Shen Chengcheng, Lu Yuangang, Zhang Jianghe, Li Yujie, Zhang Yiming, Fan Dongli
Department of Plastic and Cosmetic Surgery, Xinqiao Hospital, Army Medical University, Chongqing, China.
Department of Plastic and Cosmetic Surgery, Daping Hospital, Army Medical University, Chongqing, China.
Cell Transplant. 2021 Jan-Dec;30:963689721989605. doi: 10.1177/0963689721989605.
The chronic wound induced by diabetes has poor efficacy and could lead to amputation. The repair function of mesenchymal stem cells (MSCs) impaired after long-term culture . Studies have shown that the proto-oncogene c-Casitas b-lineage lymphoma (c-Cbl) can regulate receptor- and non-receptor tyrosine kinase, which was also involved in the angiogenesis process. This study aimed to explore the regulative effect of c-Cbl on the proangiogenic functions of long-term cultured MSCs and evaluate its pro-healing effect on diabetic wounds. In this study, the c-Cbl level was downregulated by locked nucleic acid-modified antisense oligonucleotide gapmers (LNA Gapmers). We detected the effect of c-Cbl downregulation on long-term cultured MSCs in terms of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signal, cellular proliferation, senescence, migration, and angiogenic factors paracrine activity . , we observed the pro-healing effect of long-term cultured MSCs, with or without c-Cbl downregulation, on the diabetic wound. We found that the phosphorylation level of c-Cbl increased and that of Akt decreased in passage 10 (P10) MSCs compared with passage 3 (P3) MSCs ( < 0.05). Additionally, the proliferation, paracrine, and migration capacity of P10 MSCs decreased significantly, accompanied by the increase of cellular senescence ( < 0.05). However, these functions, including PI3K/Akt activity of P10 MSCs, have been improved by c-Cbl downregulation ( < 0.05). Compared with P10 MSCs treatment, treatment with c-Cbl downregulated P10 MSCs accelerated diabetic wound healing, as defined by a more rapid wound closure ( < 0.05), more neovascularization ( < 0.05), and higher scores of wound histological assessment ( < 0.05) in a diabetic rat model. Our findings suggested that c-Cbl downregulation could attenuate the impairment of proangiogenic functions in MSCs induced by long-term culture and improve the effect of long-term cultured MSCs in promoting diabetic wound healing.
糖尿病引发的慢性伤口疗效不佳，可能导致截肢。间充质干细胞（MSCs）的修复功能在长期培养后受损。研究表明，原癌基因c-Casitas b系淋巴瘤（c-Cbl）可调节受体型和非受体型酪氨酸激酶，其也参与血管生成过程。本研究旨在探讨c-Cbl对长期培养的MSCs促血管生成功能的调节作用，并评估其对糖尿病伤口的促愈合作用。在本研究中，通过锁核酸修饰的反义寡核苷酸间隙mers（LNA Gapmers）下调c-Cbl水平。我们从磷脂酰肌醇3激酶（PI3K）/蛋白激酶B（Akt）信号、细胞增殖、衰老、迁移以及血管生成因子旁分泌活性方面检测了c-Cbl下调对长期培养的MSCs的影响。此外，我们观察了长期培养的MSCs（有无c-Cbl下调）对糖尿病伤口的促愈合作用。我们发现，与第3代（P3）MSCs相比，第10代（P10）MSCs中c-Cbl的磷酸化水平升高，而Akt的磷酸化水平降低（P<0.05）。此外，P10 MSCs的增殖、旁分泌和迁移能力显著下降，同时细胞衰老增加（P<0.05）。然而，c-Cbl下调改善了P10 MSCs的这些功能，包括PI3K/Akt活性（P<0.05）。在糖尿病大鼠模型中，与P10 MSCs治疗相比，用c-Cbl下调的P10 MSCs治疗加速了糖尿病伤口愈合，表现为伤口闭合更快（P<0.05）、新生血管更多（P<0.05）以及伤口组织学评估得分更高（P<0.05）。我们的研究结果表明，c-Cbl下调可减轻长期培养诱导的MSCs促血管生成功能损伤，并改善长期培养的MSCs促进糖尿病伤口愈合的效果。
