemorrhagic shock

Abstract

Objective To investigate the effect and mechanism of necrostatin-1(Hec-1) on the level of HMGB-1 protein in liver of rats with hemorrhagic-traumatic shock. Methods A number of 96 male SD rats were divided into sham-operated group, dimethyl sulfoxide(DMSO) group and Nec-1 group (n=32 in each) by randomized number method. Rat model of hemorrhagic-traumatic shock was made by fracture of femoral bone and tibia bone and exsanguination from femoral vein until 30 mmHg and maintained at 30-40 mmHg for 90 min, then the shed blood was transfused back with Ringer's solution. The rats in sham-operated group were only under anesthesia for separating and ligating blood vessels, without exsanguination to induce hemorrhagic shock and without replenishment with blood. Rats in Nec-1 group were given 1 mg/kg Nec-1 through femoral vein 5 min before replenishment with blood and Ringer's solution, while the rats in DMSO group were given equal volume of DMSO solution instead. Eight rats in each group were sacrificed separately at 2 h, 8 h, 16 h and 24 h after replenishment. The serum and liver tissues of rats in each group were collected to detect serum alanine aminotransferase(ALT) and aspartate aminotransferase(AST), and to observe the pathological changes in liver with hematoxylin-eosin(HE) staining. The level of HMGB-1 in serum was detected by using ELISA. The cytoplasm protein and total protein expressions of HMGB-1 were assessed by using western blot analysis. Results Compared with DMSO group, levels of serum ALT at 8 h(P<0.05), 16 h(P<0.01) and 24 h(P<0.01) in Nec-1 group were significantly lower. Level of serum AST in Nec-1 group were lower compared with DMSO group at 8 h(P<0.01), 16 h(P<0.01) and 24 h(P<0.01). Compared with DMSO group, levels of serum HMGB-1 at 8 h(P<0.05), 16 h(P<0.01) and 24 h(P<0.01) in Nec-1 group were significantly lower. Under light microscopy and transmission electron microscope, hepatic lobule destroyed, the blood extravasated, the immunocyte infiltrated and cellular organelle destroyed were found. Compared with DMSO group, the level of HMGB-1 protein in cytoplasm protein in Nec-1 group were significantly decreased at 8 h(P<0.01), 16 h(P<0.01) and 24 h(P<0.01). The level of HMGB-1 protein in total protein in Nec-1 group were significantly decreased 8 h(P<0.05) and 24 h(P<0.05). Conclusions Nec-1 can remarkably protect the liver of rats with hemorrhagic-traumatic shock, decrease the level of HMGB-1, and protect the hepatocyte effectively. Key words: High mobility group protein B1; Trauma; Shock, Hemorrhagic; Necrostatin-1; Necroptosis