目的:研究角质形成细胞分泌的IL-1α对成纤维细胞生物学行为的影响。方法:组织块法培养成纤维细胞,消化法培养角质形成细胞,采用免疫组化方法检测角质形成细胞分泌的IL-1α;成纤维细胞中加入含不同浓度IL-1α抗体(0.04μg/ml,0.2μg/ml,1μg/ml)的角质形成细胞条件培养液为实验组,含DMEM的条件培养液为对照组,采用Cell counting kit-8、放免法测定成纤维细胞增殖、胶原合成。结果:细胞爬片可见大量染色阳性角质形成细胞,细胞增殖测定,各实验组吸光度(A)值与对照组比较,差异均有统计学意义(P(0.01);随抗体浓度增高,A值减小,0.2μg/ml及1μg/ml浓度组与0.04μg/ml浓度组比较,差异有统计学意义(P〈0.01);胶原分泌浓度测定,各实验组与对照组比较,差异均有统计学意义(P〈0.01):随抗体浓度及胶原浓度增高,0.2μg/ml及1μg/ml浓度组与0.04μg/ml浓度组比较,差异有统计学意义(P(0.01)。结论:正常角质形成细胞分泌大量IL-1α,可促进成纤维细胞增殖,抑制胶原分泌。
Objective To observe the effects of IL-1α derived from keratinocytes on proliferation and collagen secretion of fibroblasts. Methods The immunohistochemical method was employed to confirm whether the keratinocytes can secrete IL-1α.Conditioned medium of keratinocytes with different concentration of IL-1α antibody was added to the fibroblasts,Samples were divided into the tested groups (0.04μg/ml,0.2μg/ml, 1μg/ml group)and the control (0μg/ml group). The CCK-8, radio-immunity methods were employed to measure the cell proliferation and collagen secretion. Results Keratinocytes did secret IL-1α. In fibroblast proliferation, absorbency (A value) of tested groups was different from the control's (P〈0.01). A value decreased as the concentration increasing(P〈0.01), the difference between 0.2μg/ml group, 1μg/ml group and 0.04μg/ml group had statistic meaning, but the difference between 0.2μg/ml group and 1μg/ml group had no statistic meaning. In collagen secretion, the concentration of Ⅲ procollagen increased as the concentration increasing(P〈0.01),the difference between 0.2μg/ml group, 1μg/ml group and 0.04μg/ml group had statistic meaning, but the difference between 0.2μg/ml group and 1μg/ml group had no statistic meaning. Conclusion keratinocytes can secret IL-1α which can increase fibroblasts proliferation ,inhibit collagen sercetion .