| As active substance of mycetogenetic circular peptide 11, cyclosporine A(CsA) had become to be first-line drug to treat rejection reaction after organ transplantation and autoimmune diseases, quickly put into clinical use and then achieved exact therapeutic effects since it was given approval to be immunosuppressive agent for organ transplantation by FDA in 1983. However, the adverse effect especially chronic nephrotoxicity limited its use. Chronic CsA nephrotoxicity(CCN) is characterized by progressive renal dysfunction,afferent arteriolopathy,inflammatory cell influx,striped tubulointerstitial fibrosis,and increased intrarenal immunogenecity. Mihatsch etc. analyzed kidney biopsy data of kidney transplantation patients, verified that incidence of kidney tubules poisoning due to CsA was 9%-37%, incidence of acute kidney function failure accompanied with diffuse interstitial fibrosis was 0%-19%, while incidence of stripe-like interstitial fibrosis and CsA-related arterial diseases were 5%-50% and 5%-30% respectively. The exact mechanism of this complication is not well understood,and multiple factors are implicated. The pathogenesis of CCN was considered to be relevant with glomerulus ischemia, rennin angiotensin system (RAS) activation, apoptosis, endoplasmic reticulum stress, autophagy, mononuclear macrophage(MC/MP) infiltration and inflammation, regulation disbalance of TGF-β1 and MMPs/TIMPs, osteopontin, nuclear factor-kappa B (NF-κB), Toll-like receptor and so on. Focus on its pathogenesis, a lot of researches had tried to intervent and treat CCN by using western durgs, Chinese drugs as well as reduction or even interruption of CsA.As one of traditional Chinese drugs, Rhubarb's effective parts (Rep) is eluate produced by rhababerone extraction passing through macroporous adsorptive resin which includes five ingredients, emodin, chrysophanol, emodin monomethyl ether, rhein and aloe-emodin. Satisfactory therapeutic effects of Rep were reported for improving cerebral ischemia, cerebral metabolism and resisting rejection reaction. Another drug—Sulodexide is a kind of highly purified biological product. It belongs to glycosaminoglycans(GAGs) drugs with biological activity of protecting kidney including heparin, low molecular heparin, heparin sulphate, sulphuric dematan and glycosaminoglycan mixture. Sulodexide consists of two kinds of GAG with the activity of preventing diabetic nephropathy(DN) in experimental models. It was found to resist the overexpression of TGF-β1 and substrate synthesis induced by high-concentration glucose in kidney glomerulus cells, and repair disfunction of endothelial cell during the study of DN. Although both Rep and Sulodexide had the prevention activity for kidney, no document reported the therapeutic effect of two drugs to treat CCN. So we established CCN rat model, using Sulodexide and Rep to treat CCN at different timepoint and different dosage and explore the protective and therapeutic mechanism meanwhile.The article was divided into two parts.Part 1 Intervention effect of Rep and Sulodexide on CCN ratsObjective:To observe whether Rep and Sulodexide have protective role for CCN and to explore the fundamental mechanism. Methods:1. Experimental grouping and biochemical indicator determination:54 SPF healthy male Wistar rats (180～200g) were divided into 9 groups randomly after 1 week's adaptive feeding with low salt diet and free drinking:Group 1, normal control group (VH3:olive oil 5ml·kg-1·d-1×28 d); Group 2, model group (MX3:CsA 25mg·kg-1·d-1×28 d); Group 3, positive control group (YDY:CsA 25mg·kg-1·d-1+ benazepril 5mg·kg-1·d-1×28 d, the two drugs administered over 2 hours intervals); Group 4, Rep intervention group 1 (DY1:after Rep 50mg·kg-1·d-1×7d, CsA 25mg·kg-1·d-1+Rep 50mg·kg-1·d-1×28d, the two drugs administered over 2 hours intervals); Group 5, Rep intervention group 2 (DY2:CsA 25mg·kg-1·d-1+Rep 50mg·kg-1·d-1×28 d, the two drugs administered over 2 