| The introduction of cyclosporine A into clinical practice has resulted in a major improvement in the short term outcomes of solid organ transplantation and the treatment of autoimmune disease. However, with increased use has come the recognition that CsA has severe deleterious effects on renal structure and function making chronic nephrotoxicity a major limiting side effect. Acute renal dysfunction due to CsA is reversible since it involves renal hemodynamic dysfunction and is not associated with any permanent histology changes. On the other hand, chronic CsA nephrotoxicity may progress to an irreversible renal lesions characterized by striped interstitial fibrosis, tubular atrophy and hyalinosis of the afferent arterioles. Many studys have showed that transforming growth factor-β1 (TGF-β1) is implicated in the fibrosis of chronic CsA nephrotoxicity directly or indirectly and that suppression of over production of TGF-β1 may prevent the accumulation of extracellular matrix in human and experimental glomerular disease. In order to prevent cyclosporin A chronic nephrotoxicity in rats,so firstly we establish a rat model of cyclosporin A chronic nephrotoxicity and observe the expression of TGF-β1 in experimental chronic cyclosporine A nephropathy;secondly, we try introducing phosphorothioate antisense oligodeoxynucleotides for TGF-β1 into the kidney of chronic cyclosporine A nephropathy by cationic liposome-mediated gene transfer method to inhibite the overproduction of TGF-β1,and observe the effect of cationic liposome-mediated phosphorothioate antisense oligodeoxynucleotides for TGF-β1 on chronic nephrotoxicity of cyclosporin A in rats.Methods:First PartUnder the condition of low salt diet ,rats were divided into group A and group B randomly.Rats of group B were treated with CsA(15mg.kg-1.d-1) once a day by intraperitoneal injection for 1 week(B1:n=3),2 weeks(B2:n=3) ,3 weeks(B3:n=6), 4 weeks(B4:n=6) respectively while rats of group A as controls ,were treated with an equiverlent dose of normal saline ,then both of the groups were sacrificed and we observed the changes of histopathologyof kidney and level of serum creatinine in the rats .The expression of TGF-β1 protein and mRNA were also tested by immunohistochemistry and RT-PCR in different groups; Second PartUnder the condition of low salt diet ,rats were divided into group A(n=16) ,group B(n=16), group C(n=16) and group D(n=16) randomly. Rats of group B,C and D were treated with CsA (15mg.kg-1.d-1) once a day by intraperitoneal injection for 3 weeks(n=8),4 weeks(n=8), while rats of group A as controls, were treated with an equiverlent dose of normal saline. After rats had been treated with CsA for 15 days and 20 days, we introdued phosphorothioate antisense oligodeoxynucleotides for TGF-β1 into the kidney of the group C and cationic liposome-mediated phosphorothioate antisense oligodeoxynucleotides for TGF-β1 into the kidney of the group D respectively, then we compared the changes of level of serum creatinine and histopathology of tubulointerstitial fibrosis of kidney between different groups. The expression of TGF-β1 protein and mRNA in kidney of different groups were also tested by immunohistochemistry and RT-PCR respectively. Result:First PartUnder the condition of low salt diet, rats were treated with CsA (15mg.kg-1.d-1) once a day by intraperitoneal injection for 4 weeks, and then increased level of serum creatinine and characteristic histopathology of tubulointerstitial fibrosis which shows similarity to the structural lesions described in patients were observed. The expression of TGF-β1 protein and mRNA was increased significantly in the kidney of rats which had been treated with CsA for 3 weeks(P<0.01),and with the prolonging of administration of CsA,they increased more significantly.Second PartIn the group C, the expression of TGF-β1 protein and mRNA ,compared with that of the group B, has not changed significantly . In the group D, a marked decrease in expression of mRNA was confirmed by RT-PCR(P<0.01). By im... |
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