Department of Nephrology, Jingan District Central Hospital/Jingan Branch, Huashan Hospital affiliated to Fudan University, Shanghai, 200040, China.
Department of Nephrology, Jingan District Central Hospital/Jingan Branch, Huashan Hospital affiliated to Fudan University, Shanghai, 200040, China.
Biomed Pharmacother. 2018 Mar;99:306-312. doi: 10.1016/j.biopha.2018.01.005.
CCN family, a group of six extracellular matrix-associated proteins, plays an important role in fibrosis. WISP3 has addressed as a pro-fibrotic molecule in the development of human lung fibrosis. However, whether WISP3 involved in the activation and proliferation of renal fibroblast, and ultimately inhibited fibroblast-myofibroblast transdifferentiation remained unknown. Herein, we found that down-regulated WISP3 was involved in the fibrogenesis of rat renal NRK-49F cells induced by transforming growth (TGF-β1), which was further confirmed in a rat renal fibrosis induced by unilateral ureteral obstruction (UUO). In the present study, we aimed to investigate the roles of WISP3 in NRK-49F fibroblast-myofibroblast transdifferentiation, and the underlying mechanism. Results showed that after TGF-β1 treatment, significant increased cell proliferation, and up-regulated expressions of TGF-β1, connective tissue growth factor (CTGF), α-smooth muscle actin (α-SMA), vimentin, as well as increased concentrations of collagen types I (COL I), collagen types III (COL III) and hydroxyproline in cell culture supernatant were observed, demonstrating a successful establishment of fibroblast-myofibroblast transdifferentiation of NRK-49F cells. Besides, siRNA-WISP3 remarkably promoted the fibrogenesis of NRK-49F cells with or without TGF-β1 treatment, and increased mRNA levels of Axin, demonstrating that activating WNT signaling pathway was the underlying mechanism. However, lentivirus-mediated WISP3 overexpression exerted an opposite effect, protecting NRK-49F cells from transdifferentiation, and decreasing mRNA levels of Axin. In conclusion, the WISP3 played an anti-fibrotic role in NRK-49F cells, and WNT signaling pathway was the potential mechanism. WISP3 was an anti-fibrotic factor in fibroblast-myofibroblast transdifferentiation, and may be used as a possible target for prevention and treatment of human renal fibrosis.
CCN 家族是一组六种细胞外基质相关蛋白，在纤维化中发挥重要作用。WISP3 被认为是人类肺纤维化发展中的促纤维化分子。然而，WISP3 是否参与了肾成纤维细胞的激活和增殖，以及最终抑制成纤维细胞-肌成纤维细胞转分化尚不清楚。在此，我们发现下调的 WISP3 参与了转化生长因子（TGF-β1）诱导的大鼠肾 NRK-49F 细胞的纤维化过程，这在单侧输尿管梗阻（UUO）诱导的大鼠肾纤维化中得到了进一步证实。在本研究中，我们旨在研究 WISP3 在 NRK-49F 成纤维细胞-肌成纤维细胞转分化中的作用及其潜在机制。结果表明，TGF-β1 处理后，细胞增殖显著增加，TGF-β1、结缔组织生长因子（CTGF）、α-平滑肌肌动蛋白（α-SMA）、波形蛋白的表达上调，细胞培养上清液中 I 型胶原（COL I）、III 型胶原（COL III）和羟脯氨酸的浓度也增加，表明 NRK-49F 细胞成纤维细胞-肌成纤维细胞转分化成功建立。此外，siRNA-WISP3 明显促进了 NRK-49F 细胞的纤维化，无论是否有 TGF-β1 处理，并且 Axin 的 mRNA 水平增加，表明激活 WNT 信号通路是潜在的机制。然而，慢病毒介导的 WISP3 过表达则产生相反的效果，保护 NRK-49F 细胞免受转分化，并降低 Axin 的 mRNA 水平。总之，WISP3 在 NRK-49F 细胞中发挥抗纤维化作用，WNT 信号通路是潜在的机制。WISP3 是成纤维细胞-肌成纤维细胞转分化中的抗纤维化因子，可能成为预防和治疗人类肾纤维化的潜在靶点。