hours intervals); Group 6, Rep intervention group 3 (DY3:after CsA 25mg·kg-1·d-1×7d,CsA 25mg·kg-1·d-1+ Rep 50mg·kg-1·d-1×21d, the two drugs administered over 2 hours intervals); Group 7, Sulodexide intervention group 1 (WY1:after Sulodexide 10mg·kg-1·d-1×7d, CsA 25mg·kg-1·d-1+Sulodexide 10mg·kg-1·d-1×28d); Group 8, Sulodexide intervention group 2(WY2:CsA 25mg·kg-1·d-1+Sulodexide 10mg·kg-1·d-1×28d); Group 9, Sulodexide intervention group 3 (WY3:after CsA 25mg·kg-1·d-1×7d, CsA 25mg·kg-1·d-1+Sulodexide 10mg·kg-1·d-1×21d). Based on experimental design, CsA and Rep were taken by intragastric administration, Sulodexide was given by intraperitoneal injection.All rats were killed on day 28th. During the experimental period, rats in each group were raised freely. Rat's body weight was measured at the last day in every week before and after medication, intragastric administration and abdominal injection dosage would be adjusted. At the same time, venous blood from rat tail was collected to measure serum creatinine(Scr) and blood urea nitrogen(BUN).2. Harvested kidney tissues were fixed in periodate-lysine-paraformaldehyde solution and embedded in wax.After dewaxing,2-μm sections were processed and stained with HE,PAS and Masson. Pathological changes,hyaline degeneration of small arteries and renal interstitial fibrosis(RIF) were observed. A semiquantitative score was used to assess the extend of changes in each category.Hyalinosis of small arteries was determined by counting afferent and the juxtaglomerular arterioles available for examination with a minimum of 50 glomeruli per biopsy assessed:0=no hyalinosis; 1 =early hyaline changes; 2=pronounced hyalinosis.RIF was estimated by counting the percentage of injured areas per field and scored as follows:0=normal interstitium,0.5 =<5% of interstitium areas injured,1-5% to 15%,1.5=16% to 25%,2=26% to 35%, 2.5=36% to 45%, and 3=>45%.3. The ultrastructural changes of kidney tissues were also observed by electron microscope.4. Kidney was isolated from rat on ice and the following indicators of the kidney tissues were examined.4.1 Immunohistochemical detection:CD68, collagenⅢand collagenⅣexpression;4.2 Reverse transcription-polymerase chain reaction (RT-PCR) assay:TGF-β1 and TIMP-1 mRNA expression;4.3 ELISA detection:TGF-β1, TIMP-1 and MMP-9 protein expression.Results:1.Body weight, BUN, Scr1.1 Body weight:1 week after CsA administration, the rat in MX3 group growed delay, the weight decreased after 2 weeks vs VH3; 2-4 weeks,the rat weight of YDY and all intervention groups was higher than that of MX3 group (P<0.05), DY3, WY3, YDY group lower than VH3 group(P<0.05), other group was similar to VH3 group (P>0.05).1.2 BUN:1 week after CsA administration, the BUN level of MX3 group began to increase,after 2 weeks,it was higher (P<0.05) than VH3 group; two weeks later the BUN level of DY1, WY1, WY2 group didn't increase (P>0.05), lower than MX3 group(P<0.05), similar to VH3 group(P> 0.05).1 week after, the BUN level of YDY, DY3, WY3 increased apparently,after 3 weeks it began to decline, but higher than VH3 lower than MX3 group(P<0.05);however,the rat BUN level of DY2 group was lower than the above-mentioned three groups (P<0.05),it was similar among the three groups (P> 0.05).1.3 Scr:After 1 week, the rat Scr level of MX3 group increased (P<0.05) vs VH3. At 2-4 weeks that of YDY and all intervention groups was lower than MX3 group.Compared with VH3 group, DY1 group decreased (P<0.05), DY2, WY1 group no changes (P>0.05), YDY and other intervention groups increased (P<0.05). The Scr level of WY2 was lower than YDY, DY3, WY3 group(P<0.05).The Scr level of DY3, WY3 at 4 weeks decreased (P<0.05) and lower than YDY group (P<0.05).2. Light microscopy:In normal control group, light microscopy showed normal structure of kidney tissues. In MX3 group, capillary loop opened badly in kidney glomerulus, cellular and intercellular vacuolization,tubular collapse and dilation.Striped