Vasoactive peptides upregulate mRNA expression and secretion of vascular endothelial growth factor in human airway smooth muscle cells
Vasoactive peptides upregulate mRNA expression and secretion of vascular endothelial growth factor in human airway smooth muscle cells
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DOI：
10.1385/cbb:47:1:109
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发表时间：
2007-01-01
影响因子：
2.6
通讯作者：
Sharma, Hari S.
中科院分区：
文献类型：
--
作者：
Alagappan, Vijay K. T.;Willems-Widyastuti, Anna;Sharma, Hari S.
气道重塑和相关的血管生成是哮喘的特征，但其分子机制尚未完全了解。血管紧张素 (ANG)II 和内皮素 (ET)-1 是与哮喘有关的有效血管收缩循环激素。我们研究了 ANG 11 和 ET-1 对人气道平滑肌 (ASM) 细胞增殖和生长的影响，并检查了血管生成肽、血管内皮生长因子 (VEGF) 的 mRNA 表达和释放。将去血清（48 小时）的人 ASM 细胞与 ANG 11 (100 nM) 或 ET-1 (10 nM) 一起孵育 30 分钟、1、2、4、8、16 和 24 小时，并检查 VEGF 的内源合成与接受无血清培养基的对照细胞的关系。通过 [H-3]-胸苷掺入测定确定 ET-1 诱导时间依赖性 DNA 生物合成。使用Northern印迹杂交，我们在培养的平滑肌细胞中检测到编码VEGF的两种3.9和1.7kb的mRNA种类。 ANG 11 和 ET-1 均诱导 VEGF 的 mRNA 表达（2 至 3 倍）和分泌（1.8 至 2.8 倍）在孵育 4-8 小时内达到最大水平。 ANG 11 孵育 8 小时后，诱导的 VEGF 表达和释放下降，而 ET-1 的水平仍然升高。源自 ET-1 处理的 ASM 细胞的条件培养基诱导猪肺动脉内皮细胞和人脐静脉内皮细胞中的 [H-3]-胸苷掺入和细胞数量。此外，VEGF酪氨酸激酶受体抑制剂阻断了条件培养基诱导的内皮细胞有丝分裂发生。我们的结果表明，ANG II 和 ET-1 在 ASM 细胞生长和 VEGF 上调中具有潜在作用，VEGF 可能通过旁分泌机制参与内皮细胞增殖，从而导致哮喘期间出现的病理性血管生成和血管重塑。
Airway remodeling and associated angiogenesis are documented features of asthma, of which the molecular mechanisms are not fully understood. Angiotensin (ANG)II and endothelin (ET)-1 are potent vasoconstricting circulatory hormones implicated in asthma. We investigated the effects of ANG 11 and ET-1 on human airway smooth muscle (ASM) cells proliferation and growth and examined the mRNA expression and release of the angiogenic peptide, vascular endothelial growth factor (VEGF). Serum deprived (48 h) human ASM cells were incubated with ANG 11 (100 nM) or ET-1 (10 nM) for 30 min, 1, 2, 4, 8, 16, and 24 h and the endogenous synthesis of VEGF was examined in relation to control cells receiving serum free culture medium. ET-1 induced time dependent DNA biosynthesis as determined by [H-3]-thymidine incorporation assay. Using northern blot hybridization, we detected two mRNA species of 3.9 and 1.7 kb encoding VEGF in the cultured smooth muscle cells. Both ANG 11 and ET-1 induced the mRNA expression (two- to threefold) and secretion (1.8- to 2.8-fold) of VEGF reaching maximal levels between 4-8 h of incubation. Induced expression and release of VEGF declined after 8 h of ANG 11 incubation while levels remained elevated in the case of ET-1. The conditioned medium derived from ET-1-treated ASM cells induced [H-3]-thymidine incorporation and cell number in porcine pulmonary artery endothelial as well as human umbilical vein endothelial cells. Moreover, the VEGF tyrosine kinase receptor inhibitor blocked the conditioned medium induced mitogenesis in endothelial cells. Our results suggest a potential role for ANG II and ET-1 in ASM cell growth and upregulation of VEGF that may participate in endothelial cell proliferation via paracrine mechanisms and thus causing pathological angiogenesis and vascular remodelling seen during asthma.