摘要：

Objective To observe the process of renal tubular epithelial cell (TEC) transdifferentiation and the expression of Wilms tumor 1 gene (WT1) and to study the effect of WT1 on TEC transdifferentiation induced by IL-1α. Methods TECs were cultured in vitro and WT1 antibody was induced.Confluent cells were cultured for 5 days in DMEM/F12 media containing IL-la (10 ng/ml) , with or without 10 (μg/ml of mouse anli-WTl neutralizing antibody, and the ones cultured in DMEM/F12 medium only served as control. TEC morphologic and phenotype characteristics were observed by light microscope and electron microscope. The expression of α-smooth actin (α-SMA) and WT1 was detected by RT-PCR on day 1, day 2, and day 3 of cell culture. Results TEC cultured in media with IL-1α (10 ng/ml) for 5 days showed clear morphological and phenotypic changes. Many of them appeared fibroblast-like. Becoming elongated, the transformed cells showed marked hypertrophyand lost their cobblestone growth pattern. Scanning electron microscopy showed loss of apical-basal epithelial polarity and surface microvilli on the transformed cells. TEC cultured in media with (10 μg/ml) for 1 day started to re-express WT1 and α-SMA as well. However, WT1 became undetectable on day 3 in a transient pattern. α-SMA's expression was enhanced in a time-dependent manner. TEC cultured in media with IL-1α (10 ng/ml) in the presence of 10 μ-g/ml of mouse neutralizing anti-WTl antibody had little morphological changes, the WT1 and α-SMA's expression clearly decreased in contrast to TEC cultured in media with IL-1α (10 ng/ml). Conclusions TEC embedded in high concentration of cytokine for long period would induce epithelial-myofibroblast transdifferentiation itself. The transient, re-expression of WT1 could play an important role in epithelial- myofibroblast transdifferentiation. The re-expression of WT1 in adult TEC could be the inner promotor of TEC transdifferentiation.Neutralization of WT1 gene product in vitro could counteract TEC transdifferentiation as well as kidney fibrosis.

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