| Objective: Through the establishment of rat renal ischemia-reperfusion injury models to study the effect of renal ischemia-reperfusion injury in rats after renal ischemia reperfusion injury, and explore the possible protective mechanism of mediators on renal ischemia reperfusion injury in rats.Methods: We build three models of renal ischemia reperfusion injury in rats were divided in renal ischemia reperfusion injury group without irrigation(group A), renal ischemia reperfusion injury group with irrigation(group B) and renal ischemic reperfusion injury group without nephrectomy of right kidney(group C) and sham operation group(group D). Group A was treated by removing the right kidney, left kidney treated with ischemia and 60 min after reperfusion for 24 h, by group B the right kidney was removed and the left kidney was lavaged at first, then ischemia for 60 min and 24 h reperfusion, by group C only left kidney treated with ischemia for 60 min then reperfusion for 24 h, group D treated with open, closing the abdomen. The inferior vena cava blood and kidney samples were detected by sacrifice: 1. The serum creatinine and blood urea nitrogen levels were detected by automatic biochemical analyzer; 2. Enzyme linked immunosorbent adsorption assay(ELISA) method to detect the monocyte chemotaxis protein-1(MCP-1), tumor necrosis factor alpha(TNF-α), interleukin–4(IL-4) and interleukin-6(IL-6) levels; 3. Routine embedding, slice and PAS staining, the pathological changes of renal tissue and inflammatory infiltration were observed in each group; 4. The immunohistochemical method to detect the monocyte chemotaxis protein-1(MCP-1), tumor necrosis factor alpha(TNF-α) and interleukin-4(IL-4) and IL-6(IL-6) expression; 5. Fluorescence quantitative PCR method to detect the the monocyte chemotaxis protein-1(MCP-1), tumor necrosis factor alpha(TNF-α) and interleukin-4(IL-4) and IL-6(IL-6) m RNA expression.Results: 1. The automatic biochemical analyzer to detect the levels of serum creatinine and urea levels are significantly increased in group A and group B, and group A was significantly higher than that of group C and group D(P<0.01), B group were significantly higher than those in group C and group D(P<0.01), and group A and group B no significant difference, group C and group D had no significant difference; 2. ELISA method to detect MCP-1, TNF-α, IL-4, IL-6, group D was little, group A, B and C were increased significantly, group C than in group A and group B decreased significantly(P<0.01), A and B two groups had no significant difference(P>0.05); 3. Renal tissue PAS staining pathology examination showed: serious damage and disorder of group A renal tubules, renal interstitial edema and inflammatory cell infiltration, renal tubular gap was increased, lumen expansion, renal tubular brush border loss, tubular epithelial cells were flat, visible off, bare membrane; group B of renal tubular structure is more clear, renal interstitial edema and inflammatory cell infiltration compared with the group A, the renal tubule brush border intact tubular epithelial cell swelling, degeneration, occasionally shedding or canal; serious damage and disorder of group C left renal tubular structure, renal interstitial edema and inflammatory cell infiltration, renal tubular gap was increased, lumen expansion, renal tubular brush border loss, tubular epithelial cells were flat, visible off, bare membrane; group C of right renal tubules, basement membrane integrity, no interstitial edema, a very small amount of inflammatory cell infiltration in renal tubule; group D The structure was clear, the basement membrane was complete, no interstitial edema and inflammatory cell infiltration were found; 4. Immunohistochemistry showed that MCP-1, TNF-α, IL-4, IL-6 in group C, the right kidney and group D was rarely expressed, in the group A, B and group C, the left renal expression were significantly increased. The expression was mainly in cortex and medulla and outer medulla outer border, group B was lower than that in group A and group C left kidney decreased significance(P<0.01), but still higher than that of group C right kidney and group D(P<0.01), group C: right kidney is higher than that of group D(P<0.01) in group A and group C left kidney had no significant difference(P>0.05); 5. Fluorescence quantitative PCR showed MCP-1, TNF-α, IL-4, IL-6 in C group, the right kidney and group D was rarely expressed, in the group A, B and group C, the left renal expression were significantly increased, group B was lower than that in group A and group C left kidney decreased significantly(P<0.01), but still higher than that of group C right kidney and group D(P<0.01). In group A and group C of the left renal no obvious difference(P>0.05), group C right kidney and group D had no significant difference(P>0.05).Conclusion:Renal ischemia in rats can be reduced by reducing the MCP-1, TNF-α, IL-4, IL-6 and other inflammatory mediators to produce a protective effect on renal ischemia reperfusion injury in rats. |
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