Effects Of Chailing Decoction On Oxidative Stress In Chonic Cyclosporine A Nephropathy Rats

Objective: To investigate the mechanism and protection of Chailing Decoction on chronic cyclosporine A nephropathy by observing the infulence of Chailing Decoction on oxidative damage indexes in chronic cyclosporine A nephropathy rats,then to provide the evidence for the clinical application.Methods: Forty Healthy female clean Sprague-Dawley(SD)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,forty rats were randomly divided into four groups: Control group,CsA group,Valsartan group and TCM group.The rat model with CCN were induced by gavage with CsA 30mg·kg-1·d-1 for 28 days.The same dosage of olive oil was given in control group.At the same time,the rats in Valsartan group and TCM group were respectively treated by gavage with valsartan 10mg·kg-1·d-1 and Chailing Decoction 3g·kg-1·d-1 for 28 days,while the rats in the other two groups received physical saline at equal quantity.The urine collected in metabolic cages every weekend was to be used for testing the contents of 8-hydroxy-2'-deoxyguanosine(8-OhdG)and 8-iso-prostaglandin F2?(8-iso-PGF2?)by enzyme-linked immunoassay(ELISA).On the twenty-ninth day,all the rats were weighed one by one.After anesthesia with 10% chloral hydrate,the first thing was to dissociate the arteria femoralis and cut it off to get blood which would go through rapid centrifugation.Conservation of serum samples to be used for biochemical detection of blood urea nitrogen(BUN)and serum creatinine(Scr),thus calculating creatinine clearance rate(Ccr).The contents of GSH?CAT in serum were detected by colorimetry.The second thing is to cut down the left kidney.In order to observe the renal pathological changes,part of the kidney were fixed by 10% paraformaldehyde for HE?Masson staining.Results:1 The general situation in ratsAll the rats survived and were included in the last statistics.The mental state,activity,fur,diet,drinking and urine volume of rats in control group had no obvious changes,while in CsA group,the rats were listless and bovine,the intake of food and water was down-regulated,its fur turned yellow and loose,some even fell off.Compared with CsA group,the general situation of rats in Valsartan group and TCM group were improved.2 The change of pathomorphology in renal tissueHE staining showed that the renal of rats in control group had no abnormal changes,the tubular epithelial cells arranged regularly without swelling and little inflammatory cells can be seen in renal interstitial.In CsA group,a large number of inflammatory cells were infiltrated in renal interstitial,the tubular epithelial cells were denatured,swelling,even necrosis and shed.There were still some inflammatory cells infiltrated in renal interstitial with the tubular epithelial cells swelling in Valsartan group and TCM group,but it was significantly lighter than Cs A group.Masson staining showed that the renal structure of rats in control group was clear to be seen,the tubular basement membrane was complete,a little collagen deposited in renal interstitial interstitium.In CsA group,the renal structure was disordered,tubular basement membrane was thickened and distorted even disrupted,the collagen in tubular interstitial was significantly increased and distributed just like strip-shaped.There were some degree of collagen deposition in Valsartan group and TCM group,but it was obviously reduced compared with CsA group.3 The Result of Renal functionCompared with control group,Scr and BUN were increased visibly while Ccr was significantly decreased in CsA group(P<0.01).Compared with CsA group,the renal function of rats in Valsartan group and TCM group improved markedly,Scr and BUN were obviously lower while Ccr was significantly higher(P<0.01).There was no significant difference between the two treatment groups(P>0.05).4 The results of colorimetryCompared with control group,the level of serum GSH and CAT was significantly decreased in CsA group(P<0.01);Compared with CsA group,the level of serum GSH and CAT in Valsartan group and TCM group was obviously increased(P<0.01,P<0.05).There was no significant difference between the two treatment groups(P>0.05).5 The results of ELISACompared with control group,the contents of urine 8-OhdG and 8-iso-PGF2? were distinctly higher in CsA group(P<0.01);Compared with CsA group,the contents of urine 8-OhdG and 8-iso-PGF2? in Valsartan group and TCM group were significantly lower(P<0.01,P<0.05).There was no obvious difference between the two treatment groups(P>0.05).Conclusions:1 Chailing Decoction could alleviate the renal pathological damage induced by CsA,and improve renal function by ruducing Scr?BUN and increasing Ccr.2 Chailing Decoction played a role in up-regulating the expression of GSH?CAT in serum,so as to impove oxidative damage and protect the renal function by removing the oxygen free radicals in the body.3 Chailing Decoction had the effect of down-regulating the over expression of 8-OhdG ? 8-iso-PGF2? in urine with a certain ability of anti-oxidative damage,thus improving oxidative stress status in CCN rats,inhibiting renal interstitial fibrosis and retarding the progress of chronic cyclosporine A nephrotoxicity.