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

摘要

Objective To explore the effect of activin receptor-like kinase( ALK )5 inhibition on the expression of collagen type Ⅰ( COL1A2 )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 COL1A2 and a-SMA in the fibroblasts were determined by quantitative reverse transcription PCR and Western blot respectively. Results Compared with control group,COLIA2 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 a-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 a-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.%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、10 μM)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.