interstistial fibrosis was present with thickening of the tubular basement membranes and Bowman's capsule.Muscular thickening of intrarenal arteriolar vessel wall,arterilar hyalinosis and injury appeared, and focal glomerulosclerosis was found occasionally.The above-mentioned pathological changes of YDY and intervention groups significantly reduced. The hyaline degeneration and RIF of erery group were as follows.2.1 Hyalinosis of small arteries score:The score was higher in YDY and intervention groups than VH3 group,lower than MX3 group (P<0.05), higher in DY1, DY2, DY3, WY3 group than YDY group (P<0.05). Rep vs Sulodexide:The score in Sulodexide groups in various time points was lower than Rep groups(P<0.05), and increased at the third time point as well as higher than at the previous two time points (P<0.05), no difference was found between the first two time points (P>0.05).There was no difference among the various time points in Rep groups(P>0.05).2.2 RIF score:The score in YDY and intervention groups was higher than VH3 group (P<0.05), lower than MX3 group (P<0.05), in which the score of DY3, WY3 group was higher than YDY group (P<0.05), similar in the other intervention groups with YDY group (P>0.05). Rep vs Sulodexide:At the first time point it was lower in Sulodexide group than Rep group (P<0.05), no difference was found between the two medicine groups at the other two time points (P>0.05). In both medicine groups it was higher at the third time point than that at the front two time points (P<0.05), no difference was found between the front two time points (P>0.05).3. Renal ultrastructure changes:The structure was normal in normal control groups.In rat kidney tissues of MX3 group, glomerular basement membrane thickened, mesangium matrix broadened and increased, podocyte foot process fused extensively and microvillus became sparsed; tubular basement membrane thickened irregularly and broke, epithelial cell appeared a lot of vacuoles, nucleus edge changed to be irregular,chromatin thickened and concentrated, pyknosis,tubular microvillus exfoliated, epithelial cell endolysosome and mitochondria increased significantly, collagen fiber increased in interstitium. The rat lesions of DY1,DY2,WY1,WY2 groups significantly lightened:podocyte foot processes fused occasionly; segmental mesangial matrix mild increased; tubular showed a little bubble, lysosome, mitochondria and the basement membrane intact.Pathological changes of YDY,DY3,WY3 group deteriorated but improved much compared with MX3 group.4. Kidney immunohistochemistry:CD68, collagenⅢand collagenⅣexpression levels in every group showed a similar trend with the inflammatory cell infiltration in HE staining. After 4 weeks, those were higher in model group than in control and intervention groups.4.1 CD68 expression:The expression level was similar in DY1, DY2, WY1, WY2 with VH3 group(P>0.05), lower in YDY, DY3, WY3 than VH3 group (P<0.05).The level was lower in YDY and all intervention groups than MX3 group (P<0.05); it was similar in YDY with DY3, WY3 group (P>0.05), but higher than that in the other intervention groups (P<0.05). Rep vs Sulodexide:At the third time point in Rep group it was lower than Sulodexide group (P<0.05), there was no difference between the two medicine groups at the other two time points (P>0.05). Both medicines showed higher level at the third time point than the front two time points (P<0.05), no difference between the front two time points (P>0.05).4.2 CollagenⅢexpression:Compared with VH3 group, collagenⅢexpression level was similar in DY2, WY2 groups (P>0.05), higher in YDY and the other intervention groups (P<0.05). Compared with MX3 group,it was lower in YDY and all intervention groups (P<0.05). While compared with YDY group, the level in WY1 group had no change (P>0.05), DY3, WY3 group increased (P<0.05), other intervention groups decreased (P<0.05). Rep vs Sulodexide:At the third time point it