Effects Of Sodium Acetated Ringer’s Solution On Cytokines And NF-κB Signaling Pathway In Liver Tissue Of Rats With Hemorrhagic Shock

Objective:To investigate the effects of acetate ringer’s solution on the cytokines and nuclear factor-κB(NF-κB)signaling pathways in liver tissue of rats with hemorrhagic shock.Methods:Thirty-two healthy Sprague Dawley(SD)rats were used to establish a hemorrhagic shock model.They were randomly divided into four groups: shock-free resuscitation group(CR group,n=8),saline resuscitation group(NR group,n=8),lactate ringer’s solution resuscitation group(LR group,n=8),and acetated ringer’s solution resuscitation group(AR group,n=8).After 1 h of shock in NR group,LR group and AR group,different resuscitation fluids(saline,lactate ringer’s solution and acetate ringer’s solution)were used for restrictive fluid resuscitation in shock rats.The recovery method was as follows: firstly,the average arterial pressure(MAP)of the shock rats was resuscitated to 60 mmHg by using different resuscitation fluids,and the MAP was further raised to 80 mmHg to correct the shock after 30 minutes.After the completion of the restrictive resuscitation,the rats in each group were observed for 4 hours,and the CR group continued to maintain the shock state of the rats.Only after the shock was induced,the liver tissues were taken for 4 hours(if the rats died,they were obtained immediately).After liver tissue was fixed in 4% paraformaldehyde solution,HE staining and pathological changes were observed under microscope.Real-time quantitative PCR was used to detect the expression levels of TNF-α mRNA,IL-4mRNA and IL-10 mRNA in liver tissue.Western blot analysis was used to detect the expression of p65(Ser536)phosphorylation and p65(Lys310)acetylation in liver tissue.Results:1.Liver histopathology showed that the edema swelling of the hepatocytes in the AR group was significantly reduced compared with the CR group,the NR group and the LR group,the inflammatory cell infiltration was less,the hepatocytes were less bruised,and no obvious hepatocyte necrosis was observed.2.Compared with CR group(10.41±5.45),NR group(10.16±3.71)and LR group(7.87±2.58),the expression level of TNF-α mRNA(5.06±2.59)in liver tissue of AR group was decreased(P<0.05).3.Compared with CR group(0.69±0.31),the expression level of IL-4 mRNA(7.26±3.03)in liver tissue of AR group was significantly increased(P<0.01).Compared with NR group(3.89±2.74)and LR group(4.33±1.96),the expression of IL-4 mRNA in liver tissue of AR group increased(P <0.05).4.Compared with CR group(0.81±0.53)and NR group(4.04±2.40),the expressionlevel of IL-10 mRNA(9.33±3.79)in liver tissue of AR group was significantly increased(P<0.01).Compared with the LR group(4.97±2.46),IL-10 mRNA expression was increased in the liver of the AR group(P <0.05).5.Compared with CR group(0.44±0.13)and NR group(0.33±0.06),the expression level of p65(Ser536)phosphorylation(0.18±0.11)was significantly decreased in AR group(P<0.01).Compared with the LR group(0.27 ± 0.04),the phosphorylation level of p65(Ser536)was decreased in the liver of the AR group(P<0.05).6.Compared with CR group(0.40±0.09)and NR group(0.30±0.05),the expression level of p65(Lys310)acetylation(0.15±0.10)in liver tissue of AR group was significantly decreased(P<0.01).Compared with LR group(0.24±0.05),the expression level of acetylation of p65(Lys310)in liver tissue of AR group was decreased(P<0.05).Conclusion:1.Different crystalloids were used for early restrictive fluid resuscitation in hemorrhagic shock rats.Acetate Ringer’s solution can improve the pathological changes of liver tissue and reduce it further than conventional resuscitation fluid(salt saline and lactate ringer’s solution).The release of pro-inflammatory factor TNF-α from liver tissue and the early expression levels of anti-inflammatory factors IL-4 and IL-10 enhance liver injury in hemorrhagic shock rats.2.Rats with early restrictive resuscitation of hemorrhagic shock with acetate ringer’s solution could inhibit the expression of p65(Ser536)phosphorylation and p65(Lys310)acetylation in liver tissue.Therefore,it is possible to reduce the release of pro-inflammatory factors and increase the level of anti-inflammatory factors by inhibiting the activation of the NF-κB signaling pathway,thereby reducing the liver damage induced by shock in rats.