褪黑素对增生性瘢痕成纤维细胞生物学活性的调控

Effect of melatonin on bioactivity of fibroblasts from human hypertrophic scar

目的探讨褪黑素对增生性瘢痕成纤维细胞生物学活性的调控。方法分离培养增生性瘢痕成纤维细胞，采用四氮唑复合物／硫酸酚嗪甲酯（XTT／PMS）法检测细胞增殖活性，酶联免疫吸附实验（ELISA）检测细胞培养上清液中β1转化生长因子（TGF-β1）含量和荧光定量聚合酶链反应（PCR）检测细胞α平滑肌肌动蛋白（α-SMA）、胶原Ⅰ和胶原Ⅲ mRNA表达，评价褪黑素和褪黑素受体拮抗剂（luzindole）对增生性瘢痕成纤维细胞生物学活性的影响。结果褪黑素抑制增生性瘢痕成纤维细胞的增殖，抑制效果呈浓度依赖性（P〈0．05）；高浓度褪黑素（10^-3mmol／L）可降低增生性瘢痕成纤维细胞产生TGF-β1（P〈0．05）和α-SMAmRNA、胶原ⅠmRNA的表达（P〈0．05）；褪黑素受体拮抗剂能阻断褪黑素对细胞产生TGF-β1和α-SMA、胶原ⅠmRNA表达的抑制作用（P〈0．05）；但褪黑素对该细胞胶原Ⅲ mRNA表达无影响（P〉0．05）。结论褪黑素可通过与受体结合调控增生性瘢痕成纤维细胞的生物学活性。

Objective To investigate the effect and mechanism of melatonin on bioactivity of fi- broblasts from human hypertrophic scar. Methods Fibroblasts from hypertrophic scar were cultured and incubated with melatonin, melatonin and Luzindole, and Luzindole, respectively, for 24 h, and the media as control. XTT/PMS assay was used to measure the proliferation of fibroblasts, ELISA assay to detect the TGF-β1 production of fibroblasts, and the expression of cell α-SMA, collagen Ⅰ , collagen Ⅱ mRNA were determined with real-time PCR method. Results Compared with the control, melatonin at the concentration of 10^-5 mmol/L, 10^-3mmol/L, and 1 mmol/L could inhibit the prolifera- tion of fibroblasts in a does-dependent manner （P〈0.05）; melatonin at the concentration of 10^-3 mmol/L could significantly decrease the TGF-β1 production and expression of α-SMA mRNA and col- lagen Ⅰ mRNA in fibroblasts from human hypertrophic scar （P〈0.05） ; the effect of melatonin on fibroblast was significantly blocked by Luzindole （P〈0.05）, but melatonin could not inhibit collagen Ⅲ mRNA expression （P〉0.05）. Conclusions Melatonin can significantly regulate the biological ac- tivity of fibroblasts from human hypertrophic scar through a receptor pathway.