The Protection And Mechanism Of Hydrogen Sulfide Urine-derived Sepsis With Acute Kidney Injury

Objective: To establish a Urosepsis rabbit model. To observation of the endogenousH₂S generation of the kidney and its H₂S cystathionine-γ-lyase （CSE） changes.Applications exogenous hydrogen sulfide intervention，explore the role of exogenoushydrogen sulfide in the urine-derived sepsis with acute kidney injury，to understandthe impact of H₂S on urine-derived sepsis, renal tissue expression of NF-κB, whichreveals the mechanism of H₂S in the urine-derived sepsis with renal damage.Method：1.To establish the rabbit urospsis model by the intracornoary infusion ofautologous cecal contents and ATCC25922to the left ureter, and afterward ligation ofit.2.Thirty of healthy male rabbits randomly were divided into fivegroups（N=6.each）.group control（normal control group）;group sham（sham operationgroup）;group sepsis（urosepsis group）;group NaHS2.8umol/kg（NaHS group,2.8umol/kg）; group NaHS8.4umol/kg（NaHS group,8.4umol/kg）.3animals in eachgroup divided into preoperative and postoperative24h, postoperative48h,postoperative72h4time observation point by point in time. Respectively, at eachtime point observation anal temperature （RT）, respiratory （RR）, heart rate （HR）;4blood collected by preoperative, postoperative24,48, and72hours. peripheral bloodcell count （WBC, neutrophil, PLT）, renal function（Cr, BUN）, serum C-reactiveprotein were analyed;5.The serum levels of TNF-αI and L-10were record andanalyzed by ELISA every points. blood culture and serum Limulus test weredetected after operation72h.6. The pathological changes of kidney tissue with HEwere observed by optical microscope and electron microscopic.Immunohistochemistry（IHC） method was used to assess the protein expression ofTNF-α, IL-10and NF-κB in rabbit renal tissue in every group. Renal cell apoptosiswas test by Tunel.7.The acatives of CSE and the concentration of H₂S were test in72h after operation. Result:1. The general status of ribbit: Results on T and RR between group sham andgroup control shows no statistically significant differences（P＞0.05）；group sepsisis significantly higher than that in group control and sham（P＜0.05）; groupNaHS2.8umol/kg and group NaHS8.4umol/kg is lower than that in group sepsis（P＜0.05）.2. Bloodtest index results: Results on blood of WBC and Neutrophil, PLT, CRP,Cr and Bun between group control and group sham show not statistically significantdifferences（P＞0.05）；group sepsis is significantly higher than that in group controland sham（P＜0.05）.Compared with group group NaHS2.8umol/kg and groupNaHS8.4umol/kg is decreased, and group NaHS8.4umol/kg is significantly lower thanthat in group NaHS2.8umol/kg（P＜0.05）。Serum of TNF-α，IL-10resuls: groupcontrol and group sham are not statistically significant differences（P＞0.05）;Compared with that of group control, group sepsis is significantly higher（P＜0.05）；Compared with that of group sepsis， the serum of TNF-α in groupNaHS2.8umol/kg and group NaHS8.4umol/kg is significantly decreased（P＜0.05），the serum of IL-10is opposite with TNF-α。Serum tal was test by Tunel in72h.Group control and group sham are not statistically significant differences（P＞0.05）;Compared with that of group control, group sepsis is significantly changer（P＜0.05）； The Limulus test than the blood culture-positive rate （P=0.009）.3.Pathomorphological: left kidney naked eye view; HE staining under lightmicroscopy examination; kidney tissue electric mirror transmission:Group control and group sham in renal tissue morphology. Group sepsis Eyeview kidney congestion and edema, multiple abscesses, renal pelvis dilatationempyema, mucosal congestion, hemorrhage, edema, medullary visible yellow stripe-like lesions extend to the cortex. Perirenal tissue congestion and edema, renal hilarlymph nodes was swollen. Light microscopy: pelvis, calyceal mucosal necrosis,shedding massive neutrophil infiltration, the glomerular renal capsule deformation,tubular lumen, renal interstitial inflammatory cell infiltration; glomerulartransmission electron microscopy group sepsis fusion of podocytes, disappear, mesangial cells and endothelial cell proliferation, mitochondrial cristae disorder,vacuolation. group NaHS2.8umol/kg, group NaHS8.4umol/kgp than in the groupsepsis of the above changes have abated.4.TNF-α, IL-10and NF-κB protein IHC results: TNF-α、IL-10and NF-κBstaining is noted in group control and sham；The TNF-α and NF-κB proteinexpression is enhaneed in group sepsis，while group NaHS2.8umol/kg and groupNaHS8.4umol/kg have lower expression than that in group sepsis；The IL-10proteinexpression is opposite with TNF-α.5. Renal cell apoptosis: The apoptosis of renal cel were litile in group control andsham. Compare with than in group control, the group sepsis is significantly change（rP＜0.05）. Compare with than in group sepsis, the group NaHS2.8umol/kg and groupNaHS8.4umol/kg is significantly changer（P＜0.05）, and group NaHS8.4umol/kghave lower expression than that ingroup NaHS2.8umol/kg（P=0.02）.6. Detection of72hours of plasma hydrogen sulfide concentration and CSE inrenal tissue after6: activity of sepsis in renal tissue H₂S content and plasma CSEcompared with control group decreased significantly （P<0.05）. NaHS2.8umol\/kggroup, NaHS8.4umol\/kg group, plasma H₂S levels were significantly elevatedcompared to sepsis group （P<0.05）. NaHS2.8umol\/kg and NaHS8.4umol\/kg group,plasma H₂S content compared with control group, P>0.05. NaHS2.8umol\/kg in renaltissue of group, NaHS8.4umol\/kg activity in CSE group compared with sepsis group,P>0.05. Sepsis in renal tissue of group NaHS2.8umol\/kg and NaHS8.4umol\/kgactivity in CSE group compared with control group, P <0.05. Group Control, groupsham, group sepsis H₂S content in plasma and renal tissue CSE activity waspositively （R=0.979, P <0.01）, and CSE activity of endogenous H₂S positivecorrelation.Conclusion:1. We infuse the left ureter with autologous cecal contents and ATCC25922,then ligate it above the injection points for72hours,so the the rabbit urospsis modelcan established successfully.2. The experimental result found that H₂S can effectively control sepsis and relieve reaction of tissues and organ. 3.The possible mechanism of H2S effectively control sepsis is it can effectivelyprevent and control sepsis events by down-regulate TNF-α, and up-regulate IL-10through NF-KB pathway.