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目的探讨脂多糖(lipopolysaccharide,LPS)对皮肤成纤维细胞胶原代谢的影响,以了解LPS在增生性瘢痕形成中的生物学作用。方法取正常皮肤行成纤维细胞培养后,分为1个对照组及6个实验组。实验组终浓度分别为0.005、0.010、0.050、0.100、0.500和1.000μg/ml大肠杆菌LPS(E.coli055:B5)培养,对照组为DMEM培养。用逆转录-聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)测定成纤维细胞Ⅰ、Ⅲ型前胶原mRNA及胶原酶mRNA的表达,并以同一个体相同代数的瘢痕组织成纤维细胞做对照。结果与对照组比较,0.005～0.500μg/ml组促进细胞胶原合成,且0.100μg/ml组作用达高峰(P<0.05),1.000μg/ml LPS抑制细胞胶原合成(P<0.05)。LPS刺激浓度在0.005μg/ml～0.100μg/ml时,促进正常皮肤成纤维细胞Ⅰ、Ⅲ型前胶原mRNA表达,抑制胶原酶mRNA表达,且呈一定的剂量依赖性;当LPS刺激浓度为0.500μg/ml,上述作用下降;而当LPS刺激浓度到达1.000μg/ml时,抑制正常皮肤成纤维细胞Ⅰ、Ⅲ型前胶原mRNA表达,促进胶原酶mRNA表达。LPS刺激浓度在0.100μg/ml时,成纤维细胞Ⅰ、Ⅲ型前胶原mRNA和胶原酶mRNA表达与同一个体增生性瘢痕组织成纤维细胞近似。结论 LPS对人皮肤成纤维细胞Ⅰ、Ⅲ型前胶原mRNA和胶原酶mRNA表达的直接调节,可能是其参与增生性瘢痕形成的重要机制。
Objective To investigate the effect of lipopolysaccharide (LPS) on collagen metabolism of dermal fibroblasts in order to understand the biological role of LPS in the formation of hypertrophic scars. Methods Normal skin fibroblasts were cultured and divided into 1 control group and 6 experimental groups. The final concentration of the experimental group were 0.005, 0.010, 0.050, 0.100, 0.500 and 1.000μg / ml E. coli LPS (E.coli055: B5) respectively. The control group was cultured in DMEM. The expression of procollagen type Ⅰ and type Ⅲ procollagen mRNA and collagenase mRNA in fibroblasts were determined by reverse transcription-polymerase chain reaction (RT-PCR), and the same algebraic scar tissue fibroblasts Make a comparison. Results Compared with the control group, the cells in the concentration of 0.005 ~ 0.500μg / ml promoted the collagen synthesis and reached the peak at the concentration of 0.100μg / ml (P <0.05), while LPS at 1.000μg / ml inhibited the collagen synthesis (P <0.05). LPS stimulated the concentration of 0.005μg / ml ~ 0.100μg / ml, promote normal skin fibroblasts Ⅰ, Ⅲ procollagen mRNA expression, inhibition of collagenase mRNA expression, and showed a dose-dependent; when the LPS stimulus concentration of 0.500 μg / ml, the above effect decreased; when LPS stimulation concentration reached 1.000μg / ml, the normal skin fibroblasts Ⅰ, Ⅲ procollagen mRNA expression and promote collagenase mRNA expression. When the concentration of LPS was 0.100μg / ml, the mRNA expression of procollagen type Ⅰ and type Ⅲ procollagen and collagenase mRNA in fibroblasts were similar to that of fibroblasts in the same individual hypertrophic scar. Conclusion The direct regulation of LPS on the mRNA expression of procollagen type Ⅰ and type Ⅲ procollagen and collagenase in human skin fibroblasts may be an important mechanism involved in the formation of hypertrophic scars.