Centro di Ricerca E. Menni, Fondazione Poliambulanza-Istituto Ospedaliero, Brescia, Italy.
Centro di Ricerca E. Menni, Fondazione Poliambulanza-Istituto Ospedaliero, Brescia, Italy; School of Veterinary Science, University of Liverpool, Neston, United Kingdom.
Cytotherapy. 2014 Jan;16(1):17-32. doi: 10.1016/j.jcyt.2013.07.002. Epub 2013 Oct 1.
We recently demonstrated that injection of conditioned medium (CM) generated from cells of the mesenchymal region of human amniotic membrane (AMTCs) reduces bleomycin-induced lung fibrosis in mice, suggesting a crucial role of paracrine factor(s) secreted by AMTCs in these beneficial effects. We further investigated this hypothesis, the mechanisms involved, the effects on some lung functional parameters and whether AMTC-secreted effector(s) are specific to these cells and not produced by other cell types, extending the time of analysis up to 28 days after treatment.
Bleomycin-challenged mice were either treated with AMTC-CM or CM generated from human skin fibroblasts, human peripheral blood mononuclear cells or Jurkat cells, or were left untreated. Mouse lungs were analyzed for content of pro-inflammatory and pro-fibrotic molecules, presence of lymphocytes and macrophages and for fibrosis level (through histological semi-quantitative evaluation and quantitative measurement of collagen content). Arterial blood gas analysis was also performed.
Up to 28 days after delivery, AMTC-CM-treated mice developed reduced lung fibrosis with respect to mice treated with other CM types. AMTC-CM-treated mice had comparatively better preservation of blood gas parameters and showed lower lung content of interleukin-6, tumor necrosis factor-α, macrophage inflammatory protein-1α, monocyte chemoattractant protein-1 and transforming growth factor-β associated with reduced lung macrophage levels.
AMTC-CM prevents lung fibrosis in bleomycin-challenged mice, improving survival and preserving lung functional parameters such as blood gas exchanges. The specificity of AMTC-CM action was indicated by the absence of fibrosis reduction when other CM types were used. Finally, we provide some insights into the possible mechanisms underlying AMTC-CM-mediated control of fibrosis.
我们最近的研究表明，注射来源于人羊膜间充质区细胞的条件培养基（CM）可减少博来霉素诱导的小鼠肺纤维化，这表明 AMTC 分泌的旁分泌因子在这些有益作用中起着关键作用。我们进一步研究了这种假说，涉及的机制，对某些肺功能参数的影响，以及 AMTC 分泌的效应物是否仅针对这些细胞，而不是由其他细胞类型产生，将分析时间延长至治疗后 28 天。
用 AMTC-CM 或从人皮肤成纤维细胞、人外周血单核细胞或 Jurkat 细胞生成的 CM 处理博来霉素处理的小鼠，或不进行处理。分析小鼠肺中促炎和促纤维化分子的含量、淋巴细胞和巨噬细胞的存在以及纤维化程度（通过组织学半定量评估和胶原含量的定量测量）。还进行了动脉血气分析。
在分娩后 28 天内，与用其他 CM 类型处理的小鼠相比，用 AMTC-CM 处理的小鼠肺纤维化程度降低。与用其他 CM 类型处理的小鼠相比，用 AMTC-CM 处理的小鼠血气参数保持较好，肺中白细胞介素 6、肿瘤坏死因子-α、巨噬细胞炎症蛋白-1α、单核细胞趋化蛋白-1 和转化生长因子-β的含量较低，与肺巨噬细胞水平降低有关。
AMTC-CM 可预防博来霉素处理的小鼠肺纤维化，提高存活率并保持肺功能参数，如血气交换。当使用其他 CM 类型时，纤维化减少的缺失表明 AMTC-CM 作用的特异性。最后，我们提供了一些关于 AMTC-CM 介导的纤维化控制的可能机制的见解。
