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目的:研究瘦素(leptin)对脐静脉内皮细胞(HUVECs)增殖、迁移和管状结构形成能力的影响。方法:半定量RT-PCR检测HIF-1α、VEGFmRNA的表达,蛋白质印迹法检测HIF-1α和VEGF蛋白的表达。结果:10ng/mLleptin呈现促增殖效应,OD值为0.572±0.027,明显高于对照组的0.284±0.031(P<0.05);10ng/mL瘦素迁移率为(21.1±0.32)%,明显高于对照组的(9.99±0.43)%,P<0.05;管状结构的总长度为(5.8±0.5)mm,对照组无管状结构形成,P<0.05。瘦素呈剂量依赖性的上调HU-VECsHIF-1α和VEGF蛋白表达,最大诱导效应发生在1μg/mL,对照组不表达;HIF-1α和VEGFmRNA上调直接导致HIF-1α和VEGF蛋白合成增加,对照组的缺氧模拟剂CoCl2对HIF-1αmRNA表达无影响。结论:瘦素通过促进HUVECs增殖、迁移和体外血管形成等作用促进血管新生。瘦素的促血管新生作用能够在转录和翻译水平上调HIF-1α表达,进而转录激活VEGF表达有关。
Objective: To study the effect of leptin on the proliferation, migration and tubular structure formation of human umbilical vein endothelial cells (HUVECs). Methods: The expression of HIF-1α and VEGF mRNA was detected by semi-quantitative RT-PCR and the expressions of HIF-1α and VEGF protein were detected by Western blotting. Results: 10ng / mL leptin showed a proliferative effect with an OD of 0.572 ± 0.027, which was significantly higher than that of the control group (0.284 ± 0.031, P <0.05). The migration rate of 10ng / mL leptin was (21.1 ± 0.32)%, The control group (9.99 ± 0.43)%, P <0.05; the total length of the tubular structure was (5.8 ± 0.5) mm, the control group had no tubular structure, P <0.05. Leptin up-regulated the expression of HIF-1α and VEGF protein in a dose-dependent manner. The maximum induction rate was 1μg / mL in HU-VECs. The up-regulation of HIF-1α and VEGF mRNA directly induced the increase of HIF-1α and VEGF protein. Group hypoxia simulator CoCl2 had no effect on HIF-1αmRNA expression. Conclusion: Leptin promotes angiogenesis by promoting the proliferation, migration and extracorporeal vascularization of HUVECs. The pro-angiogenic effect of leptin can be related to the up-regulation of HIF-1α expression at transcriptional and translational levels, which in turn leads to transcriptional activation of VEGF expression.