脂多糖对人正常皮肤成纤维细胞增殖周期及Ⅰ、Ⅲ型前胶原mRNA表达的影响  

Influence of lipopolysaccharide on the cell cycle and the mRNAs expression of procollagen type Ⅰ, Ⅲ of normal human skin fibroblasts

作　　者：王舒琦[1] 李凤玉[1] 万立[1] 闫淑英[1] 张建宇[1] 杨红明[2] 

机构地区：[1]解放军第二五一医院肿瘤科,河北省张家口075000 [2]解放军总医院第一附属医院全军烧伤研究所

出　　处：《中国医师杂志》2011年第7期865-868,共4页Journal of Chinese Physician

基　　金：国家自然科学基金（3071467）

摘　　要：目的观察大肠杆菌脂多糖（lipopolysaccharide，LPS）刺激后，皮肤成纤维细胞增殖周期和Ⅰ、Ⅲ型前胶原mRNA表达变化规律。方法体外培养正常人皮肤成纤维细胞，加入不同浓度（0．005～1．0μg／m1）的大肠杆菌LPS（Ecoli055：B5），于LPS加入后第7天细胞处于对数生长期时，采用流式细胞术观察细胞增殖周期；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。结果LPS刺激浓度在0．005～0．1μg／ml时，S期细胞比例随着刺激浓度增加而增高，且呈浓度依赖性；当LPS浓度为0．5μg／ml时，上述作用开始降低，但仍高于空白对照；当浓度达到1．0μg／ml时，s期细胞比例均低于空白对照；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，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0．5μg／ml，上述作用下降；而当LPS刺激浓度到达1．0μg/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。结论在一定浓度范围内LPS促进皮肤成纤维细胞增殖和胶原合成，而过高浓度LPS则呈现抑制效应。Objective To observe the influence of lipopolysaccharide （LPS） on the cell cycle and the mRNAs expression of procollagen type Ⅰ, Ⅲ of normal human skin fibroblasts. Methods Purified dermal fibroblasts were exposed to different doses of LPS（0. 005 - 1.0 μg/ml） from E. coli. Then the cell cycle of fibroblasts at logarithmic stage at day 7 after LPS administration was assayed with flow cytometry. The expression of procollagen type Ⅰ, Ⅲ and collagenase mRNAs was tested by RT-PCR. Results The percentage of S phase cells in cell cycle of normal human skin fibroblasts increased when LPS concentrations were changed from 0. 005 to 0. 1 μg/ml, arid the increase showed a concentration dependent manner. However, when the concentration of LPS was 0. 5 μg/ml, the percentage of S phase cells began to decrease, but still higher than normal control. When LPS concentration reached 1.0μg/ml, the percentage of S phase cells were lower than normal control. The expression of procollagen type Ⅰ, Ⅲ mRNAs of normal skin fibroblasts increased when LPS was challenged to the concentration of 0. 005 μg/ml, and the influence showed a concentration dependent manner. However, when the concentration of LPS was 0. 5 μg./ml, the influence of LPS on the expression of procollagen type Ⅰ, Ⅲ of normal skin fibroblasts began to decrease. When the concentration of LPS reached 1.0 μg/ml, the expression of procollagen type Ⅰ, Ⅲ mRNAs were inhibited. Conclusions LPS promoted the proliferation and collagen synthesis of normal human skin fibroblasts within a certain range of low doses, while high doses of LPS might inhibit the proliferation and collagen synthesis of normal human skin fibroblasts.

关 键 词：脂多糖类/药理学 皮肤/细胞学/药物作用/代谢 成纤维细胞/代谢/病理学/药物 作用 细胞增殖 胶原Ⅰ型/代谢 胶原Ⅲ型/代谢 

分 类 号：R285.5[医药卫生—中药学]