| Cyclosporine A (CsA) has become the cornerstone of clinical immunosuppressive therapy. In essence, it predominantly improves the short-term survival of organ transplantation. CsA is the first-line drug for the treatment of rejection after organ transplantation, although numerous new ones come into view. Nevertheless, CsA therapy has fierce side effects, especially nephrotoxicity and hepatotoxicity. Fibrosis of liver and kidney induced by CsA is still a problem-in-discussing of the development of organ transplantation. Rosiglitazone (RGZ), a member of thiazolidinediones (TZDs), has universal effects by activating peroxisomeproliferator-activated receptor y (PPAR r). PPAR r, a member of type II nuclear receptor superfamily, not only involves in the cellular metabolism, but also plays an important role on ameliorating inflammation and fibrosis. Therefore, we hypothesize that PPAR y may be a newtarget for the management of conditions induced by CsA. There is no such report up to now, however. We establish chronic CsA nephropathy rat model at the basis of low salt diet (LSD) and culture NRK (normal rat kidney cells) and BRL (Buffalo rat liver cells) as well, to observe the protective effects of RGZ on nephrotoxicity and hepatotoxicity induced by CsA both in vivo and in vitro, and then explore mechanisms of these effects.Part I Experimental Study of Protective Effects of Rosiglitazone on Chronic CsA NephropathyResearch in VivoMethods1. Healthy male SD rats (BW240~260g) fed with LSD one week later were randomly allocated into three groups as described below: (1) control (n=12), LSD; (2) model (n=15), LSD + CsA(15mg. Kg d-1); (3) RGZ treated (n=15), LSD + CsA(15mg. Kg-1 d-1) + RGZ (5mg. Kg-1 d-1). 6 ones of each group were sacrificed randomly 2 weeks later, the rest were sacrificed at the end of the experiment (5 weeks later).2. General and biochemical parameters: body weight was recorded at the beginning and the end of the experiment. Urinary creatinine (uCr), serum creatinine (sCr) and albumin (Alb) were investigated with automatic analyzer. Creatinine clearance (Ccr) is calculated with uCr, sCr and 24h urine volume.3. Histological evaluation: interstitial mononuclear cells were counted in HE staining sections, and interstitial fibrosis was calculated by Masson's staining.4. Immunohistochemical analysis (IHC): transforming growth factor-B1 (TGF-B1),collagen III(Col III), PPAR v and a -smooth muscle actin (a -SMA) were stained with SP kit.5. mRNA expressions of TGF-B1 and PPAR r were detected by Reverse transcription-polymerase chain reaction (RT-RCR).6. Protein expressions of PPAR y , Fibronectin (FN), Angiotensin II type 1 receptor(AT1R) and phosphorylation-extracellular signal regulated kinase (p-ERK) were detected by Western blotting.7. The plasma and tissue activity of Angiotensin II (Ang II) were detected by radioimmunology (RI).Results1. Body weight: there was significant difference of body weight of rats in each group both of 2 and 5 weeks (P<0.05). Weight loss was markedly ameliorated in rats treated with RGZcompared with that in model group at 5 weeks (P<0.05).2. Biochemical parameters: when compared to control group, CsA caused a significant decrease (P<0.05) in level of Ccr and serum Alb level. Treatment with RGZ produced a significant improvement in level of Ccr at 5 weeks (P<0.05), but failed to improve level of Alb(P>.05).3. Interstitial mononuclear cells infiltration: the amount of interstitial mononuclear cells infiltrated in model group increased markedly at 2 weeks (P<0.05), and still patent at 5 weeks (P<0.05) although decreased compared with that at 2 weeks. Increase of interstitial mononuclear cells was significantly ameliorated by RGZ both of 2 and 5 weeks (P<0.05).4. Interstitial fibrosis: CsA treated rats developed a striped pattern of interstitial fibrosis. Treatment with RGZ resulted in significantly less severe interstitial fibrosis compared with model group at 5 weeks (P<0.05). A s...
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