增生性瘢痕组织中褪黑素受体的表达

The expression of melatonin receptor in human hypertrophic scar

目的 检测人增生性瘢痕组织和正常皮肤中褪黑素受体含量的差异,探讨其意义.方法 人增生性瘢痕组织和正常皮肤各10例,免疫组化检测组织中褪黑素受体GPR50的表达,荧光定量PCR（SYBR Green）检测组织中褪黑素受体MT1、MT2 mRNA含量,并将PCR阳性产物基因测序.结果 免疫组化显示褪黑素受体GPR50表达于细胞膜和细胞质,正常皮肤表皮基底层细胞、毛囊和汗腺均有褪黑素受体阳性表达,增生性瘢痕组织表皮、残留的毛囊和汗腺内也存在褪黑素受体表达;同时发现增生性瘢痕真皮成纤维细胞有广泛褪黑素受体阳性表达,而正常皮肤组织中则少见表达.荧光定量PCR显示增生性瘢痕组织中MT1、MT2 mRNA含量分别为（1.16584 4±0.21829）和（0.99550 ±0.14624）拷贝数/μl cDNA,正常皮肤中为（0.99081±0.26485）和（0.77083±0.15927）拷贝数/μl cDNA,增生性瘢痕组织中MT1、MT2 mRNA含量均高于正常皮肤（P〈O.05）,同时MT1 mRNA含量在两种组织中均高于MT2 mRNA（P〈0.05）;基因测序显示扩增产物与GeneBank中人MT1、MT2受体基因序列相吻合.结论 褪黑素受体在增生性瘢痕组织和正常皮肤中均有表达,而且在增生性瘢痕组织中高于正常皮肤,褪黑素受体在增生性瘢痕形成过程中可能发挥着重要作用.

Objective To investigate the expression and its significance of melatonin receptor in human hypertrophic scarring. Methods The expression of melatonin receptor GPR50 was detected with immunohistochemistiy and the melatonin receptors（ MT1 、MT2）mRNA were assessed with RT-PCR method in 10 cases of human hypertrophic scar and normal skin. The positive production was sequenced with auto sequencing instrument. Results Positive signals of melatonin receptor could be found in the cell membrane and cytoplasm. The melatonin receptor GPR50 was located in the epithelial basal cells, sweat gland cells and hair follicle in both hypertrophic scar and normal skin. The melatonin receptor GPR50 was extensively expressed in fibroblasts of hypertrophic scar, but not in fibroblasts in normal skin. RT-PCR showed that the expression of melatonin receptor（ MT1, MT2 ） mRNA in hypertrophic scar was significantly higher than that in normal skin（ P 〈0. 05）. In normal skin and hypertrophic scar group, the expression of MT1 mRNA was higher than MT2 mRNA （ P 〈 0. 05 ） . In normal skin and hypertrophic scar group, the expression of MT1 mRNA was 0.99081 ±0.26485 and 1.16584 ±0.21829 copy number/μl cDNA, respectively;the expression of MT2 mRNA was 0. 77083 ±0. 15927and 0. 99550 ±0. 14624 copy number/ μl cDNA, respectively. Sequencing results indicated that the positive product coincided with cDNA of human melatonin receptor in GeneBank. Conclusions Positive expression of melatonin receptor can be found in human hypertrophic scar and normal skin, but it is higher in scar. The over expression of melatonin receptor in hypertrophic scar may be related to the development of hypertrophic scar.