Department of Vascular and Endovascular Surgery, The First Affiliated Hospital of Zhengzhou University, 1 East Jianshe Road, Zhengzhou, People's Republic of China.
Department of Vascular Surgery, Gongyi City People's Hospital, Gongyi, People's Republic of China.
Mol Med. 2023 Mar 28;29(1):39. doi: 10.1186/s10020-023-00635-4.
Diabetes-related limb ischemia is a challenge for lower extremity amputation. Aurora Kinase A (AURKA) is an essential serine/threonine kinase for mitosis, while its role in limb ischemia remains unclear.
Human microvascular endothelial cells (HMEC-1) were cultured in high glucose (HG, 25 mmol/L D-glucose) and no additional growth factors (ND) medium to mimic diabetes and low growth factors deprivation as in vitro model. Diabetic C57BL/6 mice were induced by streptozotocin (STZ) administration. After seven days, ischemia was surgically performed by left unilateral femoral artery ligation on diabetic mice. The vector of adenovirus was utilized to overexpress AURKA in vitro and in vivo.
In our study, HG and ND-mediated downregulation of AURKA impaired the cell cycle progression, proliferation, migration, and tube formation ability of HMEC-1, which were rescued by overexpressed AURKA. Increased expression of vascular endothelial growth factor A (VEGFA) induced by overexpressed AURKA were likely regulatory molecules that coordinate these events. Mice with AURKA overexpression exhibited improved angiogenesis in response to VEGF in Matrigel plug assay, with increased capillary density and hemoglobin content. In diabetic limb ischemia mice, AURKA overexpression rescued blood perfusion and motor deficits, accompanied by the recovery of gastrocnemius muscles observed by H&E staining and positive Desmin staining. Moreover, AURKA overexpression rescued diabetes-related impairment of angiogenesis, arteriogenesis, and functional recovery in the ischemic limb. Signal pathway results revealed that VEGFR2/PI3K/AKT pathway might be involved in AURKA triggered angiogenesis procedure. In addition, AURKA overexpression impeded oxidative stress and subsequent following lipid peroxidation both in vitro and in vivo, indicating another protective mechanism of AURKA function in diabetic limb ischemia. The changes in lipid peroxidation biomarkers (lipid ROS, GPX4, SLC7A11, ALOX5, and ASLC4) in in vitro and in vivo were suggestive of the possible involvement of ferroptosis and interaction between AUKRA and ferroptosis in diabetic limb ischemia, which need further investigation.
These results implicated a potent role of AURKA in diabetes-related impairment of ischemia-mediated angiogenesis and implied a potential therapeutic target for ischemic diseases of diabetes.
糖尿病相关的肢体缺血是下肢截肢的一个挑战。极光激酶 A（AURKA）是有丝分裂所必需的丝氨酸/苏氨酸激酶，但其在肢体缺血中的作用尚不清楚。
将人微血管内皮细胞（HMEC-1）在高糖（HG，25mmol/L D-葡萄糖）和无其他生长因子（ND）培养基中培养，以模拟糖尿病和低生长因子剥夺的体外模型。通过链脲佐菌素（STZ）给药诱导糖尿病 C57BL/6 小鼠。七天后，通过单侧左股动脉结扎对糖尿病小鼠进行手术缺血。利用腺病毒载体在体外和体内过表达 AURKA。
在我们的研究中，HG 和 ND 介导的 AURKA 下调抑制了 HMEC-1 的细胞周期进程、增殖、迁移和管形成能力，而过表达 AURKA 则挽救了这些作用。过表达 AURKA 诱导的血管内皮生长因子 A（VEGFA）的表达增加可能是协调这些事件的调节分子。在 Matrigel 塞植入物测定中，过表达 AURKA 的小鼠对 VEGF 表现出改善的血管生成，表现为毛细血管密度和血红蛋白含量增加。在糖尿病肢体缺血小鼠中，过表达 AURKA 挽救了血流灌注和运动功能障碍，同时通过 H&E 染色和阳性 Desmin 染色观察到比目鱼肌的恢复。此外，过表达 AURKA 挽救了糖尿病相关的血管生成、动脉生成和缺血肢体的功能恢复受损。信号通路结果表明，VEGFR2/PI3K/AKT 通路可能参与了 AURKA 触发的血管生成过程。此外，AURKA 过表达在体外和体内均抑制氧化应激和随后的脂质过氧化，表明 AURKA 在糖尿病肢体缺血中的另一种保护机制。体外和体内脂质过氧化生物标志物（脂质 ROS、GPX4、SLC7A11、ALOX5 和 ASLC4）的变化表明，在糖尿病肢体缺血中可能涉及铁死亡和 AUKRA 与铁死亡之间的相互作用，这需要进一步研究。
这些结果表明 AURKA 在糖尿病相关的缺血介导的血管生成受损中具有重要作用，并暗示了糖尿病缺血性疾病的潜在治疗靶点。
