抑制活化素受体样激酶5对增生性瘢痕成纤维细胞中胶原蛋白沉积的影响

Effect of activin receptor-like kinase 5 inhibition on the collagen deposition in the fibroblasts of hyperplastic scars

目的研究抑制活化素受体样激酶(ALK)5对增生性瘢痕成纤维细胞Ⅰ型胶原蛋白(COL1A2)和α-平滑肌肌动蛋白(α-SMA)表达的影响。方法手术取增生性瘢痕组织进行成纤维细胞体外原代培养,采用不同浓度(1、5、10μM)的ALK5抑制剂CP-639180对增生性瘢痕成纤维细胞干预3 h后,分别采用定量逆转录PCR和Western blot方法检测Ⅰ型胶原蛋白和α平滑肌肌动蛋白的表达。结果与对照组比较,ALK5抑制剂处理后,成纤维细胞中COL1A2的mRNA和蛋白含量均明显降低,且COL1A2的mRNA和蛋白水平与抑制剂的浓度呈反比(P<0.05,P<0.01)。同样,ALK5抑制剂在转录水平和蛋白翻译水平降低了瘢痕成纤维细胞中α-SMA的表达(P均<0.05)。结论应用小分子ALK5抑制剂CP-639180可以抑制增生性瘢痕成纤维细胞分泌Ⅰ型胶原蛋白和α-SMA,进一步抑制胶原纤维的合成,为增生性瘢痕治疗研究提供新的思路。

Objective To explore the effect of activin receptor-like kinase（ALK）5 inhibition on the expres- sion of collagen type I（ COLIA2 ）and a-smooth muscle actin（α-SMA） in the fibroblasts of hyperplastic scars. Methods Hyperplastic scar tissues were used for fibroblasts cultured in vitro. After treated with different concentrations （ 1,5, 10 μM）of ALK5 inhibitor CP-639180 for 3 h, the mRNA and protein levels of COL1 A2 and α-SMA in the fibroblasts were determined by quantitative reverse transcription PCR and Western blot respectively. Results Compared with con- trol group, COL1 A2 mRNA and protein levels in the fibroblasts treated with ALK5 inhibitor were significantly reduced, and COL1A2 expression was inversely proportional to the inhibitor concentration （P 〈 0. 05, P 〈 0.01 ）. Similarly, ALK5 inhibitors reduced α-SMA expression at both transcriptional level and translation level in the fibroblasts （P 〈 0.05 ）. Conclusion Application of a small molecular inhibitor of ALK 5 CP-639180 appears to significantly inhibit collagen and α-SMA expression in the fibroblasts of hyperplastic scars. It further inhibits the synthesis of collagen fibers and may provide a novel approach to reducing proliferative scars in human.