Wenzhou Municipal Key Cardiovascular Research Laboratory, Department of Cardiology, The First Affiliated Hospital, Wenzhou Medical University, Wenzhou 325000, Zhejiang Province, China.
Provincial Key Cardiovascular Research Laboratory, Department of Cardiology, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310009, Zhejiang Province, China.
EBioMedicine. 2020 Feb;52:102637. doi: 10.1016/j.ebiom.2020.102637. Epub 2020 Jan 22.
Canagliflozin (CANA) administration increases the risk of lower limb amputation in the clinic. The present study aimed to investigate whether and how CANA interferes with the intracellular physiological processes of bone marrow derived mesenchymal stem cells (BM-MSCs) and its contribution to ischaemic lower limb.
The in vivo blood flow recovery in ischaemic lower limbs following CANA treatment was evaluated. The cellular function of BM-MSCs after CANA treatment were also assessed in vitro. In silico docking analysis and mutant substitution assay were conducted to confirm the interaction of CANA with glutamate dehydrogenase 1 (GDH1).
Following CANA treatment, attenuated angiogenesis and hampered blood flow recovery in the ischaemic region were detected in diabetic and non-diabetic mice, and inhibition of the proliferation and migration of BM-MSCs were also observed. CANA was involved in mitochondrial respiratory malfunction in BM-MSCs and the inhibition of ATP production, cytochrome c release and vessel endothelial growth factor A (VEGFA) secretion, which may contribute to reductions in the tissue repair capacity of BM-MSCs. The detrimental effects of CANA on MSCs result from the inhibition of GDH1 by CANA (evidenced by in silico docking analysis and H199A-GDH1/N392A-GDH1 mutant substitution).
Our work highlights that the inhibition of GDH1 activity by CANA interferes with the metabolic activity of the mitochondria, and this interference deteriorates the retention of and VEGFA secretion by MSCs.
National Natural Science Foundation of China, Natural Science Foundation of Zhejiang Province and Wenzhou Science and Technology Bureau Foundation.
卡格列净（CANA）的应用会增加临床下肢截肢的风险。本研究旨在探讨卡格列净是否以及如何干扰骨髓间充质干细胞（BM-MSCs）的细胞内生理过程，并对缺血性下肢产生影响。
评估卡格列净治疗后缺血性下肢的体内血流恢复情况。还在体外评估了卡格列净处理后 BM-MSCs 的细胞功能。通过计算机对接分析和突变取代试验，证实了卡格列净与谷氨酸脱氢酶 1（GDH1）的相互作用。
在糖尿病和非糖尿病小鼠中，卡格列净治疗后，缺血区域的血管生成减弱，血流恢复受阻，BM-MSCs 的增殖和迁移受到抑制。卡格列净参与了 BM-MSCs 线粒体呼吸功能障碍，抑制了 ATP 产生、细胞色素 c 释放和血管内皮生长因子 A（VEGFA）分泌，这可能导致 BM-MSCs 的组织修复能力下降。卡格列净对间充质干细胞的有害影响源于卡格列净对 GDH1 的抑制（通过计算机对接分析和 H199A-GDH1/N392A-GDH1 突变取代证实）。
我们的工作强调了卡格列净抑制 GDH1 活性会干扰线粒体的代谢活性，这种干扰会降低间充质干细胞的保留和 VEGFA 的分泌。
国家自然科学基金、浙江省自然科学基金和温州市科技局基金。
