Lignin/Puerarin Nanoparticle-Incorporated Hydrogel Improves Angiogenesis through Puerarin-Induced Autophagy Activation

Abstract

Yingjing Pan,1,* Tianci Lin,1,* Longquan Shao,2 Yulin Zhang,2 Qiao Han,3 Liyuan Sheng,4 Rui Guo,5 Ting Sun,1 Yanli Zhang2 1Foshan Stomatological Hospital, School of Medicine, Foshan University, Foshan, 528225, People’s Republic of China; 2Stomatological Hospital, School of Stomatology, Southern Medical University, Guangzhou, 510280, People’s Republic of China; 3Stomatology Hospital of Guangzhou Medical University, Guangzhou, 510182, People’s Republic of China; 4Shenzhen Institute, Peking University, Shenzhen, 518057, People’s Republic of China; 5Department of Biomedical Engineering, Jinan University, Guangzhou, 510632, People’s Republic of China*These authors contributed equally to this workCorrespondence: Ting Sun, Foshan Stomatological Hospital, School of Medicine, Foshan University, Foshan, 528225, People’s Republic of China, Email [email protected] Yanli Zhang, Stomatological Hospital, School of Stomatology, Southern Medical University, Guangzhou, 510280, People’s Republic of China, Email [email protected]: Puerarin is the main isoflavone extracted from Radix Puerariae lobata (Willd.) and exerts a strong protective effect on endothelial cells. This isoflavone also exerts proven angiogenic effects; however, the potential underlying mechanism has not been fully explored. Here in this work, we aimed to determine the proangiogenesis effect of a puerarin-attached lignin nanoparticle-incorporated hydrogel and explore the underlying mechanism.Materials and Methods: Puerarin-attached lignin nanoparticles were fabricated and mixed with the GelMA hydrogel. After the hydrogel was characterized, the angiogenic effect was evaluated in a mouse hind-limb ischemia model. To further explore the mechanism of angiogenesis, human endothelial cell line EA.hy926 was exposure to different concentrations of puerarin. Wound healing assays and tube formation assays were used to investigate the effects of puerarin on cell migration and angiogenesis. qPCR and Western blotting were performed to determine the changes in the levels of angiogenesis indicators, autophagy indicators and PPARβ/δ. 3-MA was used to assess the role of autophagy in the puerarin-mediated angiogenesis effect in vivo and in vitro.Results: The hydrogel significantly improved blood flow restoration in mice with hind-limb ischemia. This effect was mainly due to puerarin-mediated increases in the angiogenic capacity of endothelial cells and the promotion of autophagy activation. A potential underlying mechanism might be that puerarin-mediated activation of autophagy could induce an increase in PPARβ/δ expression.Conclusion: The puerarin-attached lignin nanoparticle-incorporated hydrogel effectively alleviated blood perfusion in mice with hind-limb ischemia. Puerarin has a prominent proangiogenic effect. The potential mechanisms might be that puerarin-mediated autophagy activation and increase in PPARβ/δ.Graphical Abstract: Keywords: puerarin, angiogenesis, autophagy, PPARβ/δ, vascular endothelial cell

Keywords

• puerarin
• angiogenesis
• autophagy
• pparβ/δ
• vascular endothelial cell