Inhibition Of Endogenous Hydrogen Sulfide Increase The Inflammation Of Renal Injure In Urine-dirived Sepsis

Aims To establish the urine-derived sepsis model in rabbits, cystathionine gamma lyase(CSE) inhibitor DL-Propargylglycine(PAG) was used to inhibit the generation of endogenous hydrogen sulfide, observing the changes of endogenous H2 S production and CSE activity, expression of nuclear factor kappa B(NF-κB) in renal tissues and the effect of cell factor on endogenous H2 S, clear urine-derived sepsis and expression of renal cytokines, which reveals the possible mechanism of the role of H2 S playing in human urinary septic renal damage in pathological process.Methods 1, 32 healthy male rabbits were selected, randomly divided into 4 groups, 8 rabbits in each group, respectively: normal(control) group; sham operation group(sham); urine-derived sepsis group(sepsis); DL- Propargylglycine(PAG) group.2, The middle left ureteral ligation of rabbits in group sepsis and to the proximal lumen into Escherichia coli(ATCC 25922), the establishment of urine-derived sepsis rabbit model; group PAG after modeling to intraperitoneal injection of DL- Propargylglycine(37.5mg/kg), group sepsis of intraperitoneal injection of saline.3,All rabbits were further divided into five observation points according to the time of preoperative, postoperative 12 h, 24 h, 36 h and 48 h,, rectal temperature(RT),respiratory rate(RR), heart rate(HR)were record on each time point.4, Blood line(WBC, Neutrophils), renal function(Cr, BUN) were tested in peripheral blood of preoperative, postoperative 12 h, 24 h, 36 h after operation and postoperative 48 h respectively.5, Plasma H2 S concentration was measured and CSE activity of renal tissue were detected postoperative 48 h.6, Rabbit kidney specimens were fixed and HE stained postoperative 48 h, structural changes in morphology were observed under an optical microscope. Expression in renal tissue was detected using Western-blot method of NFkappa B and cell factor TGF- beta 1, IL-6.Results 1, Vital signs:There was no significant difference about RT, RR, HR between sham group and control group postoperative 48h(P>0.05); sepsis group at each time point of RT,RR, HR were significantly higher than that in control group and sham group(P<0.05); PAG group RT, RR, HR than in sepsis group were significantly higher(P<0.05).2, The blood test indexes: There was no significant difference about WBC,Neutrophils, Cr, BUN in sham group compared with control group, 48 h after operation(P>0.05); WBC, Neutrophils, Cr, BUN in sepsis group were significantly higher than those in control group and sham group(P<0.05); WBC, Neutrophils, Cr, BUN in PAG group were significantly higher than in sepsis group(P<0.05).3, Detection of the activity of CSE and the content of plasma H2 S in renal tissue: there was no significant difference between sham group and control group of renal tissue CSE activity and H2 S content in plasma after 48h(P>0.05); Compared with control group and sham group,renal tissue CSE activity and H2 S content of sepsis group were decreased(P<0.05);Renal tissue CSE activity and H2 S content in PAG group was significantly decreased than in sepsis group(P <0.05). All rabbits kidney tissue CSE activity was positively correlated with plasma H2 S content relationship, R=0.8505.4, The histological change of pathology: The pathological morphological was HE dyed. The structural form of kidney morphology in control and sham groups of rabbit was in normal form and roughly same. The abscess was visible in kidney tissues in Group sepsis, with a large number of inflammatory cell infiltration, renal pelvis and calyces mucosa falling off. Glomerular and renal capsule come to change and renal tubular was altered. Group PAG was more serious than that in sepsis group.5, Expression of NF-kappa B, IL-6 and TGF- beta 1 protein in renal tissue(Western-blot method): compared with control group,the expression of in renal tissue of NF- kappa B, IL-6 and TGF- beta 1 in sham group postoperative 48 h showed no significant difference(P>0.05); the expression of NF- kappa B, IL-6, TGF- beta 1 sepsis group and PAG group were higher than those in control group(P<0.05); the expression of NF-kappa B, IL-6 in PAG group increased than in sepsis group(P<0.05), TGF- beta 1 expression decreased than sepsis group(P<0.05).Conclusions 1. A urine-derived sepsis rabbits model was successfully established in this study.2. Application of PAG can inhibit the generation of endogenous hydrogen sulfide and increase renal injury induced by urine-derived sepsis, probably via enhancing the expression of NF- κB and IL-6 as well as decreaseing the expression of TGF-β1.