瘢痕疙瘩成纤维细胞中Warburg效应的研究

Objective To explore whether there is a Warburg effect in keloid fibroblasts (KFs), and whether there are similar phenomena in hypertrophic scar fibroblasts (HSFs) and atrophic scar fibroblasts (ASFs). Methods Keloid, hypertrophic scar, atrophic scar and normal skin specimens were all derived from the discarded tissues of patients after surgery in the Plastic Surgery Hospital of the Chinese Academy of Medical Sciences, and each specimen was from 8 patients. KFs, HSFs, ASFs and normal skin fibroblasts (NFs) were isolated and cultured, and the glucose consumption and lactic acid production of cells in each group were detected; the relative expression levels of key glycolytic enzyme mRNAs in cells of each group were detected by qPCR; glycolysis in KFs and NFs was inhibited by 2 - deoxy - D - glucose (2 - DG), and the changes in the proliferative activities of the two types of cells were compared. Results The glucose consumption and lactic acid production in the KFs group were increased by 45.5% and 38.1% respectively compared with those in the NFs group (P < 0.05). The relative expression levels of hexokinase, pyruvate kinase and lactate dehydrogenase mRNAs in the KFs group were 4.7, 2.7 and 1.8 times those in the NFs group respectively (P < 0.05). Under the action of 2 - DG, the cell proliferative activities in the KFs group and the NFs group were decreased by 37.5% and 27.0% respectively, and the inhibitory effect of the glycolysis inhibitor on the proliferative activity of KFs was significantly higher than that in the NFs group (P < 0.05). Conclusion There is a Warburg effect in KFs, but there is no similar phenomenon in HSFs and ASFs.

目的探究瘢痕疙瘩成纤维细胞(KFs)中是否存在Warburg效应,及增生性瘢痕成纤维细胞(HSFs)、萎缩性瘢痕成纤维细胞(ASFs)是否存在类似现象。方法瘢痕疙瘩、增生性瘢痕、萎缩性瘢痕及正常皮肤标本,均来源于中国医学科学院整形外科医院患者手术后废弃组织,每种标本各来源于8例患者。分离培养KFs、HSFs、ASFs及正常皮肤成纤维细胞(NFs),检测各组细胞葡萄糖消耗量与乳酸产生量;以qPCR检测各组细胞糖酵解关键酶mRNA的相对表达量;利用2-脱氧-D-葡萄糖(2-DG)抑制KFs与NFs中糖酵解,对比2种细胞增殖活性的变化。结果KFs组的葡萄糖消耗量及乳酸产生量较NFs组分别增高达45.5%、38.1% (P0.05)。KFs组己糖激酶、丙酮酸激酶、乳酸脱氢酶mRNA的相对表达量分别为NFs组的4.7、2.7、1.8倍(P0.05)。2-DG作用下,KFs组与NFs组细胞增殖活性分别降低37.5%、27.0%,糖酵解抑制剂对KFs增殖活性的抑制作用显著高于NFs组(P<0.05)。结论KFs中存在Warburg效应,而HSFs及ASFs中无类似现象。