Effects Of Shenluotong Decoction On Cell Proliferation In Rats With Unilateral Ureteral Obstruction

Objective: To observe the expression of marker for renal cell proliferation-proliferating cell nuclear antigen?PCNA?,and detect the contents of serum and glucocorticoid-induced protein kinase-1?SGK-1?and transforming growth factor-?₁?TGF-?₁?which were related to cell proliferation,thus investigating the mechanism and the role of Shenluotong Decoction on inhibiting renal cell proliferation and providing the evidence for the clinical application.Methods: 48 healthy clean female Wistar rats weight about 200±20g were used in this study.All the experimental rats ate and drunk freely,and the temperature of laboratory was controled in?22±2??.After one week to adapt,all the rats were randomly divided into four groups: Sham group?n=12?,UUO group?n=12?,EPL group?n=12?,SLT group?n=12?.After anesthesia with 10% chloral hydrate,the rats in UUO group,EPL group and SLT group were sliced through the left abdominal skin,separated the left ureter,ligated respectively on ureteral 1/3 and middle 1/3 with silk,cut off the ureter,then sutured skin.While,the left ureter of rats in Sham group were only separated without ligating or cutting off.After operation,the rats in EPL group and SLT group were respectively treated with eplerenone 100mg·kg-1·d-1 and Shenluotong Decoction 26g·kg-1·d-1,at the same time,the rats in the other groups recevied the same volume of physical saline.Continuous intervention was for 10 days.Cut down the left kidney and weighed it.Part of the kidneys were fixed by 4% paraformaldehyde and the others were kept in-70 ?for extracting protein.The expression of PCNA,SGK-1 and TGF-?₁ were detected by immunohistochemistry and Western blot.Results:1 The change of pathomorphology in renal tissue with HE,Masson and Sirius red stainingHE staining showed that kidney of rats in sham group was normal,there was no edema in renal interstitial with few inflammation cells infiltrated,the tubular epithelial cells arranged regularly without swelling.In UUO group,there were large numbers of inflammatory cells infiltrated in renal interstitium,the renal tubular expanded dramatically and epithelial cells shed.The expansion of renal tubular and the exfoliation of epithelial cells in EPL and SLT groups were the same as UUO group,but the infiltration of inflammation cells was significantly lighter than UUO group.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 SLT groups,but it was more observably reduced than UUO group.Sirius red staining showed that a little collagen deposited in tubular basement membrane and renal interstitiumt of Sham group.While in UUO group,the renal structure was disordered and the tubular interstitial collagen was significantly increased.In EPL and SLT groups,there were still some degree of collagen deposition,but it was notably reduced than UUO group.2 The expression of PCNA?SGK-1?TGF-?₁ by immunohistochemistryThe result of PCNA showed that there was a small amount of positive cells expressed in renal tubular epithelial cells in Sham group;In UUO group,the positive expression of renal interstitium and tubular epithelial cells were obviously increased;While in EPL and SLT groups,the number of possitive expression cells was more than that in sham group but less than that in UUO group.The results of SGK-1?TGF-?₁ showed that,in Sham group,they were weakly expressed,mainly in renal tubular epithelial cells;In UUO group,the expression were enhanced,especially in renal tubular epithelial cells and interstitial;While the expression of SGK-1?TGF-?₁ were obviously reduced in both EPL and SLT groups than UUO group.3 The expression of PCNA?SGK-1?TGF-?₁ by Western blotThe result of PCNA showed that,compared with Sham group,it was obviously up-regulated in UUO group,and the difference was statistically significant?P<0.01?;While in EPL and SLT groups,it expressed more than that in Sham group,but less than that in UUO group,and the difference had statistically significances?P < 0.01?.There was no significant difference between EPL group and SLT group?P>0.05?.The result of SGK-1 and TGF-?₁ showed that,compared with Sham group,they were up-regulated obviously in UUO group,and the difference was statistically significant?P<0.01?.While in EPL and SLT groups,SGK-1 and TGF-?₁ expressed more than that in Sham group,but less than that in UUO group,and the difference had statistically significances?P<0.01?.There was no significant difference between EPL group and SLT group?P>0.05?.Conclusions:1 The activation of aldosterone induced by obstructive nephropathy can stimulate the renal cell proliferation,thus taking part in the formation of renal interstitial fibrosis.2 Eplerenone can reduce the kidney injury in obstructive nephropathy by blocking the activation of aldosterone.3 Shenluotong decoction plays a role in down-regulating the expression of SGK-1 and TGF-?₁,so as to inhibit renal cell proliferation and retard renal interstitial fibrosis.