1Department of Burns and Plastic Surgery, Nanjing Drum Tower Hospital Clinical College of Traditional Chinese and Western Medicine Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210008, China.
Department of Burns and Plastic Surgery, Nanjing Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medical School, Nanjing, Jiangsu 210008, China.
Curr Mol Med. 2020;20(7):558-571. doi: 10.2174/1566524020666200106095745.
To study the effect of Adipose-derived stem cells (ADSCs) on fibrosis of hypertrophic scar-derived fibroblasts (HSFs) and its concrete mechanism.
ADSCs have been reported to reduce collagen production and fibroblast proliferation in co-culture experiments. Conditioned medium from adipose-derived stem cells (ADSCs-CM) has successfully inhibited fibrosis by decreasing the expression of collagen type І (Col1) and α-smooth muscle actin (α-SMA) in rabbit ear scar models. Hepatocyte growth factor (HGF), the primary growth factor in ADSCs-CM, has been shown to reverse fibrosis in various fibrotic diseases.
To test the hypothesis that ADSCs inhibit fibrosis of HSFs through the secretion of HGF.
HSFs were treated with DMEM containing 0%, 10%, 50% and 100% concentration of ADSCs-CM. The effect of ADSCs-CM on the viability was determined by cell viability assay, and the collagen production in HSFs was examined by Sirius red staining. Expression and secretion of fibrosis and degradation proteins were detected separately. After measuring the concentration of HGF in ADSCs-CM, the same number of HSFs were treated with 50% ADSCs-CM or HGF. HGF activity in ADSCs-CM was neutralized with a goat anti-human HGF antibody.
The results demonstrated that ADSCs-CM dose-dependently decreased cell viability, expression of fibrosis molecules, and tissue inhibitor of metalloproteinases-1 (TIMP-1), and significantly increased matrix metalloproteinase-1 (MMP-1) expression in HSFs. Collagen production and the ratio of collagen type І and type III (Col1/Col3) were also suppressed by ADSCs-CM in a dose-dependent manner. When HSFs were cultured with either 50% ADSCs-CM or HGF (1 ng/ml), a similar trend was observed in gene expression and protein secretion. Adding an HGF antibody to both groups returned protein expression and secretion to basal levels but did not significantly affect the fibrosis factors in the control group.
Our findings revealed that adipose-derived stem cell-secreted HGF effectively inhibits fibrosis-related factors and regulates extracellular matrix (ECM) remodeling in hypertrophic scar fibroblasts.
研究脂肪来源干细胞（ADSCs）对增生性瘢痕衍生成纤维细胞（HSFs）纤维化的影响及其具体机制。
已有报道称，ADSCs 可在共培养实验中减少胶原产生和纤维母细胞增殖。脂肪来源干细胞条件培养基（ADSCs-CM）通过降低兔耳瘢痕模型中胶原 I 型（Col1）和α-平滑肌肌动蛋白（α-SMA）的表达，成功抑制了纤维化。肝细胞生长因子（HGF）是 ADSCs-CM 中的主要生长因子，已被证明可逆转多种纤维化疾病中的纤维化。
通过检测 ADSCs 通过分泌 HGF 抑制 HSFs 纤维化的假说。
用含有 0%、10%、50%和 100%浓度 ADSCs-CM 的 DMEM 处理 HSFs。通过细胞活力测定法测定 ADSCs-CM 对细胞活力的影响，并通过天狼猩红染色检测 HSFs 中的胶原产生。分别检测纤维化和降解蛋白的表达和分泌。测量 ADSCs-CM 中的 HGF 浓度后，用相同数量的 HSFs 处理 50%ADSCs-CM 或 HGF。用山羊抗人 HGF 抗体中和 ADSCs-CM 中的 HGF 活性。
结果表明，ADSCs-CM 呈剂量依赖性降低 HSFs 的细胞活力、纤维化分子表达和组织金属蛋白酶抑制剂-1（TIMP-1），并显著增加基质金属蛋白酶-1（MMP-1）的表达。ADSCs-CM 还呈剂量依赖性抑制胶原产生和胶原 I 型和 III 型（Col1/Col3）的比值。当 HSFs 与 50%ADSCs-CM 或 HGF（1ng/ml）培养时，基因表达和蛋白分泌也呈现相似趋势。在两组中添加 HGF 抗体可使蛋白表达和分泌恢复到基础水平，但对对照组的纤维化因子没有显著影响。
我们的发现表明，脂肪来源干细胞分泌的 HGF 可有效抑制纤维化相关因子并调节增生性瘢痕成纤维细胞细胞外基质（ECM）重塑。
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