大成散对成纤维细胞增殖活性及鞘氨醇激酶/1-磷酸鞘氨醇通路的影响

目的 研究大成散对体外培养人皮肤成纤维细胞（HFF-1）增殖及鞘氨醇激酶/1-磷酸鞘氨醇（SPHK/S1 P）通路的影响，并初步探讨其作用机制。方法 制备大成散药液，采用CCK-8法检测不同浓度大成散药液对HFF-1细胞增殖活性的影响，确定最佳干预时间与浓度进行后续实验，Western Blot法检测大成散药液对HFF-1细胞转化生子因子-β1（TGF-β1）、SPHK1、SPHK2、S1 P蛋白表达的影响，qRT-PCR法检测大成散药液对HFF-1细胞TGF-β1、SPHK1、SPHK2 mRNA表达的影响。结果 5×10-2 g/L的大成散干预48 h可促进HFF-1细胞的增殖，提高细胞活力；与对照组比较，大成散可以增加HFF-1细胞中TGF-β1、SPHK1、SPHK2、S1 P蛋白表达水平（P<0.05），提高HFF-1细胞中TGF-β1、SPHK1、SPHK2 mRNA表达水平（P<0.05）。结论 大成散可通过促进成纤维细胞增殖，上调TGF-β1表达，激活SPHK1/S1 P通路，促进血管新生，发挥抗炎效应，或通过激活SPHK2，催化产生S1 P，启动缺氧预处理机制，从而保护皮肤成纤维细胞，提高成纤维细胞的增殖能力，进而加速创面愈合。

Effect of Dacheng Powder on Proliferative Activity of HFF-1 and Sphingosine Kinase/Sphingosine 1-phosphate Pathway

Objective To study the effect of Dacheng Powder on the proliferation of human foreskin fibroblasts-1 （HFF-1） and sphingosine kinase/Sphingosine 1-phosphate（SPHK/S1 P）pathway of human skin fibroblasts cultured in vitro， and to preliminarily study its mechanism of action. Methods Dacheng Powder liquid was prepared. The effects of different concentrations of Dacheng Powder liquid on the proliferation activity of HFF-1 cells were detected by Cell Counting Kit-8 （CCK-8）， and the optimal intervention conditions were determined for subsequent experiments. Western Blot was used to detect the effects of Dacheng Powder liquid on expressions of transforming growth factor beta1 （TGF-β1），sphingosine kinase （SPHK1）， SPHK2， sphingosine 1-phosphate （S1 P） protein. Quantitative Real-time polymerase chain reaction （qRT-PCR） method was used to detect Dacheng Powder on mRNA expressions of TGF-β1， SPHK1， and SPHK2. Results Dacheng Powder at 5×10-2 g/L promoted the proliferation of HFF-1 cells and improved cell viability. Compared with the control group， Dacheng Powder increased the protein expressions of TGF-β1， SPHK1， SPHK2， and S1P in HFF-1 cells（P<0.05）， and elevated mRNA expressions of TGF-β1， SPHK1， SPHK2 in HFF-1 cells（P<0.05）. Conclusion Dacheng Powder exerted anti-inflammatory effects possibly through promoting the proliferation of fibroblasts， up-regulating the expression of TGF-β1， activating the SPHK1/S1P pathway， promoting angiogenesis， or protecting skin fibroblasts and improving the proliferation of fibroblasts， thereby accelerating wound healing possibly through activating SPHK2， catalyzing the production of S1P， and initiating the mechanism of hypoxia preconditioning.