Watanabe Tomoya, Nishimoto Tetsuya, Mlakar Logan, Heywood Jonathan, Malaab Maya, Hoffman Stanley, Feghali-Bostwick Carol
Division of Rheumatology & Immunology, Department of Medicine, Medical University of South Carolina, Charleston, South Carolina, United States of America.
PLoS One. 2017 Jun 26;12(6):e0179917. doi: 10.1371/journal.pone.0179917. eCollection 2017.
The murine bleomycin (BLM)-induced fibrosis model is the most widely used in systemic sclerosis (SSc) studies. It has been reported that systemic delivery of BLM via continuous diffusion from subcutaneously implanted osmotic minipumps can cause fibrosis of the skin, lungs, and other internal organs. However, the mouse strain, dosage of BLM, administration period, and additional important features differ from one report to the next. In this study, by employing the pump model in C57BL/6J mice, we show a dose-dependent increase in lung fibrosis by day 28 and a transient increase in dermal thickness. Dermal thickness and the level of collagen in skin treated with high-dose BLM was significantly higher than in skin treated with low dose BLM or vehicle. A reduction in the thickness of the adipose layer was noted in both high and low dose groups at earlier time points suggesting that the loss of the fat layer precedes the onset of fibrosis. High-dose BLM also induced dermal fibrosis and increased expression of fibrosis-associated genes ex vivo in human skin, thus confirming and extending the in vivo findings, and demonstrating that a human organ culture model can be used to assess the effect of BLM on skin. In summary, our findings suggest that the BLM pump model is an attractive model to analyze the underlying mechanisms of fibrosis and test the efficacy of potential therapies. However, the choice of mouse strain, duration of BLM administration and dose must be carefully considered when using this model.
小鼠博来霉素(BLM)诱导的纤维化模型是系统性硬化症(SSc)研究中使用最广泛的模型。据报道,通过皮下植入的渗透微型泵持续扩散进行BLM的全身给药可导致皮肤、肺和其他内部器官的纤维化。然而,不同报告中的小鼠品系、BLM剂量、给药期以及其他重要特征各不相同。在本研究中,通过在C57BL/6J小鼠中采用泵模型,我们发现到第28天时肺纤维化呈剂量依赖性增加,且皮肤厚度有短暂增加。高剂量BLM处理的皮肤的真皮厚度和胶原蛋白水平显著高于低剂量BLM或赋形剂处理的皮肤。在早期时间点,高剂量组和低剂量组均观察到脂肪层厚度减少,这表明脂肪层的丧失先于纤维化的发生。高剂量BLM还在体外诱导人皮肤真皮纤维化并增加纤维化相关基因的表达,从而证实并扩展了体内研究结果,并证明人体器官培养模型可用于评估BLM对皮肤的影响。总之,我们的研究结果表明,BLM泵模型是分析纤维化潜在机制和测试潜在治疗方法疗效的一个有吸引力的模型。然而,使用该模型时必须仔细考虑小鼠品系的选择、BLM给药持续时间和剂量。