was lower in Sulodexide than Rep group(P<0.05), there was no difference between the two medicine groups at the other two time points (P>0.05). Both medicines showed it was higher at the third time point than the front two time points (P<0.05), and higher at the first time point than the second time point (P<0.05).4.3 CollagenⅣexpression:Vs VH3 group, collagenⅣexpression level in WY1 group had no change(P>0.05), the other intervention groups increased (P<0.05). Compared with MX3 group, the level of YDY and all intervention groups decreased(P <0.05); compared with YDY group,the expression level in WY1 group decreased (P <0.05), that in DY3, WY3 group increased(P<0.05), other intervention groups had no difference (P>0.05). Rep vs Sulodexide:The level was lower in Sulodexide group at the first and third time point than Rep group (P<0.05), there was no difference between the two medicine groups at the second time point (P>0.05). Both medicines showed higher at the third time point than the front two time points (P<0.05), in Sulodexide group the level was higher at the second time point than the first one (P <0.05), and in Rep group there was no difference between the front two time points (P >0.05).5. TGF-β1 and TIMP-1mRNA expression5.1 TGF-β1mRNA expression:In VH3 group, there was only a small amount of TGF-β1mRNA expression, it increased significantly in model,YDY and other intervention groups except WY1 group (P<0.05), the expression of rats in YDY and all intervention groups was lower than MX3 group (P<0.05), that was lower in Rep and Sulodexide groups than YDY group (P<0.05). Rep vs Sulodexide:Compared with Rep groups, it decreased in Sulodexide group (P<0.05), and the expression level in Rep groups was 3>2>1(P<0.05).There was no difference between the first and second time point(P>0.05) in Sulodexide groups,but they were all lower than at the third time point (P<0.05).5.2 TIMP-1 mRNA expression:In VH3 group, there was only a small amount of expression, it increased in MX3,YDY and intervention groups(P<0.05), the expression of rats in YDY and all intervention groups was lower than in MX3 group (P<0.05), lower in Rep and Sulodexide groups than in YDY group(P<0.05). Rep vs Sulodexide: The expression level was lower in Sulodexide groups at any time point than in Rep groups(P<0.05), while both medicine groups showed gradually increase with the time extending (P<0.05).6. TGF-β1 TIMP-1 and MMP-9 protein expression6.1 TGF-β1 protein expression:Compared with VH3 group,the expression was similar in WY1, WY2 group (P>0.05), decreased in DY1, DY2, WY3 group (P<0.05), increased in DY3, YDY group (P<0.05). The level in each other group was higher than that in MX3 group(P<0.05), lower than VH3 group(P<0.05). Rep vs Sulodexide: There was significant difference among all groups (P<0.05), at the front two time points the expression level in Rep was lower than that in Sulodexide group, and at the third time point, just the opposite.It was higher at the third time point than the other two time points(P<0.05) in Rep groups,no difference between the two ones (P>0.05); and the expression level in Sulodexide groups was similar at the first and second time point(P>0.05), but both higher than the third one (P<0.05).6.2 TIMP-1 protein expression:Compared with VH3 group,the expression level was similar in DY3, WY1, WY2 group (P>0.05); decreased in DY1,DY2, WY3 group (P<0.05); increased in YDY group (P<0.05).It was lower in all the other groups except DY3 group (P=0.05) compared with MX3 group (P<0.05). The TIMP-1 protein expression level in each other group was lower than YDY group (P<0.05). Rep vs Sulodexide:At the front two time points the expression in Rep decreased compared with that in Sulodexide group(P<0.05), but at the third time point,it decreased in Sulodexide compared with that in Rep group(P<0.05).It was higher at the third time point than the other two time points in sulodexide groups(P<0.05); however, lower