丹参素对成纤维细胞生物学作用机制的研究

A study on the mechanism of the biological roles of danshensu on fibroblast

目的 探讨丹参素对成纤维细胞凋亡和前胶原基因表达的影响。 方法 于培养的人皮肤成纤维细胞（2×106）中加入丹参素（0.025 mg/ml），培养8 h后，从细胞中分离核蛋白及总RNA；采用 EMSA法测定NF-κB及 NF-l核转录因子结合活性；凋亡用 DNA梯度片段法分析；RT-PCR用于分析前胶原基因表达。 结果 丹参素对培养成纤维细胞作用8 h后，NF-κB结合活性几乎被完全抑制，NF-1结合活性降低近50％，琼脂糖凝胶电泳显示出清晰的梯状 DNA片段，RT-PCR方法测定显示，I型前胶原αl和α2的mRNA水平分别降低了56%和59%。 结论 丹参素抑制成纤维细胞核转录因子NF-kB的活性并诱导其发生凋亡，抑制成纤维细胞核转录因子NF-l的活性而调控胶原的合成与分泌，这可能是其抑制增生性瘢痕的细胞分子生物学机制。

Objective To explore the effects of danshensu on fibroblast apoptosis and the expression of procollagen gene. Methods Danshensu was added to in vitro culture of human cutaneous fibroblasts,and the nuclear proteins and total RNA were harvested from the cells. Electrophoretic mobility shift assay (EMSA) was adopted to determine the combining activity of nuclear transcription factors NF-kB and NF-1. Cell apoptosis was analyzed by DNA ladder fragmentation. The procollagen gene expression was analyzed by RT-PCR. Results Eight hours after the action of danshensu on the culture fibroblasts, the NF-kB combining activity was almost inhibited completely and the NF-1 combining activity decreased by about 50%. In addition, ladder-like DNA fragments were revealed clearly by sepharose electrophoresis. The mRNA levels of type Ⅰprocollagen α1 and α2 decreased by 56% and 59%,respectively. Conclusion Danshensu could inhibit the nuclear transcription factor NF-kB activity of fibroblast and induce the occurrence of its apoptosis. Furthermore, danshensu could also inhibit the nuclear transcription factor NF-1 activity of fibroblast and modulate the synthesis and secretion of collagen.