目的 探讨脂多糖(lipopolysaccharide,LPS)对人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.实验组分别与终浓度为0.005、0.010、0.050、0.100、0.500和1.000 μg/ml大肠杆菌LPS(E.coli 055∶B5)培养,对照组DMEM培养.用逆转录-聚合酶链反应(RT-PCR)法测定成纤维细胞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,LPS刺激浓度在0.005～0.1μg/ml时,促进正常皮肤成纤维细胞Ⅰ、Ⅲ型前胶原mRNA表达(0.323±0.041,0.303±0.063,0.391 ±0.071,0.344±0.086,0.488±0.059,0.401 ±0.087,0.616±0.107,0.434±0.084,0.823±0.092,0.542±0.082),抑制胶原酶mRNA表达(0.598 ±0.068,0.556 ±0.049,0.441 ±0.043,0.372±0.083,0.260±0.027),且呈一定的剂量依赖性；当LPS刺激浓度为0.5μg/ml,上述作用下降(0.451±0.063,0.374±0.072,0.360±0.062)；而当LPS刺激浓度到达1.0 μg/ml时,抑制正常皮肤成纤维细胞Ⅰ、Ⅲ型前胶原mRNA表达(0.162 ±0.025,0.171 ±0.061),促进胶原酶mRNA表达(0.444±0.114).LPS刺激浓度在0.1 μg/ml时,成纤维细胞(0.823±0.092,0.542±0.082,0.260±0.027)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(0.829±0.049,0.569±0.038,0.277 ±0.059)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的表达,其直接调节可能是参与增生性瘢痕形成的重要机制.
目的 探討脂多糖(lipopolysaccharide,LPS)對人皮膚成纖維細胞膠原代謝的影響,以瞭解LPS在增生性瘢痕形成中的生物學作用.方法 取正常皮膚行成纖維細胞培養後,分為1箇對照組及6箇實驗組.實驗組分彆與終濃度為0.005、0.010、0.050、0.100、0.500和1.000 μg/ml大腸桿菌LPS(E.coli 055∶B5)培養,對照組DMEM培養.用逆轉錄-聚閤酶鏈反應(RT-PCR)法測定成纖維細胞Ⅰ、Ⅲ型前膠原mRNA及膠原酶mRNA的錶達,併以同一箇體相同代數的瘢痕組織成纖維細胞做對照.結果 與對照組比較,LPS刺激濃度在0.005～0.1μg/ml時,促進正常皮膚成纖維細胞Ⅰ、Ⅲ型前膠原mRNA錶達(0.323±0.041,0.303±0.063,0.391 ±0.071,0.344±0.086,0.488±0.059,0.401 ±0.087,0.616±0.107,0.434±0.084,0.823±0.092,0.542±0.082),抑製膠原酶mRNA錶達(0.598 ±0.068,0.556 ±0.049,0.441 ±0.043,0.372±0.083,0.260±0.027),且呈一定的劑量依賴性；噹LPS刺激濃度為0.5μg/ml,上述作用下降(0.451±0.063,0.374±0.072,0.360±0.062)；而噹LPS刺激濃度到達1.0 μg/ml時,抑製正常皮膚成纖維細胞Ⅰ、Ⅲ型前膠原mRNA錶達(0.162 ±0.025,0.171 ±0.061),促進膠原酶mRNA錶達(0.444±0.114).LPS刺激濃度在0.1 μg/ml時,成纖維細胞(0.823±0.092,0.542±0.082,0.260±0.027)Ⅰ、Ⅲ型前膠原mRNA和膠原酶mRNA錶達與同一箇體增生性瘢痕組織成纖維細胞(0.829±0.049,0.569±0.038,0.277 ±0.059)近似.結論 LPS對人皮膚成纖維細胞Ⅰ、Ⅲ型前膠原mRNA和膠原酶mRNA的錶達,其直接調節可能是參與增生性瘢痕形成的重要機製.
