| [Background and Objective]Cyclosporin A(CsA) is the most important first-line drug for the treatment of rejection after transplantation. Side effects of CsA are gradually discovered while the drugs are generalized in transplantation. Nephrotoxicity of CsA is the most urgent problem to be resolved. Fluvastatin belongs to 3-hydromxy-3-methylgutaryl coenzyme A reductase inhibitors the most classical and effective drug to improve hyperlipemia. Statins have been widely used in the treatment of kidney diseases. Recently, statins are found that not only can lower the levels of cholesterol and LDL in serum but also have nephroprotective effects which is independent of their lipid-lowering properties. Because dyslipoproteinemia always occur in patients who have taken CsA, usage of statins not only can improve the dyslipoproteinemia resulted from CsA but also probably have nephroprotective effects on chronic CsA toxicity which is independent of their lipid-lowering properties, there are few reports about this topic up to now ,however. To eliminate the complexity of amimal experiment and the effect of statins to the hyperlipemia, we culture Normal Rat Kidney (NRK) cells in vitro. NRK cells are stimulated with CsA to imitate a humanmodel of CsA nephrotoxicity, then we observe the effects of fluvastatin and CsA on the NRK cells, and study the possible mechanism of the nephroprotective effects of fluvastatin on NRK cells.[Methods]NRK cells were cultured in routine ways, when cells covered all over the bottom of the culture bottles, they were used for the experiments. (1) Detecting cell proliferations with MTT colorimetry. Cultured NRK cells with CsA, the cells were devided into four groups: control group, CsA0.5μg/ml group,CsA l.0μg/ml group and CsA 2.0μg/ml group, collected cells of every group at 24 48 and 72h. The appropriate concentration was l.0μg/ml and the appropriate hour was 24 when NRK cells were stimulated with CsA based on the results of MTT colorimetry. Cultured NRK cells with CsA and fulvastatin, the cells were devided into five groups: control group, CsA1.0μg/ml group, CsA1.0μg/ml +Flu0.1μmol/L group, CsA1.0μg/ml +Flul.0μmol/L group and CsA1.0μg/ml +Flu10μmol/L group, collected cells of every group at 24h and detected cell proliferations with MTT colorimetry. (2) Detecting the mRNA expression of transforming growth factor- β 1 (TGF- β 1) connective tissue growth factor (CTGF) and c-fos with RT-PCR. Cultured NRK cells with CsA, the cells were devided into four groups: control group, CsA 0.5μg/ml group, CsA l.0μg/ml group and CsA 2.0μg/ml group, collected cells of every group at 24h and detected the mRNA expression of TGF- β 1. CTGF and c-fos with RT-PCR. Cultured NRK cells with CsA and fulvastatin, the cells were devided into four groups: control group, CsA1.0μg/ml group, CsA1.0μg/ml +Flu1.0μmol/L group and CsA1.0μg/ml +Flu10μmol/L group, collected cells of every group at 24h and detected the mRNA expression of TGF- β 1, CTGF and c-fos with RT-PCR. ( 3 ) Detecting the protein level of fibronectin (FN) with Western blotting. The groups were same to RT-PCR, cells of every group were collected at 24h and detected the protein level of FN with Westeren blotting.[Results] 1. Cell proliferations: Compared with control group, cell proliferation of NRK in CsA1.0/ig/ml group were suppressed after being cultured with CsAfor 24h (0.760+0.033 vs 0.870+0.029,P<0.01) . Fluvastatin promoted the suppression of NRK resulted from CsA, and the difference between CsALO/ig/ml + Flu 0.1/anol/L group and CsA 1.0/zg/ml group was significance (0.562+0.068 vs 0.753+0.050, P<0.01) .2. TGF- P 1: The expression of TGF- P 1 mRNA upregulated markedly by CsA dose dependently, the difference between CsA 1.0//g/ml group and control group was significance (0.530+0.041 vs 0.094+0.038,P<0.01) .Fluvastatin down-regulated the expression of TGF- P 1 mRNA significantly, the density value of TGF- P 1 mRNA in CsA1.0//g/ml + Flu 1.0/zmol/L group is 0.411+0.046, significantly lower than that of CsA 1.0/*g/ml group (0.517±0.064,P<0.05) .3. CTGF: The expression of CTGF mRNA upregulated markedly by CsA dose dependently, the difference between CsA l.Qwg/ml group and control group was significance (0.418+0.060 vs 0.095 + 0.030,P<0.05) .Fluvastatin down-regulated the expression of CTGF mRNA significantly, the density value of CTGF mRNA in CsA1.0/*g/ml + Flu l.Qumol/L group is 0.366+0.041 , significantly lower than that of CsA l.Qug/ml group (0.474+0.049, P<0.05) .4. c-fos: The expression of c-fos mRNA upregulated markedly by CsA dose dependently, the difference between CsA l.Qug/ml group and control group was significance (0.462 ±0.021 vs 0.096+ 0.026,P<0.01) .Fluvastatin down-regulated the expression of c-fos mRNA significantly, the density value of c-fos mRNA in CsAl.O^g/ml + Flu 1.0/unol/L group is 0.466+0.028 , significantly lower than that of CsA l.Ojug/ml group (0.556+0.048, P<0.05) .5. FN: The expression of FN protein upregulated markedly by CsA dose dependently, the difference between CsA 1.0/ig/m\ group and control group was significance (0.5 61 +0.053 vs 0. 342+0.024JP<0.05) .Fluvastatin down-regulated the expression of FN protein significantly, the density value of FN protein in CsAl.O/ig/ml + Flu 1.0/imol/L group is 0.456+0.049 , significantly lower than that of CsA l.O^g/ml group (0.547 ± 0.059, iM).05) .[Conclusion] 1. It was demonstrated that fluvastatin suppressed cell proliferation of NRK in vitrowhen cultured with CsA together and this effect was independent of its lipid-lowering properties.2. It was demonstrated that fluvastatin decreaced the expressions of TGF-P In CTGF and FN resulted from CsA, therefore reduced the extracellular matrix accumulation, which might relate to the suppression of c-fos expression resulted from CsA.
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