Department of Pharmacology, School of Basic Medical Sciences, Peking University and Beijing Key Laboratory of Tumor Systems Biology, Peking University, Beijing, 100191, China.
Department of Wound Healing Center and Interventional Radiology and Vascular Surgery, Peking University Third Hospital, Beijing, 100191, China.
Acta Pharmacol Sin. 2024 Jul;45(7):1477-1491. doi: 10.1038/s41401-024-01246-3. Epub 2024 Mar 27.
Refractory wounds are a severe complication of diabetes mellitus that often leads to amputation because of the lack of effective treatments and therapeutic targets. The pathogenesis of refractory wounds is complex, involving many types of cells. Rho-associated protein kinase-1 (ROCK1) phosphorylates a series of substrates that trigger downstream signaling pathways, affecting multiple cellular processes, including cell migration, communication, and proliferation. The present study investigated the role of ROCK1 in diabetic wound healing and molecular mechanisms. Our results showed that ROCK1 expression significantly increased in wound granulation tissues in diabetic patients, streptozotocin (STZ)-induced diabetic mice, and db/db diabetic mice. Wound healing and blood perfusion were dose-dependently improved by the ROCK1 inhibitor fasudil in diabetic mice. In endothelial cells, fasudil and ROCK1 siRNA significantly elevated the phosphorylation of adenosine monophosphate-activated protein kinase at Thr172 (pThr-AMPKα), the activity of endothelial nitric oxide synthase (eNOS), and suppressed the levels of mitochondrial reactive oxygen species (mtROS) and nitrotyrosine formation. Experiments using integrated bioinformatics analysis and coimmunoprecipitation established that ROCK1 inhibited pThr-AMPKα by binding to receptor-interacting serine/threonine kinase 4 (RIPK4). These results suggest that fasudil accelerated wound repair and improved angiogenesis at least partially through the ROCK1/RIPK4/AMPK pathway. Fasudil may be a potential treatment for refractory wounds in diabetic patients.
难治性创面是糖尿病的一种严重并发症，由于缺乏有效治疗方法和治疗靶点，常导致截肢。难治性创面的发病机制复杂，涉及多种类型的细胞。Rho 相关蛋白激酶-1（ROCK1）可磷酸化一系列底物，触发下游信号通路，影响包括细胞迁移、通讯和增殖在内的多种细胞过程。本研究探讨了 ROCK1 在糖尿病创面愈合及分子机制中的作用。结果显示，糖尿病患者、链脲佐菌素（STZ）诱导的糖尿病小鼠和 db/db 糖尿病小鼠创面肉芽组织中 ROCK1 表达明显增加。ROCK1 抑制剂法舒地尔可剂量依赖性改善糖尿病小鼠的创面愈合和血流灌注。在内皮细胞中，法舒地尔和 ROCK1 siRNA 显著增加了 Thr172 位（pThr-AMPKα）的腺苷单磷酸激活蛋白激酶磷酸化、内皮型一氧化氮合酶（eNOS）的活性，并抑制了线粒体活性氧（mtROS）和硝基酪氨酸形成的水平。整合生物信息学分析和免疫共沉淀实验表明，ROCK1 通过与受体相互作用丝氨酸/苏氨酸激酶 4（RIPK4）结合抑制 pThr-AMPKα。这些结果表明，法舒地尔至少部分通过 ROCK1/RIPK4/AMPK 通路加速了创面修复和血管生成。法舒地尔可能是治疗糖尿病难治性创面的一种有潜力的药物。
