Abstract：
BACKGROUND: Migration of keratinocyte plays an essential role in wound healing. The proprietary platelet-rich plasma from human blood, named as self-growth colony (SGC), functions in stimulating migration of wounded keratinocytes. In addition, the growth factors, including VEGF, being enriched in SGC could account for this function. Scutellarin, an active phytochemical from root of Scutellaria barbata D. Don, has been proposed to have various pharmacological functions; however, the activity in epidermal skin cells is yet to be explored. Here, the role of scutellarin in potentiating the functionality of SGC to promote the regeneration of wounded keratinocyte was probed.
METHODS: Molecular docking and ultrafiltration-based LC-MS were performed to verify the binding between scutellarin and VEGF, which potentiated the VEGF-mediated functions. Scratch assay, performed on cultured keratinocytes, was to analyze the treatments of SGC and scutellarin in the process of wound healing. Western blot analysis was to confirm the involvement of signaling cascades in observed effects.
RESULTS: We have identified the binding of scutellarin with VEGF. The binding accounted for the potentiation role of scutellarin in skin regeneration, as triggered by SGC. The co-treatment of scutellarin and SGC onto scratched keratinocyte cultures was able to enhance the process of wound healing, that is, scutellarin showed a potentiating effect to SGC. In addition, the potentiation of scutellarin was shown to be mediated by phosphorylation of VEGF receptor-2 (VEGFR2) and mitogen-activated protein kinase (MAPK) signaling.
CONCLUSIONS: These findings support the application of scutellarin as an enhancing agent in potentiating the SGC-mediated wound healing.
METHODS: Molecular docking and ultrafiltration-based LC-MS were performed to verify the binding between scutellarin and VEGF, which potentiated the VEGF-mediated functions. Scratch assay, performed on cultured keratinocytes, was to analyze the treatments of SGC and scutellarin in the process of wound healing. Western blot analysis was to confirm the involvement of signaling cascades in observed effects.
RESULTS: We have identified the binding of scutellarin with VEGF. The binding accounted for the potentiation role of scutellarin in skin regeneration, as triggered by SGC. The co-treatment of scutellarin and SGC onto scratched keratinocyte cultures was able to enhance the process of wound healing, that is, scutellarin showed a potentiating effect to SGC. In addition, the potentiation of scutellarin was shown to be mediated by phosphorylation of VEGF receptor-2 (VEGFR2) and mitogen-activated protein kinase (MAPK) signaling.
CONCLUSIONS: These findings support the application of scutellarin as an enhancing agent in potentiating the SGC-mediated wound healing.
摘要：
背景：角质形成细胞的迁移在伤口愈合中起着至关重要的作用。来自人类血液的富含血小板的专有血浆，被命名为自我生长菌落(SGC)，在刺激受伤的角质形成细胞迁移中的功能。此外,增长因素，包括VEGF,丰富SGC可以解释这一功能。Scutellarin,一种来自半枝莲D.Don根的活性植物化学物质，已提出具有各种药理功能；然而，表皮皮肤细胞的活性还有待探索。这里,探讨了灯盏乙素在增强SGC功能以促进受伤角质形成细胞再生中的作用。
方法：进行分子对接和基于超滤的LC-MS以验证灯盏乙素与VEGF之间的结合，增强了VEGF介导的功能。划痕试验,在培养的角质形成细胞上进行，分析SGC和灯盏乙素在创面愈合过程中的治疗效果。Western印迹分析是为了证实所观察到的效应中信号级联的参与。
结果：我们已经确定了灯盏乙素与VEGF的结合。该结合解释了灯盏乙素在皮肤再生中的增强作用，由SGC触发。将灯盏乙素和SGC共同处理在刮伤的角质形成细胞培养物上能够增强伤口愈合的过程，也就是说,灯盏乙素对SGC有增强作用。此外,显示灯盏乙素的增强作用是由VEGF受体2(VEGFR2)和丝裂原活化蛋白激酶(MAPK)信号的磷酸化介导的.
结论：这些发现支持灯盏乙素作为增强剂在增强SGC介导的伤口愈合中的应用。
方法：进行分子对接和基于超滤的LC-MS以验证灯盏乙素与VEGF之间的结合，增强了VEGF介导的功能。划痕试验,在培养的角质形成细胞上进行，分析SGC和灯盏乙素在创面愈合过程中的治疗效果。Western印迹分析是为了证实所观察到的效应中信号级联的参与。
结果：我们已经确定了灯盏乙素与VEGF的结合。该结合解释了灯盏乙素在皮肤再生中的增强作用，由SGC触发。将灯盏乙素和SGC共同处理在刮伤的角质形成细胞培养物上能够增强伤口愈合的过程，也就是说,灯盏乙素对SGC有增强作用。此外,显示灯盏乙素的增强作用是由VEGF受体2(VEGFR2)和丝裂原活化蛋白激酶(MAPK)信号的磷酸化介导的.
结论：这些发现支持灯盏乙素作为增强剂在增强SGC介导的伤口愈合中的应用。
