真皮与脂肪组织来源成纤维细胞致纤维化能力的比较研究

Fibrogenesis between dermal and adipose tissue-derived fibroblasts

目的初步探讨真皮与脂肪组织来源的成纤维细胞的致纤维化能力及其作用机制。方法自同一杜洛克雌性猪真皮和皮下脂肪组织中分别分离和培养成纤维细胞,利用倒置显微镜和透射电子显微镜观察细胞形态和超微结构,采用Real-TimePCR技术检测细胞Ⅰ型和Ⅲ型前胶原、α平滑肌肌动蛋白(α-SMA)、转化生长因子β1(TGF-β1)和类胰岛素样生长因子1(IGF-1)mRNA的相对表达量。结果与脂肪来源的成纤维细胞比较,真皮组织来源的成纤维细胞胞体较大,粗面内质网扩张明显。Real-Time PCR检测结果显示:真皮组织来源的成纤维细胞中Ⅰ型前胶原和α-SMA的mRNA相对表达量均显著高于脂肪来源的成纤维细胞(P<0.05),而Ⅲ型前胶原和IGF-1的mRNA相对表达量显著低于脂肪来源的成纤维细胞(P<0.05);两种不同组织来源成纤维细胞中TGF-β1 mRNA相对表达量比较,差异无统计学意义(P>0.05)。结论真皮与脂肪组织来源的成纤维细胞在与致纤维化相关的细胞形态和功能方面均存在差异。在真皮和脂肪组织受到创伤后的纤维化进程中,两种组织来源的成纤维细胞致纤维化的起点已存在差异。

Objective To investigate the fibrogenesis of dermal and adipose tissue-derived fibroblasts,and explore the mechanism. MethodsFibroblasts derived from dermal tissues and subcutaneous adipose tissues in the same pig were isolated and cultured.Inverted microscope and transmission electron microscope were used to observe the cell morphology and ultrastructure,and Real-Time PCR was employed to detect the relative expression of procollagen type ⅠmRNA,procollagen type Ⅲ mRNA,α-smooth muscle actin（α-SMA） mRNA,transforming growth factor β1（TGF-β1） mRNA and insulin-like growth factor-1（IGF-1） mRNA in cells. ResultsCompared with adipose tissue-derived fibroblasts,the cell body of dermal tissue-derived fibroblasts was larger,with expanded rough endoplasmic reticulum.The relative expression of precollagen type I mRNA and α-SMA mRNA in dermal tissue-derived fibroblasts was significantly higher than that in adipose tissue-derived fibroblasts（P0.05）,while the relative expression of precollagen type Ⅲ mRNA and IGF-1 mRNA in dermal tissue-derived fibroblasts was significantly lower than that in adipose tissue-derived fibroblasts（P0.05）.There was no significant difference in the relative expression of TGF-β1 mRNA between adipose tissue-derived fibroblasts and dermal tissue-derived fibroblasts（P0.05）. ConclusionThere are differences in cell morphology and function concerning with fibrogenesis between dermal tissue-derived fibroblasts and adipose tissue-derived fibroblasts.The starting points of wound healing process after dermal and adipose tissue impairment are different between dermal tissue-derived fibroblasts and adipose tissue-derived fibroblasts.