at the third time point than the other two time points in Rep groups(P<0.05).There was no difference between the front two ones (P>0.05) both in Rep and Sulodexide groups.6.3 MMP-9 protein expression:Compared with VH3 group, no change was found in MX3, WY1 group (P>0.05), the expression increased in YDY group and other intervention groups (P<0.05), among those groups the expression of MMP-9 protein in YDY increased the highest (P<0.05).Rep vs Sulodexide:At the first and third time point the expression level in Sulodexide group was lower than in Rep group (P<0.05), but at the second time point,it was lower in Rep than in Sulodexide group.There was no difference among the three time points (P>0.05) in Rep groups,but the level in Sulodexide groups was 2>3>1(P<0.05).Conclusion:Cyclosporine nephrotoxicity could be significantly reduced,even some toxicity can be eliminated by Sulodexide or Rep intervention. Early intervention,better result would be obtained.The effect of Rep and Sulodexide was better than benazepril in decreasing CsA nephrotoxicity.The comparison between Sulodexide and Rep groups found that administration at the first time point,the effect of Sulodexide was better than that of Rep;however,the effect of two medicines administration at the other two time points matched.Part 2 The therapeutic use of Rep and Sulodexide in CCN ratsObjective:To establish CCN model and investigate whether nephrotoxicity could be eliminated by itself 2 weeks after CsA withdrawal; To observe whether Rep or Sulodexide has the therapeutic effect on CCN simultaneously and to evaluate the curative effect.Methods:1. Experimental grouping:78 SPF healthy male Wistar rats (180-200g) were divided into 9 groups randomly after 1 week's adaptive feeding with low salt diet and free drinking:Group 1, normal control group (VH1:n=6, olive oil 5ml·kg-1·d-1×14d and sacrificed at 14 d; VH2:n=6, olive oil 5ml·kg-1·d-1×14d, later water for injection 5ml·kg-1·d-1×14d and sacrificed at 28 d; VH3:n=6, olive oil 5ml·kg-1·d-1×28 d and sacrificed at 28 d); Group 2, model group (MX1:n=6, CsA 25mg·kg-1×14d and sacrificed at 14 d; MX2:n=6, CsA 25mg·kg-1·d-1×14d, later water for injection 5ml·kg-1·d-1×14 d and sacrificed at 28 d; MX3:n=6, CsA 25mg·kg-1·d-1×28 d and sacrificed at 28 d); Group 3, positive control group (YDZ CsA 25mg·kg-1·d-1×14d, later benazepril 5mg·kg-1·d-1×14d); Group 4:Rep therapy group 1(DZ1:CsA 25mg·kg-1·d-1×14d, later Rep 25mg·kg-1·d-1×14d); Group 5, Rep therapy group 2 (DZ2:CsA 25mg·kg-1·d-1×14d, later Rep 50mg·kg-1·d-1×14 d); Group 6, Rep therapy group 3(DZ3:CsA 25mg·kg-1·d-1×14d, later Rep 100mg·kg-1·d-1×14d); Group 7, Sulodexide therapy group 1 (WZ1:CsA 25mg·kg-1·d-1×14d, later Sulodexide 10mg·kg-1·d-1×14d); Group 8, Sulodexide therapy group 2 (WZ2:CsA 25mg·kg-1·d-1×14d, later Sulodexide 20 mg·kg-1·d-1×14 d); Group 9, Sulodexide therapy group 3 (WZ3:CsA 25mg·kg-1·d-1×14d, later Sulodexide 40 mg·kg-1·d-1×14d). Based on experimental design, CsA and Rep were taken by intragastric administration, Sulodexide was given by intraperitoneal injection. The rats in group 3-8 were all killed at 28d.Group 9 failed to finish the experiment because of two rats were dead.2. Rats observation and all indexes checking methods were as same as Part 1.Results:1.Body weight,BUN and Scr1.1 Body weight:Except group 9, rats in the other groups survived and were included in the last statistics. At the first day, there were no significant differences of rat's body weights among various groups(P>0.05); At 14d, rats'body weights among MX groups,YDZ and therapy groups had no difference(P>0.05), while body weights in above-mentioned groups decreased compared to those in VH group(P<0.05). At 21d, rats'body weights continued to decline in MX3 group, but increased in other groups(except VH1, MX1 group because rats had been sacrificed). To 28d, all rats' body weights were improved. Compared with the results at 21d, body weights in MX3 and