| Objective: To study the effect of PKC beta inhibitor on the expression M1 and M2 macrophage surface markers and their mediator in renal tissue through the establishment of renal ischemia-reperfusion injury models in rat.And provide theoretical basis for the further study of mechanisms of PKC beta inhibitors relieve renal ischemic-reperfusion injury.Methods: Renal ischemia-reperfusion injury model in rats were made and divided into four groups: group A renal ischemia reperfusion injury without pre-irrigation,renal group B ischemia reperfusion injury group with pre-irrigation,group C PKC beta inhibitor treatment and group D sham operation.On the first day,the rats of group A,B and D gavage normal saline 1ml everyone,but the rats of group C using PKC beta inhibitor instead.The second day,in group A,B,C rats were removed the right kidneys,and the left kidneys reperfusion 24 h after ischemia 60 min,before this the left kidneys of group B rats was lavaged with normal saline.Rats in group D only treated with opening,closing the abdomen.24 h later we collect the kidney specimens for testing: 1.The immunohistochemical method to detect the inducible nitric oxide synthase(iNOS),interleukin-12(IL-4)expression analysis of M1 macrophages infiltration in the kidney;2.The expression of macrophage markers(CD197/CD163)were detected with immunofluorescence method to analyse the infiltration of M1 /M2 macrophages in the kidney;3.Fluorescence quantitative PCR method to detect the dendritic cells related type C plant haemaglutinin 1(Dectin-1),Arginase-1(Arg-1)mRNA expression analysis of M2 macrophages infiltration in the kidney.Results: Immunohistochemistry showed that iNOS is mainly expressed in the proximaltubule epithelial cell cytoplasm and capsule,but IL-12 express in the renal medullary renal tubular interstitial,partly in the glomerular mesangial cells and renal tubular epithelial cells in the cytoplasm.The expression of iNOS and IL-12 in the group D is very few,the group A and group B significantly more than group C(P<0.01);similarity,the group A has obviously greater than group B(P<0.01);Contrast with group C,the expression of iNOS levels of group D is decreased(P<0.01),but the expression of Il-12 levels has no obvious change(P>0.05).2.Immunofluorescence showed that CD197 and CD163 are mainly expressed in the renal tubular interstitium.The expression of CD197 in group D is very low,the expression in group A is significantly more than in group B and group C(P<0.01);and there was no significant difference between group B and C(P>0.05).CD197 expression in group D were significantly lower than those in group A,B and C(P<0.05),The expression of CD163 in group C were significantly higher than those in group A,B and D(P<0.01).There was no significant difference between group A,B and D(P>0.05).3.Fluorescence quantitative PCR showed that the expression of Dectin-1 and Arg-1's mRNA in the group D is very few,but in the group C the left renal expression were significantly increased.Dectin-1 and Arg-1's mRNA in group C were significantly higher than those in group A,B and D(P<0.05),and there was no significant difference between group A,B and D(P>0.05).Conclusion: By pre-irrigation before renal ischemia in rats the number of M1 macrophages infiltration can be reduced.PKC beta inhibitor can inhibit the infiltration of M1 macrophages and enhance the expression of anti-inflammatory M2 macrophages to promote tissue repairing and to reduce inflammation after kidney ischemia-reperfusion injury.But the specific mechanism is not clear,it will be confirmed by further research. |
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