表达松弛素的腺病毒可减少瘢痕疙瘩成纤维细胞的胶原合成并上调基质金属蛋白酶的表达：体外实验。
Relaxin-expressing adenovirus decreases collagen synthesis and up-regulates matrix metalloproteinase expression in keloid fibroblasts: in vitro experiments.
机构信息
Seoul, Korea From the Institute for Human Tissue Restoration, Department of Plastic and Reconstructive Surgery, and the Department of Dermatology, Yonsei University College of Medicine; and the Department of Bioengineering, College of Engineering, Hanyang University.
出版信息
Plast Reconstr Surg. 2012 Sep;130(3):407e-417e. doi: 10.1097/PRS.0b013e31825dbf56.
BACKGROUND
The hormone relaxin has been shown to affect the extracellular matrix by inhibiting collagen synthesis and expression in fibroblasts stimulated with a profibrotic agent. It also increases matrix metalloproteinase (MMP) expression. To investigate its effect on expression of collagen and MMPs in keloid fibroblasts and human dermal fibroblasts, the authors introduced a relaxin-expressing adenovirus (dE1-RGD/lacZ/RLX) into a human dermal fibroblast cell line and keloid fibroblasts.
METHODS
Both fibroblasts were infected with dE1-RGD/lacZ/RLX or control virus, and protein levels of relaxin and secreted transforming growth factor (TGF)-β1 were assessed by enzyme-linked immunosorbent assay, and mRNA levels of collagen type I, collagen type III, MMP-1, and MMP-3 were assessed by real-time reverse-transcriptase polymerase chain reaction and enzyme-linked immunosorbent assay. Expression of Smad3 and phosphorylated Smad3 was also examined, and relaxin's effect on Smad2/3 complex localization was evaluated.
RESULTS
When human dermal fibroblasts and keloid fibroblasts were transduced with dE1-RGD/lacZ/RLX or dE1-RGD/lacZ (control), mRNA expression of type I and type III collagen was markedly decreased by relaxin regardless of TGF-β (10 ng/ml) treatment. Expression of Smad3 and phosphorylated Smad3 was reduced in keloid fibroblasts and decreased translocation of Smad 2/3 complex from cytosols to the nucleus of the human dermal fibroblasts with TGF-β after dE1-RGD/lacZ/RLX transduction, suggesting that relaxin reduces collagen synthesis by blocking TGF-β signaling. Analyses revealed that MMP-1 and MMP-3 expression were significantly increased in human dermal fibroblasts and keloid fibroblasts after dE1-RGD/lacZ/RLX transduction.
CONCLUSION
These results suggest that the antifibrotic effect of relaxin-expressing adenovirus may have therapeutic effects on keloids.
背景
已经证明，松弛素通过抑制成纤维细胞中胶原蛋白的合成和表达，来影响细胞外基质，而这种作用是由纤维母细胞增殖剂所刺激的。松弛素还会增加基质金属蛋白酶（MMP）的表达。为了研究松弛素对瘢痕疙瘩成纤维细胞和人真皮成纤维细胞中胶原蛋白和 MMP 表达的影响，作者将表达松弛素的腺病毒（dE1-RGD/lacZ/RLX）引入人真皮成纤维细胞系和瘢痕疙瘩成纤维细胞中。
方法
用 dE1-RGD/lacZ/RLX 或对照病毒感染两种成纤维细胞，通过酶联免疫吸附试验评估松弛素和分泌的转化生长因子（TGF）-β1 的蛋白水平，通过实时逆转录-聚合酶链反应和酶联免疫吸附试验评估 I 型和 III 型胶原蛋白、MMP-1 和 MMP-3 的 mRNA 水平。还检测了 Smad3 和磷酸化 Smad3 的表达，并评估了松弛素对 Smad2/3 复合物定位的影响。
结果
当人真皮成纤维细胞和瘢痕疙瘩成纤维细胞被 dE1-RGD/lacZ/RLX 或 dE1-RGD/lacZ（对照）转导时，无论是否用 TGF-β（10ng/ml）处理，松弛素均显著降低 I 型和 III 型胶原蛋白的 mRNA 表达。Smad3 和磷酸化 Smad3 的表达在瘢痕疙瘩成纤维细胞中减少，并且在 dE1-RGD/lacZ/RLX 转导后 TGF-β 会导致人真皮成纤维细胞中的 Smad2/3 复合物从胞质转位到核内，表明松弛素通过阻断 TGF-β 信号来减少胶原蛋白的合成。分析显示，dE1-RGD/lacZ/RLX 转导后，人真皮成纤维细胞和瘢痕疙瘩成纤维细胞中 MMP-1 和 MMP-3 的表达显著增加。
结论
这些结果表明，表达松弛素的腺病毒的抗纤维化作用可能对瘢痕疙瘩有治疗作用。
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