WZ2 groups had no significant differences, but those in other groups improved(P <0.05). At 28d, body weights in VH2 and VH3 group increased to the highest level. The difference among MX3 and the other groups was significant (P<0.05). Body weights in all therapy groups had no significance compared with those in YDZ and MX2 groups(P>0.05).1.2 BUN,Scr:At the first day, there were no differences of BUN, Scr among all groups(P>0.05); At 7 d and 14 d, level of BUN, Scr in all MX groups,YDZ and therapy groups increased compared to those of normal control group(P<0.05); At 28 d, level of BUN, Scr in the experimental groups except VH1 and MX1 group(rats had been sacrificed) decreased compared with those in MX3 group(P<0.05); Compared to MX2 group,BUN results in DZ1, DZ2 groups and Scr results in DZ1, DZ2,WZ2 groups increased, while other therapy and YDZ group had no changes(P>0.05). Compared to YDZ group, BUN level in DZ1 and DZ2 groups and Scr level in DZ1, DZ2 and WZ2 groups increased (P<0.05), while other therapy groups had no changes(P>0.05).2. Light microscopy results:In normal control group, light microscopy showed normal structure of kidney tissues. The pathology changes of MX3 group were as same as those in Part1. In MX1 group, the results showed the capillary loop opened badly, monocyte infiltration in interstitial tissue, interspersed striped interstitial fibrosis, partial afferent arterioles hyalinization. However, the above-mentioned pathological changes in MX2 and YDZ group improved significantly, and nearly restored to normal appearance in therapy groups. Hyalinosis and RIF in each group were as follows:2.1 Hyalinosis of small arteries score:No obvious hyalinosis was found in therapy and YDZ groups compared to normal control and MX2 groups (P>0.05), while the hyalinosis was relieved compared to MX1, MX3 groups(P<0.05); the results in therapy groups had no differences with the generation in YDZ group(P>0.05).2.2 RIF score:Slight RIF could be seen in normal control group; model groups(MX1, MX2 and MX3) showed significantly aggravation which most serious RIF in MX3, slightest in MX2 and moderate in MX1 group(P<0.05).The serious degree of RIF was similar in YDZ and MX2 groups(P>0.05). RIF in therapy groups was less serious than YDZ and MX2 group(P<0.05) while more serious than normal control groups(VH1, VH2 and VH3) (P<0.05). Rep vs Sulodexide:RIF in WZ1 group was slightest, and RIF in DZ1, DZ3 and WZ2 was most serious and RIF degree in DZ2 was moderate (P<0.05) in which there were no statistical differences among DZ1, DZ3 and WZ2 groups(P>0.05).3. Ultrastructure changes in kidney tissues:The structure was normal in normal control groups.The changes of MX3 group were as same as those in Part1.The changes in MX1 group showed slighter, podocyte foot process fused partially, mesangium matrix increased slightly, vacuoles, lysosome and mitochondria could be found in tubule, thickness of tubular basement membrane was discrepant, segment broke, nuclear chromatin concentrated and interstitial collagen fiber appeared occasionally. Pathological changes in MX2 group improved obviously compared to the results in MX1 and MX3 groups, and the pathology was nearly normal in YDZ and therapy groups.4.Immunohistochemistry results in kidney tissues4.1 CD68 expression:The level of CD68 expression from low to high was VH1, VH2,VH30.05). Rep vs Sulodexide:Results in DZ1 group were lowest, and subsequently increased in DZ2, WZ1, WZ2, DZ3 group in turn(P<0.05, except the difference between those of WZ2 and DZ3 group).4.2 CollagenⅢexpression:CollagenⅢexpression level was lowest in normal control group (VH1, VH2 and VH3 group), DZ1, DZ2 and DZ3 group(P>0.05). The results in model groups increased (MX3>MX2>MX1,P<0.05).Changes in YDZ, WZ1 and WZ2 groups were similar to those of MX2 group(P>0.05), while higher than MX1 group(P<0.05) and lower than MX3 group(P<0.05). Rep vs Sulodexide:Level in Rep therapy groups was