Rapamycin increases transforming growth factor-beta mRNA expression in immortalized rat proximal renal tubular cells.
Rapamycin increases transforming growth factor-beta mRNA expression in immortalized rat proximal renal tubular cells.
复制标题
雷帕霉素增加永生化大鼠近端肾小管细胞中转化生长因子-β mRNA 的表达。
DOI：
10.1097/00007890-200201270-00033
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发表时间：
2002
期刊：
影响因子：
6.2
通讯作者：
Ingelfinger,JulieR
中科院分区：
文献类型：
--
作者：
Swinford,RitaD;Pascual,Manuel;Diamant,Daniel;Tang,Shiow-Shih;Ingelfinger,JulieR
We read the article by Dodge et al.(1) in which the authors reported that rapamycin increases the production of transforming growth factor (TGF)-β 1 by lymphocytes in vitro and in vivo. This finding is particularly interesting because accumulating evidence suggests that excessive production of TGF-β 1 within the kidney is linked to the development of fibrosis and tubular atrophy characteristic of chronic allograft nephropathy, and also to cyclosporine (CsA)(or tacrolimus) and chronic allograft nephropathy (2–5). Indeed, recent studies have indicated that CsA enhances expression of TGF-β 1 in T lymphocytes, murine renal proximal tubule cells, and fibroblasts (4, 5) Thus, it has become clear that delineating those factors that regulate TGF-β 1 expression, in the presence or absence of calcineurin inhibitors or other new immunosuppressive drugs, are important for organ transplantation management. Rapamycin (RAPA) is currently being used as a new immunosuppressant for the prevention of acute allograft rejection (5–7). Transplant protocols using CsA and RAPA in combination may be associated with increased expression of the potent fibrogenic cytokine TGF-β, resulting in potent immunosuppression but also higher nephrotoxicity (1, 3). Because of these theoretical concerns, we determined TGF-β 1 mRNA expression and analyzed possible additive or antagonistic effects when CsA and RAPA were used in combination in an immortalized rat proximal tubular cell line (IRPTC)(8).IRPTC were cultured either with CsA alone (500, 1000, 2000, and 5000 ng/ml); with RAPA alone (1, 5, 10, and 20 ng/ml); or with CsA (500 and 1000 ng/ml) plus RAPA (10 and 20 ng/ml). IRPTC (passages 8–20) were seeded into culture flasks (75 cm) and grown to confluence in defined serum-free media (8). Cells were cultured at 37 C and 5% CO 2 and Dulbecco’s modified Eagle’s medium, which was changed every 2 days. Confluence was achieved at day 4 or 5 after plating. At confluence, all cultures were changed to serum-free Dulbecco’s modified Eagle’s medium and supplemented with 0.5% heat-inactivated bovine serum albumin, for 24 hr before CsA and RAPA addition. All cell culture experiments were performed in triplicate. Total RNA was extracted from confluent cell cultures using standard methods, and Northern blots were performed as described (9). The intensity and area of TGF-β1 and GAPDH mRNA expression (the latter for normalization of data) were determined using the Desk Scan II (V1. 5.1; Hewlett Packard) and Image (version 1.37; Wayne Rasband, Research Services Branch, NIMH, Bethesda, MD).
