Qiu Tie-Ying, Huang Jin, Wang Li-Ping, Zhu Bi-Song
Clinical Nursing Teaching and Research Section of the Second Xiangya Hospital, Changsha 410011, P.R. China.
Organ Transplant Center, Xiangya Hospital, Central South University, Changsha 410008, P.R. China. Emails:
Cell J. 2021 Apr;23(1):51-60. doi: 10.22074/cellj.2021.7080. Epub 2021 Mar 1.
Patients with diabetes mellitus frequently have chronic wounds or diabetic ulcers as a result of impaired wound healing, which may lead to limb amputation. Human umbilical vein endothelial cell (HUVEC) dysfunction also delays wound healing. Here, we investigated the mechanism of miR-200b in HUVECs under high glucose conditions and the potential of miR-200b as a therapeutic target.
In this experimental study, HUVECs were cultured with 5 or 30 mM glucose for 48 hours. Cell proliferation was evaluated by CCK-8 assays. Cell mobility was tested by wound healing and Transwell assays. Angiogenesis was analyzed Matrigel tube formation assays. Luciferase reporter assays were used to test the binding of miR-200b with Notch1.
miR-200b expression was induced by high glucose treatment of HUVECs (P<0.01), and it significantly repressed cell proliferation, migration, and tube formation (P<0.05). Notch1 was directly targeted and repressed by miR-200b at both the mRNA and protein levels. Inhibition of miR-200b restored Notch1 expression (P<0.05) and reactivated the Notch pathway. The effects of miR-200b inhibition in HUVECs could be reversed by treatment with a Notch pathway inhibitor (P<0.05), indicating that the miR-200b/Notch axis modulates the proliferation, migration, and tube formation ability of HUVECs.
Inhibition of miR-200b activated the angiogenic ability of endothelial cells and promoted wound healing through reactivation of the Notch pathway . miR-200b could be a promising therapeutic target for treating HUVEC dysfunction.
糖尿病患者常因伤口愈合受损而出现慢性伤口或糖尿病溃疡，这可能导致肢体截肢。人脐静脉内皮细胞（HUVEC）功能障碍也会延迟伤口愈合。在此，我们研究了高糖条件下miR-200b在HUVEC中的作用机制以及miR-200b作为治疗靶点的潜力。
在本实验研究中，将HUVEC分别用5或30 mM葡萄糖培养48小时。通过CCK-8法评估细胞增殖。通过伤口愈合实验和Transwell实验检测细胞迁移能力。用基质胶管形成实验分析血管生成。用荧光素酶报告基因实验检测miR-200b与Notch1的结合。
高糖处理HUVEC可诱导miR-200b表达（P<0.01），且其显著抑制细胞增殖、迁移和管形成（P<0.05）。miR-200b在mRNA和蛋白水平直接靶向并抑制Notch1。抑制miR-200b可恢复Notch1表达（P<0.05）并重新激活Notch信号通路。用Notch信号通路抑制剂处理可逆转HUVEC中miR-200b抑制的作用（P<0.05），表明miR-200b/Notch轴调节HUVEC的增殖、迁移和管形成能力。
抑制miR-200b可激活内皮细胞的血管生成能力，并通过重新激活Notch信号通路促进伤口愈合。miR-200b可能是治疗HUVEC功能障碍的一个有前景的治疗靶点。