목적 탐토지다당(lipopolysaccharide,LPS)대인피부성섬유세포효원대사적영향,이료해LPS재증생성반흔형성중적생물학작용.방법 취정상피부행성섬유세포배양후,분위1개대조조급6개실험조.실험조분별여종농도위0.005、0.010、0.050、0.100、0.500화1.000 μg/ml대장간균LPS(E.coli 055∶B5)배양,대조조DMEM배양.용역전록-취합매련반응(RT-PCR)법측정성섬유세포Ⅰ、Ⅲ형전효원mRNA급효원매mRNA적표체,병이동일개체상동대수적반흔조직성섬유세포주대조.결과 여대조조비교,LPS자격농도재0.005～0.1μg/ml시,촉진정상피부성섬유세포Ⅰ、Ⅲ형전효원mRNA표체(0.323±0.041,0.303±0.063,0.391 ±0.071,0.344±0.086,0.488±0.059,0.401 ±0.087,0.616±0.107,0.434±0.084,0.823±0.092,0.542±0.082),억제효원매mRNA표체(0.598 ±0.068,0.556 ±0.049,0.441 ±0.043,0.372±0.083,0.260±0.027),차정일정적제량의뢰성；당LPS자격농도위0.5μg/ml,상술작용하강(0.451±0.063,0.374±0.072,0.360±0.062)；이당LPS자격농도도체1.0 μg/ml시,억제정상피부성섬유세포Ⅰ、Ⅲ형전효원mRNA표체(0.162 ±0.025,0.171 ±0.061),촉진효원매mRNA표체(0.444±0.114).LPS자격농도재0.1 μg/ml시,성섬유세포(0.823±0.092,0.542±0.082,0.260±0.027)Ⅰ、Ⅲ형전효원mRNA화효원매mRNA표체여동일개체증생성반흔조직성섬유세포(0.829±0.049,0.569±0.038,0.277 ±0.059)근사.결론 LPS대인피부성섬유세포Ⅰ、Ⅲ형전효원mRNA화효원매mRNA적표체,기직접조절가능시삼여증생성반흔형성적중요궤제.
Objective To observe the influence of lipopolysaccharide (LPS) on collagen metabolism of normal human skin fibroblasts and its biological role in the formation of hypertrophic scar.Methods Fibroblasts were isolated and cultured in vitro,and then exposed to different doses of LPS (0.005,0.01,0.05,0.1,0.5,1.0 μg/ml) from E.coli.055:B5 respectively.The expression of proccllagen type Ⅰ,Ⅲand collagenase mRNAs was tested by RT -PCR.Fibroblasts from hypertrophic scar tissue obtained from the same patients in the same culture passage were used as control.Results Compared with control group,the expression of procollagen type Ⅰ,Ⅲ mRNAs in normal skin fibroblasts increased (0.323 ± 0.041,0.303 ± 0.063,0.391 ± 0.071,0.344 ± 0.086,0.488 ± 0.059,0.401 ± 0.087,0.616 ± 0.107,0.434 ±0.084,0.823 ±0.092,0.542 ± 0.082),while the expression of collagenase mRNAs of normal skin fibroblasts depressed(0.598 ± 0.068,0.556 ± 0.049,0.441 ± 0.043,0.372 ± 0.083,0.260 ± 0.027 ).When LPS was set to the concentration of 0.005 μg/ml,it showed a concentration dependent manner.However,when the concentration of LPS was set to 0.5 μg/ml,the expression of procollagen type Ⅰ,Ⅲ and collagenase mRNAs of normal skin fibroblasts began to decrease (0.451 ± 0.063,0.374 ± 0.072,0.360 ± 0.062).When the concentration of LPS was set to 1.0 μg/ml,the expression of procollagen type Ⅰ,Ⅲ mRNAs (0.162 ± 0.025,0.171 ± 0.061 )were inhibited and the expression of collagenase mRNAs began to increase (0.444 ±0.114).When the concentration of LPS was set to 0.1 μg/ml,the expression of procollagen type Ⅰ,Ⅲ and collagenase mRNAs of normal skin fibroblasts(0.823 ±0.092,0.542 ±0.082,0.260 ±0.027)was similar to that of hypertrophic scar tissue fibroblasts(0.829 ±0.049,0.569 ±0.038,0.277 ±0.059).Conclusions This result supported that LPS may be an important factor in collagen metabolism of normal skin fibroblasts and it plays an important role in hypertrophic scar formation.
