CAJ | 학술논문

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ⅢmRNA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(10-3 mmol/L)可降低增生性瘢痕成纤维细胞产生TGF-β1 (P＜0.05)和α-SMA mRNA、胶原Ⅰ mRNA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Ⅰ mRNA表达的抑制作用(P＜0.05)；但褪黑素对该细胞胶原ⅢmRNA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.
목적 탐토퇴흑소대증생성반흔성섬유세포생물학활성적조공.방법 분리배양증생성반흔성섬유세포,채용사담서복합물/류산분진갑지(XTT/PMS)법검측세포증식활성,매련면역흡부실험(ELISA)검측세포배양상청액중β1전화생장인자(TGF-β1)함량화형광정량취합매련반응(PCR)검측세포α평활기기동단백(α-SMA)、효원Ⅰ화효원ⅢmRNA표체,평개퇴흑소화퇴흑소수체길항제(luzindole)대증생성반흔성섬유세포생물학활성적영향.결과 퇴흑소억제증생성반흔성섬유세포적증식,억제효과정농도의뢰성(P＜0.05)；고농도퇴흑소(10-3 mmol/L)가강저증생성반흔성섬유세포산생TGF-β1 (P＜0.05)화α-SMA mRNA、효원Ⅰ mRNA적표체(P＜0.05)；퇴흑소수체길항제능조단퇴흑소대세포산생TGF-β1화α-SMA、효원Ⅰ mRNA표체적억제작용(P＜0.05)；단퇴흑소대해세포효원ⅢmRNA표체무영향(P＞0.05).결론 퇴흑소가통과여수체결합조공증생성반흔성섬유세포적생물학활성.
Objective To investigate the effect and mechanism of melatonin on bioactivity of fibroblasts 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 a-SMA,collagen Ⅰ,collagen Ⅲ mRNA were determined with real-time PCR method.Results Compared with the control,melatonin at the concentration of 10-5mmol/L,10-3mmol/L,and 1 mmol/L could inhibit the proliferation 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 a-SMA mRNA and collagen 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 activity of fibroblasts from human hypertrophic scar through a receptor pathway.