lower than Sulodexide therapy groups(P<0.05), while no difference was found in different dosage groups using the same therapeutic drug.4.3 CollagenⅣexpression:There were no statistical differences among collagenⅣexpression level in normal control group(VH1, VH2 and VH3 group), MX2, DZ2 and WZ1 group(P>0.05); Level in MX1, MX3, YDZ, DZ1, DZ3 and WZ2 increased (P<0.05) which was highest in MX3 group(P<0.05),higher in YDZ group(P<0.05) and the lowest in the other four groups(P<0.05, level in the four groups was similar, P >0.05). Rep vs Sulodexide:Results in DZ2, WZ1 were comparable low(level in the two groups was similar, P>0.05), and subsequently increased in DZ3, WZ2 and DZ1 group in turn (P<0.05, level of the above three groups was similar, P>0.05).5. TGF-β1 and TIMP-1mRNA expression5.1 TGF-β1mRNA expression:TGF-β1mRNA expression level was lowest in DZ2 group, and subsequently increased in VH1, VH2, VH3, DZ1, DZ3 and WZ1 group in turn, and the level was even higher in WZ2, YDZ, MX2, MX1 group, and level of MX3 group was highest(P<0.05). Rep vs Sulodexide:Results in DZ2, WZ1 and DZ1 group were lower(level in the three groups was similar, P>0.05), level in DZ3 and WZ2 groups (level in the two groups was similar, P>0.05) increased compared to the above-mentioned three groups(P<0.05).5.2 TIMP-1mRNA expression:TIMP-1mRNA expression level was lowest in WZ1 group, lower in WZ2 group, and subsequently in VH1, VH2, VH3 and DZ1 group, then increased in MX2, DZ2, YDZ, DZ3, MX1 group in turn, level in MX3 group was highest(P<0.05). Rep vs Sulodexide:The group sequence according to TIMP-1mRNA expression level from low to high was WZ1, WZ2, DZ1, DZ2 and DZ3(P<0.05), and level in Sulodexide therapy groups was all lower than that in Rep therapy groups(P<0.05).6. TGF-β1,TIMP-1 and MMP-9 protein Expression6.1 TGF-β1 protein expression:TGF-β1 protein expression restored to normal level in MX2 after CsA withdrawal and increased in YDZ and all drug therapy groups(P<0.05), results in WZ1, DZ2 and DZ1 group were higher than those of YDZ group(P<0.05), while level in WZ2 and DZ3 was similar to that of YDZ group(P >0.05). Rep vs Sulodexide:No difference was found among DZ3, WZ1 and WZ2 groups(P>0.05), the differences among the other groups had statistical significance(P <0.05) which level in WZ2 and DZ3 groups lowest, and subsequently WZ1, DZ2 and DZ1 in turn.6.2 TIMP-1 protein expression:TIMP-1 protein expression restored to normal level in MX2 after CsA withdrawal, and increased in drug therapy groups (P<0.05) except benazepril group. Rep vs Sulodexide:Level in DZ3, WZ1 and WZ2 was lowest(three groups'results were similar, P>0.05), lower in DZ2 group and highest in DZ1 group(P<0.05).6.3 MMP-9 protein expression:MMP-9 protein expression was highest in WZ2 group, lower in MX1, DZ1, DZ2 and WZ1 group (level in these groups was similar, P >0.05), lowest in VH3, MX2, MX3, DZ3, VH1, VH2 and YDZ(P<0.05) which level in these groups was similar(P>0.05). MMP-9 protein expression restored to normal level in MX2 after CsA withdrawal(compared to normal control group, P>0.05). Results in therapeutic groups except DZ3 and YDZ group increased compared to normal control group and MX2, MX3, MX1 group(P<0.05). Rep vs Sulodexide:Level in WZ2 group was highest, that in DZ1, DZ2 and WZ1 was comparable higher while level in DZ3 group was lowest. There was difference between DZ3 and DZ1, DZ2, WZ2 groups (P<0.05),while the differences among level in other groups had no statistical significance(P>0.05). Conclusions:After CsA withdrawal,common indexes such as body weight,BUN,Scr and small artery hyalinosis of rat kidney tissues could be restored, RIF could be improved slightly,but the administration of Sulodexide or Rep could improved RIF obviously.When 10 mg·kg-1·d-1 Sulodexide or 50mg·kg-1·d-1 Rep was administered,the best result appeared.The two medcines'effect matched each other.
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