| Aims To establish a model of urine-derived sepsis in rabbits with acute upper urinary tract obstruction and infection and to study the changes of iNOS and HO-1 in renal tissue during sepsis and the effects of Na HS and PAG on urine-derived sepsis rabbit model to further clarify the H2 S on urine-derived sepsis in rabbits kidney tissue iNOS,HO-1 expression,to explore the H2 S in urine-derived sepsis acute renal injury in rabbits and its protective effect Possible mechanism.Methods 1?40 healthy male rabbits were randomly divided into 5 groups(n = 8),which were set as Control group,sham group,Sepsis group,Sodium Hydrosulfide(Na HS)group and DL-propargylglycine(PAG)group.2?In the urine-derived sepsis group,hypertensive renal pelvis was used to simulate upper urinary tract obstruction and infection.The rabbit was ligated to the distal end of the left ureter and the ureter was injected into the upper ureter and injected into E.coli(ATCC25922);PAG group The rabbits were injected intraperitoneally with DL-propargylglycine solution(37.5mg/kg)based on the above model;the sodium hydrosulfide group was injected with Na HS solution(Na HS 8.4?mol/kg)via the ear vein of rabbits on the basis of modeling.In the sham group,only the left ureter was dissociated and the normal control group was not treated.3?The rectal temperature(RT),respiratory rate(RR)and heart rate(HR)were measured before and 24 hours after operation,48 hours after operation,and 72 hours after operation in the five groups of rabbits.4?The number of white blood cells(WBC),neutrophil(N)in four groups of rabbits at 4 time points(preoperative,postoperative 24 h,postoperative 48 h,postoperative 72h),C-Reaction protein(CRP),creatinine(Cr)and blood urea nitrogen(BUN).5 ? The left kidney of rabbits were stained with HE at 72 h after operation,and the histopathology and morphological changes of renal tissues were observed by light microscope.6?The expression of iNOS and HO-1 protein in kidney of each group were detected by Western blot at 72 h after operation.7?The relative expression of iNOS m RNA and HO-1m RNA in renal tissue of rabbits in each group was determined by reverse transcription-polymerase chain reaction(RT-PCR)method at 72 h after operation.Results: 1?vital signs: There was no significant difference in the preoperative RT,RR and HR between the two groups(P> 0.05).There was a slight increase of HRT,RR,HR in Sham group compared with control group at the same time point(P <0.05).The RT,RR and HR tended to be normal in Sham group at 48-72 h postoperatively,with no significant difference compared with Control group at the same time point(P> 0.05).The RT,RR and HR of Sepsis group were gradually(P <0.05)compared with control group and Sham group at the same time.The RT,RR,HR of PAG group were significantly higher than those of Sepsis group at different time points(P <0.05).The levels of RT,RR and HR in Na HS group were significantly lower than those in Sepsis group(P <0.05).2 ? Peripheral blood cell count and CRP results: There was no significant difference between the preoperative WBC,N and CRP levels in the five groups of rabbits(P> 0.05).The levels of WBC,N and CRP in Sham group increased at 24 h after operation,while the WBC,N and CRP tended to be normal at other time points.There was no significant difference between Sham group and control group at the same time point(P> 0.05).The WBC,N and CRP in Sepsis group increased gradually at 24 h,48h and 72 h after operation,which were significantly higher than those in Control and Sham group at the same time points(P <0.05).WBC,N and CRP in PAG group were significantly higher than those in Sepsis group at 24 h,48 h,72 h after operation(P <0.05).Compared with the Sepsis group,the WBC,N and CRP in the Na HS group decreased significantly at all time points(P <0.05)at 24 h,48 h and 72 h after operation.3?Renal function: There was no significant difference between the preoperative levels of Cr and BUN in the five groups of rabbits(P> 0.05).The levels of Cr and BUN in Sham group at 24,48 and 72 h after operation were not different from those in control group at the same time point(P> 0.05).Compared with Control group and Sham group,Cr and BUN in peripheral blood of Sepsis group increased continuously at 24 h,48h and 72h(P <0.05).The levels of Cr and BUN in peripheral blood of PAG group increased significantly at 24 h,48 h,72 h after operation compared with Sepsis group(P <0.05).The levels of Cr and BUN in peripheral blood of Na HS group decreased at 24 h,48 h and 72 h after operation compared with Sepsis group(P <0.05).4?The histopathological changes of renal tissue were observed under a light microscope(HE staining): The morphology of the glomeruli and renal tubules in the Control group and the Sham group had no abnormalities.There was no difference in the morphological structure between the two groups.In the Sepsis group,pathological changes of renal cell morphology,glomerular deformation,edema,hyperemia,and cyst enlargement were evident in the high power microscope.Renal tubular epithelial cells showed edema and necrosis,renal interstitial cell edema,and neutrophil infiltration.Compared with Sepsis group,the histopathology and morphology of renal tissue in PAG group were significantly aggravated.The pathological structure and cell morphology of renal tissue in Na HS group were significantly less than those in the group of Sepsis.5?Expression of iNOS protein and HO-1 protein in renal tissue 72 h after operation: No significant difference was found in the expression of iNOS protein and HO-1 protein in control group and Sham group at 72 h after operation(P> 0.05);The expression of iNOS protein and HO-1 protein in Sepsis group was significantly higher than that in Control and Sham group(P<0.05)at 72 hours after operation.Compared with Sepsis group,the expression of iNOS protein increased in PAG group and decreased in Na HS group at 72 h after operation(P <0.05).Compared with Sepsis group,HO-1 protein expression increased in Na HS group and decreased in PAG group at 72 h after operation(P <0.05).6?72h after operation iNOS m RNA and HO-1 m RNA expression in renal tissue:The expression of iNOSm RNA and HO-1m RNA in control group and Sham group at 72 h after operation were not significantly different(P> 0.05).The expression of iNOS m RNA and HO-1 m RNA in Sepsis group was significantly higher than that in Control group and Sham group at 72 hours after operation(P<0.05).Compared with Sepsis group,the expression of iNOS m RNA increased in PAG group and decreased in Na HS group at 72 h after operation(P <0.05).Compared with Sepsis group,the expression of HO-1m RNA increased in Na HS group and decreased in PAG group at 72 h after operation(P <0.05).Conclusions: In urine-derived sepsis,the expression of iNOS and HO-1 were increased in renal tissue.H2 S could up-regulate the expression of HO-1 by inhibiting the expression of iNOS,which may play a protective role in urine-derived sepsis induced acute kidney injury. |
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