| Objective: Established the urine derived sepsis rabbit model; application the NaHS(the H2S donor) intervention, investigate the effect of hydrogen sulfide on the role ofurine derived sepsis acute renal injury, and the influence of cell apoptosis of renaldamge,NF-κB,Bcl-2,Bax expression, and then reveal H2S effect on urinary derivedsepsis kindey cells apoptosis.Method:1.Take the ligation of the rabbits left ureter, and to the ligation of theproximal ureteral injection the Escherichia coli (ATCC25922), the rabbit sepsismodel was established because of the acute upper urinary tract obstruction andinfection.2.48healthy male rabbits (weight:1.8-2.2Kg/one) were randomly dividedinto4groups,12rats in each group. The control group, sham operation group,Urosepsis group(model group),NaHS treatment group;3.Each animal according to thetime points were divided into the preoperation,12h after the surgery,24h after thesurgery, postoperative36h, and postoperative48h,5time points observed;Observationthe RT, RR, HR;4.Before and after the operation of each time points, collecting theblood, peripheral blood (WBC, neutrophils)、 renal function (Cr, BUN)test;5.Postoperative48hours take the kidney tissue of rabbits of each group by HEstaining, the morphological changes were observed under light microscope,renal cellapoptosis detection by Tunel; Every time points and the every group take the kindeytissue were detected by immunohistochemical method of Bcl-2, Bax proteinexpression;Western-Blot was dected the kindey tissue NF-kappaB protein expression.Result:1.Comparison of the rabbit body temperature, heart rate, respiratory frequency:control group and sham group showed no significant (P>0.05);model group has themore significantly than group control and sham (P＜0.05); the treatment group waslower than model group (P <0.05).2. The blood test results show: The rabbits of each preoperative group (the bloodleukocytes, neutrophils), creatinine (Cr), blood urea nitrogen (Bun), there was nosignificant difference. Postoperative blood WBC, neutrophil; Cr, Bun results: The control group compared with sham group, there was no significant (P>0.05);modelgroup has the more significantly than group control and sham (P <0.05); treatmentgroup and model group compared to a downward trend, with statistical significance (P<0.05).3.Pathomorphological:The micro morphological changes: light microscope (HEstaining):model group showed glomerular atrophy, deformation, and note the amountof inflammatory cell infiltration; treatment groups compared with model groupreduced the change.4. Immunohistochemistry method has to check the expression of renal tissue Bcl-2,Bax protein: control group, sham group showed renal tissue Bcl-2, Bax proteinexpression was weak,Bax protein expression in model group was the strongest, whileBcl-2protein expression in model group was weak; significantly decreased expressionof treatment group than in model group of Bax protein and Bcl-2protein expression.5. TUNEL assay in renal cell apoptosis: control group kidney tissue in sham groupscattered in a small amount of apoptosis of renal tubular epithelial cells. The modelgroup showed apoptosis film and formation of apoptotic bodies, compared with thecontrol group(P<0.05).treatment group than in model group significantly reduce renalcell apoptosis (P<0.05).6. Western-Blot was detected the NF-kappaB protein expression: group control andsham there was no significant difference in the detection of NF-kappaBprotein;control group compared with model group: the detection of NF-kappaBprotein after48hours had significant difference (P<0.05) was statistically significant;compared with group treatment and model:NF-kappaB protein was detected in48hours after the operation has the significant difference (P<0.05) was statisticallysignificant. Conclusion: Hydrogen sulfide can inhibit the NF-κ B and the Bax expression,increase the Bcl-2expression, reduce the kindey cell apoptosis, reduce the kindeydamage. |
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