Senior Department of Burns and Plastic Surgery, The Fourth Medical Center of PLA General Hospital, Beijing, China.
Chinese PLA Medical School, Beijing, China.
Int Wound J. 2023 May;20(5):1667-1677. doi: 10.1111/iwj.14028. Epub 2022 Dec 21.
The study aimed to explore the role of cellular communication network factor 1 (CCN1) an extracellular matrix protein in hADSC-treated wound healing. Immunofluorescence and enzyme-linked immunosorbent assays (ELISA) were used to demonstrate the secretion of CCN1 by hADSCs, isolated from human fat tissue. We investigated the role of CCN1 in wound healing by knockdown of CCN1 expression in hADSCs using CCN1 siRNA. Conditioned medium of hADSCs or hADSCs with CCN1 knocked down (hADSC-CM ) was collected. After treatment with plain DMEM/F12, hADSC-CM, hADSC-CM , or recombinant human CCN1 (rhCCN1), the wound healing abilities of human umbilical vascular endothelial cells (HUVECs) were assayed, and the AKT, also known as protein kinase B (PKB), signalling pathway was detected using western blotting. Next, we created full-thickness skin wounds on the backs of the mice and different treatments were applied to the wound surface. Wound size was measured using a digital camera on days 0-10, and evaluated. H&E and immunohistochemical staining were performed, and laser Doppler perfusion imaging was used to evaluate blood perfusion. The wound model and wound-healing assay showed that the hADSCs-CM and rhCCN1 groups had enhanced wound healing compared to the hADSCs-CM group. Further, CCN1 and hADSCs-CM promoted the proliferation and migration of HUVECs through the AKT signalling pathway. We concluded that CCN1 secreted by hADSCs enhances wound healing and promotes angiogenesis by activating the AKT signalling pathway. CCN1 plays a vital role in the regulation of hADSCs-CM during wound healing.
本研究旨在探讨细胞通讯网络因子 1（CCN1）这种细胞外基质蛋白在人脂肪来源干细胞（hADSCs）治疗创面愈合中的作用。通过酶联免疫吸附试验（ELISA）和免疫荧光检测，从人脂肪组织中分离出 hADSCs 并证实其可分泌 CCN1。通过 CCN1 siRNA 敲低 hADSCs 中的 CCN1 表达，研究 CCN1 在伤口愈合中的作用。收集 hADSCs 或敲低 CCN1 的 hADSCs（hADSC-CM）的条件培养基。在使用普通 DMEM/F12 处理后，检测 hADSC-CM、hADSC-CM 或重组人 CCN1（rhCCN1）对人脐静脉内皮细胞（HUVEC）的伤口愈合能力的影响，并通过 Western blot 检测 AKT（也称为蛋白激酶 B（PKB））信号通路。接下来，我们在小鼠背部制作全层皮肤伤口，并对伤口表面进行不同处理。在第 0-10 天使用数码相机测量伤口大小，并进行评估。进行 H&E 和免疫组织化学染色，并使用激光多普勒灌注成像评估血液灌注。伤口模型和伤口愈合试验表明，与 hADSCs-CM 组相比，hADSCs-CM 和 rhCCN1 组的伤口愈合能力增强。此外，CCN1 和 hADSCs-CM 通过 AKT 信号通路促进 HUVEC 的增殖和迁移。我们得出结论，hADSCs 分泌的 CCN1 通过激活 AKT 信号通路增强伤口愈合并促进血管生成。CCN1 在 hADSCs-CM 调节伤口愈合过程中发挥重要作用。
