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论著:系统性硬皮病患者外周血单个核细胞对皮肤克隆成纤维细胞胶原代谢的影响
Effects of Peripheral Blood Mononuclear Cells on the Collagen Metabolism of Cloned Systemic Scleroderma Skin Fibroblasts
高地 朱鹭冰 李明
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作者单位:复旦大学附属中山医院皮肤科
中文关键字:系统性硬皮病;成纤维细胞;克隆细胞;胶原;外周血单个核细胞
英文关键字:Systemic scleroderma; Fibroblasts; Clone cells; Collagen; Peripheral blood mononuclear cell
中文摘要:目的:探讨系统性硬皮病(systemic scleroderma, SSc)患者外周血单个核细胞(peripheral blood mononuclear cells, PBMCs)对具有不同胶原合成能力的皮肤克隆成纤维细胞的胶原代谢的影响。方法:采用实时荧光定量逆转录 聚合酶链反应(reverse transcription polymerase chain reaction, RT PCR)法检测系统性硬皮病患者和健康对照者PBMC培养上清液对具有不同胶原合成能力的皮肤克隆成纤维细胞中的Ⅰ型前胶原和基质金属蛋白酶 1(matrix metalloproteinase 1,MMP 1)的mRNA水平的影响。结果:在体外培养的系统性硬皮病皮肤克隆成纤维细胞中加入10%患者PBMC培养上清液继续培养48 h,仅使具有低胶原合成能力的SSc克隆成纤维细胞的Ⅰ型前胶原和MMP 1的mRNA水平上升[(0.70±0.29)比(0.07±0.08), P=0.001;(0.75±0.16) 比(0.09±0.20),P<0.001];而加入10%健康对照PBMC培养上清液继续培养48h,仅使具有高胶原合成能力的SSc克隆成纤维细胞的Ⅰ型前胶原的mRNA水平下降[( 0.24±0.08)比(0.12±0.10), P=0.024],同时使具有高、低胶原合成能力的SSc克隆成纤维细胞的MMP 1 mRNA 水平上升(P=0.003;P<0.001),且具有低胶原合成能力的克隆成纤维细胞MMP 1 mRNA水平上升幅度大[(1.27±0.23)比(0.56±0.08),P=0.000]。结论:SSc具有不同胶原合成能力的皮肤克隆成纤维细胞的Ⅰ型前胶原和MMP 1的mRNA水平对PBMC的反应存在异质性。
英文摘要:Objective: To study the effects of peripheral blood mononuclear cells (PBMCs) on the collagen metabolism of cloned systemic scleroderma (SSc) skin fibroblasts with heterogeneity in the synthesis of collagen. Methods:By means of real time reverse transcription polymerase chain reaction (RT PCR), the effects of PBMC supernatants (from SSc patients and controls) on the expression of type Ⅰ procollagen and matrix metalloproteinase 1 (MMP 1) on mRNA level in cloned SSc skin fibroblasts with heterogeneity in the synthesis of collagen were determined. Results: Type Ⅰ procollagen and MMP 1 mRNA levels of cloned SSc fibroblasts were upregulated by PBMC from SSc patients (10%, 48 h) only in low collagen producing clones [(0.70±0.29) vs. (0.07±0.08), P=0.001; (0.75±0.16) vs. (0.09±0.20),P<0.001]. Type Ⅰ procollagen mRNA levels of cloned SSc fibroblasts were downregulated by PBMC from controls (10%, 48 h) only in high collagen producing clones [( 0.24±0.08) vs. (0.12±0.10), P=0.024]. MMP 1 mRNA levels of cloned SSc fibroblasts were upregulated by PBMC from controls(10%, 48h) in both high and low collagen producing clones (P=0.003, P<0.001), and the increase was greater in low collagen producing clones than in high collagen procuding clones [(1.27±0.23) vs. (0.56±0.08),P=0.001]. Conclusions: Type Ⅰ procollagen and MMP 1 mRNA levels of cloned SSc skin fibroblasts with heterogeneity in the synthesis of collagen are heterogeneous in their response to PBMCs.
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