Liu Yueming, Liu Yang, Zeng Changchun, Li Weishan, Ke Changneng, Xu Shi
Department of Burn and Plastic Surgery, Shenzhen Longhua District Central Hospital, Affiliated Central Hospital of Shenzhen Longhua District, Guangdong Medical University, 518110 Shenzhen, Guangdong, China.
Department of Medical Laboratory, Shenzhen Longhua District Central Hospital, Affiliated Central Hospital of Shenzhen Longhua District, Guangdong Medical University, 518110 Shenzhen, Guangdong, China.
Front Biosci (Landmark Ed). 2022 Dec 7;27(12):319. doi: 10.31083/j.fbl2712319.
The aim of this study was to explore the effect of concentrated growth factor (CGF) on the wound healing potential of human epidermal cells (HaCaT) and .
CGF was extracted from venous blood using the centrifugal separation method. The CGF-conditioned medium was prepared from CGF gel immersed in Dulbecco's Modified Eagle medium. Crystal violet staining and wound healing assay were used to evaluate the proliferation and migration of HaCaT cells, respectively. Lipopolysaccharide (LPS) was used to test the anti-inflammatory function of CGF. An ELISA kit was employed to detect the concentration of growth factors and interleukins in CGF medium. mRNA and protein levels of angiogenic biomarkers (Angiopoietin-1 (ANGPT-1), vascular endothelial growth factor-A (VEGF-A) and Angiopoietin-2 (ANGPT-2) ) were determined by quantitative polymerase chain reaction (qPCR) and Western blot, respectively. A dorsal excisional wound model was recruited to test the wound healing effect of CGF in mice.
Three-day treatment of HaCaT cells with CGF significantly promoted cell proliferation, which was followed by an increase in Vascular Endothelial Growth Factor (VEGF) and Fibroblast Growth Factor (FGF) levels in the medium. Cytokines (IL-6, IL-8 and TNF-α) were increased in LPS-stimulated HaCaT cells after 3 days, and CGF slightly inhibited the mRNA expression of these cytokines. The RAS signaling pathway was activated upon CGF treatment. Both RAS knockdown and an inhibitor of RAS (zoledronic acid) could block the migration of HaCaT cells after CGF treatment. Protein expressions of CD31, ANGPT-1, and VEGF-A were up-regulated in a dose-dependent manner upon CGF exposure. The protein level of ANGPT-2 was down-regulated after CGF treatment. CGF could promote wound healing , as demonstrated using the full skin defect model in nude mice.
CGF was shown to promote wound repair and . The RAS cell signaling pathway was responsible for CGF stimulating the wound healing potential of HaCaT cells.
本研究旨在探讨浓缩生长因子（CGF）对人表皮细胞（HaCaT）伤口愈合潜能的影响。
采用离心分离法从静脉血中提取CGF。将CGF凝胶浸入杜氏改良 Eagle 培养基中制备CGF条件培养基。分别采用结晶紫染色和伤口愈合试验评估HaCaT细胞的增殖和迁移。使用脂多糖（LPS）检测CGF的抗炎功能。采用酶联免疫吸附测定（ELISA）试剂盒检测CGF培养基中生长因子和白细胞介素的浓度。分别通过定量聚合酶链反应（qPCR）和蛋白质免疫印迹法测定血管生成生物标志物（血管生成素-1（ANGPT-1）、血管内皮生长因子-A（VEGF-A）和血管生成素-2（ANGPT-2））的mRNA和蛋白质水平。采用背部切除伤口模型检测CGF对小鼠伤口愈合的作用。
用CGF处理HaCaT细胞3天可显著促进细胞增殖，随后培养基中血管内皮生长因子（VEGF）和成纤维细胞生长因子（FGF）水平升高。LPS刺激的HaCaT细胞在3天后细胞因子（IL-6、IL-8和TNF-α）增加，而CGF可轻微抑制这些细胞因子的mRNA表达。CGF处理后RAS信号通路被激活。RAS基因敲低和RAS抑制剂（唑来膦酸）均可阻断CGF处理后HaCaT细胞的迁移。CGF处理后，CD31、ANGPT-1和VEGF-A的蛋白表达呈剂量依赖性上调。CGF处理后ANGPT-2的蛋白水平下调。如在裸鼠全层皮肤缺损模型中所示，CGF可促进伤口愈合。
CGF可促进伤口修复。RAS细胞信号通路介导CGF刺激HaCaT细胞的伤口愈合潜能。
