miR-206对血管内皮细胞增殖的影响及其机制

目的研究miR-206对血管内皮细胞增殖的影响及其机制,为临床动脉硬化疾病提供新的诊疗方向。方法收集临床病理首次确诊为糖尿病下肢动脉粥样硬化病变的患者病变处动脉内皮组织和正常内皮组织,qPCR检测miR-206的表达;而后通过基础实验研究将HUVEC分为未转染组、miR-206-mimics组和miR-206-inhibitor组,观察转染效率;三组细胞均采用CCK8检测细胞增殖情况,Transwell进行迁移能力评估,流式细胞仪检测细胞周期情况,Western blot检测Cx43表达情况。结果糖尿病动脉硬化患者下肢动脉(均为胫腓动脉)病变处内皮细胞miR-206的mRNA表达较正常组织内皮细胞明显增加(P0.05);HUVEC转染miR-206-mimics后,miR-206表达显著升高(P0.05),转染miR-206-inhibitor后,miR-206表达明显降低(P0.05);在HUVEC转染miR-206-mimics 24 h后,细胞增殖率相比于未转染组和miR-206-inhibitor组要明显降低(P0.05);在HUVEC转染miR-206-mimics后,细胞迁移能力明显降低(P0.05),细胞周期在G2期发生阻滞,而HUVEC转染miR-206-inhibitor后细胞迁移能力提高(P0.05);miR-206-mimics组细胞Cx43表达明显降低,而miR-206-inhibitor组细胞Cx43表达明显增加。结论 miR-206可以抑制血管内皮细胞的增殖,将内皮细胞阻滞在有丝分裂G2期,并且抑制内皮细胞迁移,其对内皮细胞增殖的影响可能是通过调节Cx43表达来实现的。

Effect of miR-206 on proliferation of vascular endothelial cell and its mechanism

Aim To observe the effect of miR-206 on proliferation of vascular endothelial cell and its mechanism, and provide a new therapeutic target for clinical arteriosclerosis. Methods The arterial endothelial tissue on lesion site and normal endothelial tissue were collected from the patients who were diagnosed as diabetic lower limb atherosclerosis by first time in clinical pathology. The expression of miR-206 was detected by qPCR. Normal HUVEC cell lines, HUVEC transfected with miR-206-mimics and HUVEC transfected with miR-206-inhibitor were studied through fundamental experiment, and the transfection efficiency were observed. In the three groups, CCK8 was used to detect the proliferation status, transwell was used to evaluate the migration ability, flow cytometry was used to detect cell cycle and Western blot was used to observe expression of Cx43. Results The mRNA expression of miR-206 on lesion site of lower limb artery (shin peroneal artery) in the patients with diabetic arteriosclerosis was significantly increased compared with normal endothelial tissue(P<0.05); The expression of miR-206 significantly increased (P<0.05) after transfected with miR-206-mimics, while it was significantly reduced (P<0.05) after transfected with miR-206-inhibitor. Compared with normal HUVEC and HUVEC transfected with miR-206-inhibitor, the cell proliferation rate was significantly lower in HUVEC transfected with miR-206-mimics for 24 h (P<0.05). After HUVEC was transfected with miR-206-mimics, the cell migration capacity was significantly lower than normal HUVEC group (P<0.05), and the cell cycle was blocked during G2 phase; while the cell migration capacity was higher in HUVEC transfected with miR-206-inhibitor group than that of normal HUVEC group (P<0.05). Cx43 expression was significantly reduced in miR-206-mimics group but significantly increased in miR-206-inhibitor group. Conclusion miR-206 can inhibit the proliferation of vascular endothelial cells, block endothelial cells in G2 phase and suppress their migration, and it may affect proliferation of endothelial cells by regulating Cx43 expression.