Liu Lin, Jiang Xiaowen, Yu Wenhui
College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China.
Key Laboratory of the Provincial Education Department of Heilongjiang for Common Animal Disease Prevention and Treatment, Harbin, China.
Evid Based Complement Alternat Med. 2019 Aug 25;2019:6027186. doi: 10.1155/2019/6027186. eCollection 2019.
In recent years, an increasing number of natural plant extracts have been determined to be potential drugs for various illnesses. In this study, we investigated the effects of dracorhodin perchlorate (DP) on fibroblast proliferation, which is crucial for wound healing. Cell proliferation assays were performed by different concentrations of DP, and the cell viability was detected by CCK-8 kits. After DP treatment for 24 h, the cell cycle was checked by flow cytometer. EGFR and downstream signaling pathways ERK1/2 and PI3K were examined with DP treatment by western blot. We further determined the effects of the related inhibitors on DP-induced relative protein phosphorylation and cell proliferation. The results showed that 3 g/mL of DP promoted cell proliferation most significantly at treatment lengths of 24 h, and the percentage of cells in the S + G2 phase increased compared to those of the control group. In western blot detection, we found that DP significantly upregulated EGFR phosphorylation and activated the downstream ERK/CREB and PI3K/Akt/mTOR signaling pathway. Moreover, the results also showed that AG1478 abolished DP-induced relative protein activation and cell proliferation. When U0126 or LY294002 pretreated cells alone, DP-induced p-ERK or p-PI3K downstream proteins and cell proliferation were suppressed compared to those of the control group, but EGFR was not affected. In addition, ICG001 and BEZ235 collectively eliminated DP-induced fibroblast proliferation. Our findings suggest that DP-promoted fibroblast proliferation is stimulated by p-EGFR-induced activation of the ERK1/2-CREB and PI3K/Akt/mTOR pathways. Our present study explored the mechanism of DP-promoted fibroblast proliferation and provided a new basis for wound healing.
近年来，越来越多的天然植物提取物被确定为治疗各种疾病的潜在药物。在本研究中，我们研究了高氯酸血竭素（DP）对成纤维细胞增殖的影响，而成纤维细胞增殖对伤口愈合至关重要。通过不同浓度的DP进行细胞增殖测定，并使用CCK-8试剂盒检测细胞活力。DP处理24小时后，通过流式细胞仪检查细胞周期。通过蛋白质印迹法检测DP处理后EGFR及其下游信号通路ERK1/2和PI3K。我们进一步确定了相关抑制剂对DP诱导的相对蛋白磷酸化和细胞增殖的影响。结果表明，在24小时的处理时间内，3μg/mL的DP最显著地促进了细胞增殖，与对照组相比，S+G2期细胞的百分比增加。在蛋白质印迹检测中，我们发现DP显著上调EGFR磷酸化并激活下游ERK/CREB和PI3K/Akt/mTOR信号通路。此外，结果还表明AG1478消除了DP诱导的相对蛋白激活和细胞增殖。当单独用U0126或LY294002预处理细胞时，与对照组相比，DP诱导的下游p-ERK或p-PI3K蛋白和细胞增殖受到抑制，但EGFR不受影响。此外，ICG001和BEZ235共同消除了DP诱导的成纤维细胞增殖。我们的研究结果表明，p-EGFR诱导的ERK1/2-CREB和PI3K/Akt/mTOR通路激活刺激了DP促进的成纤维细胞增殖。我们目前的研究探索了DP促进成纤维细胞增殖的机制，并为伤口愈合提供了新的依据。
