| Objective:In this study,the obstructive nephropathy rat model was established by unilateral ureteral obstruction?UUO?in which the Rennin-angiotensin-aldosterone system?RAS?and the Mineralocorticoid receptor were activated,leading to the inflammatory injury and interstitialfibrosis of the kidney.To observe the expression of cell proliferation the index of PCNA and analysis the causes which may be related to inflammatory mediators and profibrogenic cytokines.By means of observing the expression of the Serum and Glucocorticoid-induced Protein Kinase 1?SGK-1?,and detecting the variation of the index of correlationas Monocyte Chemotactic Protein1?MCP-1?,F4/80,Transforming Growth Factor-?1?TGF-?1?to study the role of eplerenone in resistance to inflammatory injury and interstitial fibrosis and the mechanism of the Huayu Jiedu Decoction in resistance to the interstitialfibrosis of the kidney.Methods:Forty-eight healthy male Wistar rats weighting about 200±20 g were used in this study.The experimental rats ate and drank freely and the temperature was controlled in?22±2??.After one week adaptive feeding,rats were randomly divided into SHAM group,UUO group,EPL group,and ZY group,for 12 rats in each group.Then,all rats were copied with the unilateral ureteral obstruction nephropathy model except for the SHAM group:after anesthesia with 10%chloral hydrate,the rats were sliced through the left middle abdominal skin,separated the left ureter,ligated respectively on ureteral 1/3 and middle 1/3 ligation with silk,cut off the ureter.Then sutured in skin by layer to layer.The rats in SHAM group were only separated the left ureter but not ligated and cut off the ureter.After operation,the rats were treated with 100 mg·kg-1·d-11 Eplerenone in fodder in EPL group and14g·kg-1·d-11 in ZY group;others in SHAM group and UUO group administered with the same volume of saline once a day.Continuous intervention was for 10 days.Beheaded after weigh the rats 10 days later,retain blood specimens for detection.Cut down the obstruction kidney,weigh the tissue after remove integument,part of the kidneys were fixed by 4%paraformaldehyde and part of the kidney kept at-70?for extracting protein.The expression of SGK-1,MCP-1,F4/80,IL-1,TGF-?1,TNF-?,PCNA were detected by the way of immunohistochemistry and Western blot.And the slices were subjected to routine HE and Masson staining.Results:1 The change of Pathomorphology in renal tissueHE staining showed that tubular epithelial cells arranged regularly and it had few inflammation cells in SHAM group.There were a large number of inflammation cells infiltration,and renal tubular epithelial cells were denatured,shed and turgid in UUO group.The inflammation cell infiltration was significantly reduced and renal tubular epithelial cells were turgid in EPL and ZY groups.Masson staining showed that there was a little collagen deposited in renal interstitium in SHAM group,the glomerular and tubular basement membrane were clear to be seen.In UUO group,the tubular interstitial collagen was obviously increased and the renal structure was disordered.There were some degree of collagen deposition in EPL and ZY groups,but it was more observably reduced than UUO group.2 The expression of PCNA,F4/80,MCP-1,IL-1,TNF-?was examined by immunohistochemistryIn the UUO group,the expression of PCNA and F4/80 was significant enhanced compared with the SHAM group?P<0.05?;in EPL and ZY groups,the expression was significant decreased?P<0.05?.In SHAM group,the expression were weak of MCP-1?IL-1?TNF-?mainly in distal renal tubular epithelial cells.In UUO group,the expression was enhanced mainly in renal tubular epithelial cells and interstitium cells.In ELP and ZY groups,the expression were positive but significantly decreased in range and strength.3 SGK-1,MCP-1,F4/80,IL-1,TGF-?1,TNF-?,PCNA were detected by Western blotIn the UUO group,the expression of SGK-1,MCP-1,F4/80,IL-1,TGF-?1,TNF-?,PCNA was significant enhanced compared with the SHAM group?P<0.05?;in EPL and ZY groups,the expression was significant decreased?P<0.05?.Conclusions:1 Obstructive nephropathy can induce Mineralocorticoid receptor activation,stimulate renal inflammatory response and cell proliferation,involved in the formation of renal interstitial fibrosis.2 Eplerenone can reduce the damage of obstructive nephropathy by blocking the pro-inflammatory effect of aldosterone.3 By reducing the expression of SGK-1,MCP-1,F4/80,IL-1,TNF-?and TGF-?1,the Huayu Jiedu Decoction can retard the process of renal interstitial fibrosis as well as inhibit renal inflammatory reaction and interstitial cell proliferation. |
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