FTO overexpression expedites wound healing and alleviates depression in burn rats through facilitating keratinocyte migration and angiogenesis via mediating TFPI-2 demethylation

• Published in: Molecular and cellular biochemistry Vol. 479; no. 2; pp. 325 - 335
• Main Authors: Xu, Zihan, Zhu, Xiumei, Mu, Shengzhi, Fan, Ronghui, Wang, Benfeng, Gao, Wenjie, Kang, Tao
• Format: Journal Article
• Language: English
• Published: New York Springer US 01.02.2024; Springer Nature B.V
• Subjects: Angiogenesis; Assaying; Biochemistry; Biological effects; Biomedical and Life Sciences; Body fat; Burns; Cancer Research; Cardiology; Cell migration; Cell proliferation; Cholecystokinin; Demethylation; Heat; Injuries; Keratinocytes; Life Sciences; Medical Biochemistry; Methylation; N6-methyladenosine; Plasmids; Proteins; siRNA; Tissue factor; Wound healing; FTO; Keratinocytes; Angiogenesis; Burn injury; TFPI-2
• ISSN: 0300-8177; 1573-4919
• DOI: 10.1007/s11010-023-04719-x

Burn injury is a serious traumatic injury that leads to severe physical and psychosocial impairment. Wound healing after burn injury is a substantial challenge in medical community. This study investigated the biological effects of the demethylase fat mass and obesity-associated protein (FTO) on burn injury. FTO protein level in burn skin tissues of patients was measured with Western blot assay. Keratinocytes (HaCaT cells) were given heat stimulation to induce an in vitro burn injury model, and then transfected with overexpression plasmids of FTO (pcDNA-FTO) or small interfering RNA against FTO (si-FTO). Cell proliferation, migration, and angiogenesis in keratinocytes were evaluated with CCK-8, Transwell, and tube formation assays, respectively. Tissue factor pathway inhibitor-2 (TFPI-2) m6A methylation level was detected with MeRIP‑qPCR assay. Then rescue experiments were conducted to explore the effects of FTO/TFPI-2 axis on keratinocyte functions. Lentivirus carrying FTO overexpression plasmids was injected into a burn rat model to detect its effects on wound healing and depressive-like behaviors in burn rats. FTO was downregulated in burn skin and heat-stimulated keratinocytes. FTO prominently augmented proliferation, migration and angiogenesis in heat-stimulated keratinocytes, while FTO knockdown showed the opposite results. FTO inhibited TFPI-2 expression by FTO-mediated m6A methylation modification. TFPI-2 overexpression abrogated FTO mediated enhancement of proliferation, migration and angiogenesis in keratinocytes. Additionally, FTO overexpression accelerated wound healing and improved depressive-like behaviors in burn rat model. FTO prominently augmented proliferation, migration and angiogenesis in heat-stimulated keratinocytes though inhibiting TFPI-2, and then improved wound healing and depressive-like behaviors.