| Objective: In order to explore preliminarily the effect and the related possible mechanism of AMPK signal pathway changes for the rats’ liver ischemia-reperfusion injury,AMPK agonist AICAR has been used for preconditioning the rats’ liver ischemia-reperfusion injury model.Methods: With random and equality,fifty-four SD rats,male,healthy and SPF-rated,were divided into three groups respectively.The experimental group:5-aminoimidazole-4-carboxamide nucleotide(AICAR)+ ischemia for 30 mins.And then AICAR was administered intraperitoneally with the dose of 500 mg/kg an hour ago before surgery.The control group:ischemia-reperfusion+ischemia for 30 mins.And then saline(500 mg/kg)was administered intraperitoneally one hour ago before surgery.Sham-operated group: the preoperative treatment is the same as the control group.Samples were collected after 12,24 and 72 hours respectively after the hepatic ischemia-reperfusion,and the level of ALT,AST,TBil,TNFα and IL-6 were tested respectively.the content of ATP in liver tissues were measured,routine pathological staining(HE staining)was performed to observe the histological damage of the liver,and real-time fluorescence quantitative PCR was performed to detect the expressive levels of adenylate-activated protein kinase(AMPK),mammalian target of rapamycin(mTOR),glucose protein(mTOR),glucose transporter protein type 4(GLUT4),and multidrug resistance protein 2(MRP2)mRNA.Western Blot was performed to detect the protein levels of phosphorylated AMPK,phosphorylated mTOR,phosphorylated GLUT4,and MRP2 in liver tissues.Results: 1.The results of conventional pathological staining(HE staining)of the liver showed that the sham operation group had no obvious liver tissue and structural damage at various points in time;the liver damage in the control group and the experimental group was heavier than the sham-operated group at various points in time group,and the liver damage in experimental group and in the control group was worse at 24h than at 12h after surgery,and the liver damage at 72h was less than at 24h.The degree of liver damage in the experimental group was less than that of the control group at all time points.2.Serological results showed: the serum of ALT,AST,TBil,IL-6,and TNF-α of the control group were higher than those of the experimental group and the sham operation group,while that of the experimental group was higher than that of the sham operation group after 12,24,and 72 hours after operation;From 12h to 24h after operation,the changes of each index increased,while from 24h to 72h,the changes of each index decreased.3.The results of the liver tissue enzyme-linked immunosorbent assay showed: after operation,the ATP content of liver tissue in the experimental group was higher than that of the control group and the sham operation group at 12,24,and 72 hours,while that of the control group was lower than that of the sham operation group;from 12h to 72h,the content of ATP of liver tissue in the control group showed a rising trend;and the content of ATP of liver tissue in the experimental group increased from 12h to 24h after surgery,but there was no obvious change in ATP content at 72h and 24h after surgery.4.PCR and Western Blot results showed: After operation,the relative expression levels of AMPK,GLUT4,mRNA of MRP2,phosphorylated AMPK,phosphorylated GLUT4,and protein of MRP2 in experimental group were higher than those in control group and sham operation group;The relative expression levels of mRNA of mTOR and its phosphorylated protein in the experimental group were lower than those in the control group and the sham operation group.Conclusion: AICAR pretreatment can activate the AMPK signaling pathway,improve the metabolic pathways of energy,and reduce the hepatic inflammatory response,so that the extent of HIRI can be reduced. |
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