摘要
目的观察免疫抑制剂环孢素A(CsA)对大鼠肾脏成纤维细胞(NRK)增生及细胞因子分泌的影响以及罗格列酮的干预作用,初步探讨罗格列酮对环孢素A肾毒性的保护作用。方法体外培养大鼠肾脏成纤维细胞。四甲基偶氮唑盐(MTT)比色法测定CsA及罗格列酮对细胞增生的影响。RT-PCR检测过氧化物酶体增殖物激活受体(PPAR)γ、TGF—β1mRNA水平。western印迹检测PPARγ、AT1受体、磷酸化细胞外信号调节激酶(p-ERK)及FN蛋白表达。酶联免疫吸附(ELISA)测定细胞培养上清液中TGF-β1的分泌。结果CsA可明显抑制NRK细胞增生,且呈剂量和时间依赖性(P<0.05)。罗格列酮与CsA合用后,对细胞增殖的抑制作用更明显(P<0.01)。CsA刺激NRK细胞PPARγ、TGF—β1、AT1受体及FN的产生(P<0.05),罗格列酮可下调这些改变(P<0.05)。结论罗格列酮可减轻CsA所致的NRK细胞毒性。
Objective To investigate the protective effects of rosiglitazone (RSG) on normal rat kidney cells (NRK) damaged by cyclosporine A(CsA). Methods The NRK cells were cultured with CsA or with CsA plus rosiglitazone. The cellular proliferation was determined by MTT colorimetry. The mRNA expression of TGF-β1 and PPARγ was detected by RT-RCR. Protein levels of PPARγ, p-ERK,FN and AT1R were examined by Western blotting. The level of TGF-β1 in the supernatants was measured by enzyme-linked immunosorbant assay (ELISA). Results CsA inhibited the proliferation of NRK cells in dose and time dependent manner (P < 0.05), treatment with RSG accelerated the suppression of NRK resulted from CsA (P < 0.05). CsA stimulated the expression of PPARγ, TGF-β1, p-ERK, FN and AT1R compared with control group (P < 0.05), and such up-regulating expression could be decreased by rosilitazone (P<0.05). Conclusion Rosiglitazone may relieve the nephrotoxicily on NRK cells resulted from CsA.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2005年第5期282-285,共4页
Chinese Journal of Nephrology
