| Aims To establish a Urosepsis rabbit model. Applications sodium bisulfide and PAG intervention,observe the change of ET-1,MAD and SOD, which reveals the mechanism of H2S in the urine-derived sepsis with renal damage.Methods1 To establish the rabbit urospsis model by the intracornoary infusion of autologous cecal contents and ATCC 25922 to the left ureter, and afterward ligation of it. Thirty-five of healthy male rabbits randomly were divided into five groups?N=7.each?.group control?normal control group?;group sham?sham operation group?;group sepsis?urosepsis group?;group Na HS?injected 8.4umol/kg Na HS by ear vein?; group PAG? intraperitoneal injection of PAG?.2 Animals in each group divided into preoperative and postoperative 12 h, postoperative 24 h, postoperative 36 h postoperative 48 h,5 time observation point by point in time. Respectively, at each time point observation anal temperature? RT?, respiratory? RR?, heart rate?HR?;3 blood samples were collected at preoperative, postoperative 12, 24, 36 and 48 hours. peripheral blood cell count?WBC, neutrophil?, renal function? Cr, BUN? were detected;4 The pathological changes of kidney tissue with HE were observed by optical microscope and electron microscopic in 48 h after operation.5 The ET-1and SOD m RNA expression in the kidney tissues were detected by RT-PCR in 48 h after operation;6 The ET-1 protein expression were detected by Western Blot,SD and MAD activities were measured by spectrophotometry in 48 h after operation;7 The acatives of CSE and the concentration of H2S were test in 48 h after operationResults1 The vital signs of ribbit: Results on RT?RR and HR between group sham and group control shows no statistically significant differences?P>0.05?;group sepsis is significantly higher than that in group control and sham?P<0.05?; Compared with group sepsis, the RT?RR and HR were increased obviously in group PAG?P<0.05?; At any time points, the RT?RR and HR of group Na HS is lower than that in group sepsis and PAG?P< 0.05?, while higher than in group control and sham?P< 0.05?.2 Peripheral blood cell count:Compared with control group, N were increased in Sham group in 12 h and 24 h after operation?P<0.05?; The WBC and N of Sepsis group were higher than Control and Sham group?P<0.05?;.Compared with group sepsis, the WBC and N were increased obviously in group PAG?P<0.05?; At any time points, the WBC and N of group Na HS is lower than that in group sepsis and PAG?P< 0.05?, while higher than in group control and sham?P< 0.05?.3 Kidney function: Results on Cr and Bun between group control and group sham show not statistically significant differences?P>0.05?;group sepsis is significantly higher than that in group control and sham?P<0.05?, and group PAG is higher group sepsis?P<0.05?, Compared with group Sepsis and PAG, group Na HS is decreased, and group Na HS is significantly higher than that in group Control and Sham?P<0.05??4 Pathological change: pelvis, calyceal mucosal necrosis, shedding massive neutrophil infiltration, the glomerular renal capsule deformation, tubular lumen, renal interstitial inflammatory cell infiltration; glomerular transmission electron microscopy group sepsis fusion of podocytes, disappear, mesangial cells and endothelial cell proliferation, mitochondrial cristae disorder,vacuolation. Group PAG than in the group sepsis of the above changes have augmented group while Na HS than in the group sepsis of the above changes have abated.5 ET-1 and SOD expression: ET-1 and SOD expression in the kidney tissues in 48 h after operation show no statistically significant differences in control and sham group?P>0.05?;RT-PCR results showed the ET-1 m RNA expression in Sepsis group increased compared with Control and Sham group?P<0.05?;Compared with Sepsis group, the ET-1 m RNA expression in PAG group further increased?P<0.05?.The m RNA expression of ET-1 decreased in Na HS group compared with Sepsis and PAG group?P<0.05?;the SOD m RNA expression in Sepsis group decreased compared with Control and Sham group?P<0.05?;Compared with Sepsis group, the SOD m RNA expression in PAG group further decreased?P<0.05?.The m RNA expression of SOD increased in Na HS group compared with Sepsis and PAG group?P<0.05?.6 MDA, ET-1 and SOD activities: MDA, ET-1 and SOD expression in the kidney tissues in 48 h after operation show no statistically significant differencesin control and sham group?P>0.05?;The ET-1 and MDA activities in Sepsis group increased compared with Control group?P<0.05?;Compared with Sepsis group,the ET-1 and MDA activities in PAG group further increased?P<0.05?.The m RNA expression of ET-1 and MDA activities decreased in Na HS group compared with Sepsis and PAG group?P<0.05?;In the five group, the SOD activities of PAG group was the lowest?All P<0.05?;Compared with control and sham group, the SOD activities in Sepsis group decreased?P<0.05?.The SOD activities increased in Na HS group compared with Sepsis group?P<0.05?.7 Plasma hydrogen sulfide concentration and CSE in renal tissue : hydrogen sulfide concentration and CSE in 48 h after operation show no statistically significant differencesin control and sham group?P>0.05?;CSE activity in Group Na HS and group Sepsis were decreased compared to control and sham group?P<0.05?. Compared with Sepsis and NHS group,the CSE activity in PAG group further decreased?P<0.05?.Compared to Control and Sham group, plasma hydrogen sulfide concentration were obviously decreased in group Sepsis,and further decreased in group PAG?P < 0.05?.Conclusions1 The the rabbit urospsis model is established successfully.2 PAG can inhibit the production of endogenous H2S and further aggravated kidney injury. The exogenous H2S that were produced by applying sodium bisulfide can alleviate kidney injury induced by urine-derived sepsis.3 H2S plays a protective role in the urine-derived sepsis-induced AKI. The mechanisms of actions may be attributed to its modulating effects that inhibits the ET-1 expression, downregulated MDA activity and enhanced the SOD activity. |
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