背景:创伤导致的伤口不愈合,手术,慢性疾病每年影响全球数百万人,由于对控制组织修复和再生的分子过程的理解不完全,因此可用的治疗策略有限。丹酚酸B(SalB)在促进血管生成和抑制炎症方面显示出有希望的生物活性。然而,其在组织再生中的调控机制尚不清楚。
目的:本研究旨在研究SalB对伤口愈合和再生过程的影响,连同其潜在的分子机制,通过使用斑马鱼作为模型生物。
方法:在本研究中,我们采用了多方面的方法来评估SalB对斑马鱼尾鳍再生的影响。我们利用了全鱼免疫荧光,TUNEL染色,线粒体膜电位(MMP),和吖啶橙(AO)探针分析SalB治疗下的组织修复和再生。此外,我们利用转基因斑马鱼菌株研究了在鳍再生的不同阶段炎症细胞的迁移。为了验证小窝蛋白-1(Cav1)在组织再生中的重要性,我们使用分子对接和基于吗啉代的基因敲低技术探讨了其功能作用。此外,我们通过应用原位杂交定量Cav1表达水平。
结果:我们的研究结果表明,SalB通过促进细胞增殖的多方面机制加速斑马鱼尾鳍再生,抑制细胞凋亡,和增强MMP。此外,发现SalB在组织再生过程中对免疫细胞的动态聚集和随后的消退施加调节控制。重要的是,我们观察到Cav1的敲低显著损害了组织再生,导致免疫细胞过度浸润和凋亡水平增加。此外,Cav1的击倒也会影响胚层的形成,Cav1影响组织再生的关键过程。
结论:这项研究的结果表明,SalB通过调节免疫细胞迁移和Cav1介导的成纤维细胞活化来促进组织修复和再生,促进胚芽的形成和发展。这项研究强调了SalB在促进组织再生方面的潜在药理作用。这些发现促进了再生医学研究的发展和创伤新治疗方法的发展。
BACKGROUND: Non-healing wounds resulting from trauma, surgery, and chronic diseases annually affect millions of individuals globally, with limited therapeutic strategies available due to the incomplete understanding of the molecular processes governing tissue repair and regeneration. Salvianolic acid B (Sal B) has shown promising bioactivities in promoting angiogenesis and inhibiting inflammation. However, its regulatory mechanisms in tissue regeneration remain unclear.
OBJECTIVE: This study aims to investigate the effects of Sal B on wound healing and regeneration processes, along with its underlying molecular mechanisms, by employing zebrafish as a model organism.
METHODS: In this study, we employed a multifaceted approach to evaluate the impact of Sal B on zebrafish tail fin regeneration. We utilized whole-fish immunofluorescence, TUNEL staining, mitochondrial membrane potential (MMP), and Acridine Orange (AO) probes to analyze the tissue repair and regenerative under Sal B treatment. Additionally, we utilized transgenic zebrafish strains to investigate the migration of inflammatory cells during different phases of fin regeneration. To validate the importance of Caveolin-1 (Cav1) in tissue regeneration, we delved into its functional role using molecular docking and Morpholino-based gene knockdown techniques. Additionally, we quantified Cav1 expression levels through the application of in situ hybridization.
RESULTS: Our findings demonstrated that Sal B expedites zebrafish tail fin regeneration through a multifaceted mechanism involving the promotion of cell proliferation, suppression of apoptosis, and enhancement of MMP. Furthermore, Sal B was found to exert regulatory control over the dynamic aggregation and subsequent regression of immune cells during tissue regenerative processes. Importantly, we observed that the knockdown of Cav1 significantly compromised tissue regeneration, leading to an excessive infiltration of immune cells and increased levels of apoptosis. Moreover, the knockdown of Cav1 also affects blastema formation, a critical process influenced by Cav1 in tissue regeneration.
CONCLUSIONS: The results of this study showed that Sal B facilitated tissue repair and regeneration through regulating of immune cell migration and Cav1-mediated fibroblast activation, promoting blastema formation and development. This study highlighted the potential pharmacological effects of Sal B in promoting tissue regeneration. These findings contributed to the advancement of regenerative medicine research and the development of novel therapeutic approaches for trauma.