【摘要】：目的：研究瘦素(Leptin)對臍靜脈內(nèi)皮細(xì)胞(HUVECs)增殖、遷移和管狀結(jié)構(gòu)形成能力的影響。并進(jìn)一步研究Leptin的促血管新生作用是否與之上調(diào)HIF-1α，進(jìn)而促進(jìn)下游靶基因VEGF表達(dá)有關(guān)及是否由PI3K and ERK1／2信號轉(zhuǎn)導(dǎo)通路所介導(dǎo)。 方法：分離、培養(yǎng)和鑒定HUVECs。檢測Leptin對HUVECs增殖、遷移和體外血管形成能力的影響。半定量RT-PCR檢測HIF-1α、VEGFmRNA的表達(dá)。Western blot檢測HIF-1α和VEGF蛋白的表達(dá)。 結(jié)果：Leptin能以劑量依賴性的方式促進(jìn)HUVECs增殖。Leptin100ng／mL時，HUVECs的遷移率明顯增加，是對照組的2.2倍，此作用能被PI3K抑制劑Ly294002和ERK1／2抑制劑PD98059所阻斷。Leptin作用后，HUVECs體外形成血管能力增強(qiáng)。Leptin呈劑量依賴性的上調(diào)HUVECS HIF-1α和VEGF蛋白表達(dá)，此作用依賴于Leptin誘導(dǎo)的HIF-1α和VEGF mRNA表達(dá)上調(diào)。缺氧模擬劑Cocl_2也能促進(jìn)HUVECs HIF-1α蛋白表達(dá)，，但對HIF-1αmRNA表達(dá)無影響，機(jī)制與Leptin不同。PI3K抑制劑Ly294002(20μmol／L)或ERK1／2抑制劑PD98059(20μmol／L)能夠抑制Leptin誘導(dǎo)的HIF-1α和VEGF蛋白表達(dá)上調(diào)，但對HIF-1α和VEGF mRNA表達(dá)無影響。 結(jié)論：Leptin通過促進(jìn)HUVECs增殖、遷移和體外血管形成等作用促進(jìn)血管新生。Leptin的促血管新生作用與之能夠在轉(zhuǎn)錄和翻譯水平上調(diào)HIF-1α表達(dá)，進(jìn)而促進(jìn)VEGF表達(dá)有關(guān)。PP3K和ERK1／2信號途徑
[Abstract]:Aim: to study the effects of leptin (Leptin) on proliferation, migration and tubular structure formation of umbilical vein endothelial cells (HUVECs). We further studied whether the angiogenesis-promoting effect of Leptin was related to the up-regulation of HIF-1 偽, and then to promote the expression of downstream target gene VEGF, and whether it was mediated by PI3K and ERK1/2 signal transduction pathway. Methods: HUVECs. was isolated, cultured and identified. The effects of Leptin on proliferation, migration and angiogenesis of HUVECs in vitro were detected. Semi-quantitative RT-PCR was used to detect the expression of HIF-1 偽 and VEGFmRNA. Western blot was used to detect the expression of HIF-1 偽 and VEGF protein. Results: Leptin promoted the proliferation of HUVECs in a dose-dependent manner. When Leptin 100 ng/mL, the migration rate of HUVECs increased significantly, which was 2.2 times higher than that of the control group. This effect could be blocked by PI3K inhibitor Ly294002 and ERK1/2 inhibitor PD98059. Leptin up-regulated the expression of HUVECS HIF-1 偽 and VEGF protein in a dose-dependent manner, which depended on the up-regulation of Leptin-induced expression of HIF-1 偽 and VEGF mRNA in HUVECs. Cocl_2, a hypoxia simulator, also promoted the expression of HUVECs HIF-1 偽 protein, but had no effect on the expression of HIF-1 偽 mRNA. PI3K inhibitor Ly294002 (20 渭 mol / L) or ERK1/2 inhibitor PD98059 (20 渭 mol / L) could inhibit the up-regulation of HIF-1 偽 and VEGF protein expression induced by Leptin, but had no effect on the expression of HIF-1 偽 and VEGF mRNA. Conclusion: Leptin promotes angiogenesis by promoting HUVECs proliferation, migration and angiogenesis in vitro. Leptin can promote angiogenesis by up-regulating the expression of HIF-1 偽 at transcription and translation levels. To promote the expression of VEGF. PP3K and ERK1/2 signaling pathway
【學(xué)位授予單位】：中國協(xié)和醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2006
【分類號】：R33

【引證文獻(xiàn)】

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1 王春艷;杜瑞平;張興夫;高民;;瘦素及其生理功能概述[J];動物營養(yǎng)學(xué)報;2012年03期

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