脂多糖对增生性瘢痕患者正常皮肤成纤维细胞增殖及Ⅰ、Ⅲ型前胶原mRNA和胶原酶mRNA表达的影响  

Influence of Lipopolysaccharide on the Proliferation and the mRNAs Expression of Procollagen TypeⅠ,Ⅲ and Collagenase of Normal Skin Fibroblasts of Hypertrophic Scar Patients

作　　者：李凤玉[1] 贾国洪[1] 万立[1] 闫永宏[1] 石军[1] 

机构地区：[1]解放军第二五一医院肿瘤科,河北张家口075000

出　　处：《临床医学工程》2011年第2期161-164,共4页Clinical Medicine & Engineering

基　　金：国家自然科学基金资助项目(编号:30371467)

摘　　要：目的探讨脂多糖（hpopolysaccharide，LPS）对皮肤成纤维细胞增殖和I、Ⅲ型前胶原mRNA和胶原酶mRNA表达的影响．以了解LPS在增生性瘢痕形成中的生物学作用。方法取正常皮肤按黄勇等方法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O．005、0．010、O．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。分别通过MTT比色法及细胞计数法观察各组1—9d吸光度（A）值和细胞数量的变化；并对刺激后细胞进行传代，用逆转录一聚合酶链反应（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作对照。结果与对照组比较，0．005～0．500μg/ml组A值增加，于5～9d差异有统计学意义（P〈0．05）；1．000μg／ml组A值降低，于3～9d差异有统计学意义（P〈0．05）。与对照组比较，0．005～O．500μg／ml组细胞数量显著增加，于1～6d差异有统计学意义（P〈O．05）；1．000μg／ml组细胞数量显著降低，于2～9d差异有统计学意义（P〈0．05）。与对照组比较，0．005～O．500μg／ml组促进细胞胶原合成，且O．100μg/ml组作用达高峰（P〈O．05），1．000μg／mlLPS抑制细胞胶原合成（p〈O．05）。LPS刺激浓度在0．005～0．100μg/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O．500μg／m1，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对正常人皮肤成纤维细胞增殖及I、Ⅲ型前胶原mRNA和胶原酶mRNAObjective To observe the influence of lipopolysaccharide （LPS） on the proliferation and the mRNAs expression of pro- collagen type I, Ⅲ and collagenase of normal human skin fibroblasts and its biological role in the formation of hypertrophic sacr. Methods Fibroblasts were isolated and cultured in vitro, and then exposed to different doses of LPS （0.005, 0.010, 0.050, 0.100, 0.500, 1.000μg/ml） from E.coli055 ： B5 respectively. Then the optical density （A） values of fibroblasts were determined with the colorimeteric thiazolylblue （MTT） assay every other day from day 1 to day 9 after LPS administration, and the cell amounts were counted under inverted phase contrast microscope on a daily basis from day 1 to day 9 after LPS administration. The expression ofprocollagen type I,Ⅲ and collagenase mRNAs was tested by RT-PCR. The fibroblasts from hypertrophic scar tissue obtained from the same patients in the same culture passage were used as control. Results Compared with the control group, the A value increased with the increasing concentration of LPS （0.005 - 0.5000μg/ml） and LPS of 0.100μg/ml group had the strongest effect, the difference was remarkable at day 5 to day 9 （P〈0.05）, the A value decreased when challenged with the LPS of 1.000μg/ml and the difference was remarkable at day 3 to day 9 （P〈0.05）. The cell amount increased with the administration of LPS of different concentrations （0.005 - 0.500μg/ml） and LPS of 0.100μg/ml group had the strongest effect, the difference was remarkable at day 1 to day 6 （P〈0.05）, the cell amount decreased remarkably when challenged with LPS of 1.000μg/ml and the difference was remarkable at day 2 to day 9 （P 〈0.05）. The expression of procollagen type I ,Ⅲ mRNAs of normal skin fibroblasts in- creased, while the expression of collagenase mRNAs of normal skin fibroblasts depressed when challenged with LPS of 0.005 - 0.100μg/ml,and the influence showed a concentration dependent manner; However, when the concentration of

关 键 词：脂多糖 细胞增殖 信使核糖核酸 成纤维细胞 增生性瘢痕 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]