维生素C调控肉鸡缺氧肺动脉血管平滑肌细胞缺氧诱导因子-1α mRNA转录的分子机制

Molecular Mechanism of Vitamin C on Hypoxia Inducible Factor-1α mRNA Transcription of Broiler Pulmonary Artery Smooth Muscle Cells under Hypoxia Condition

作者旨在通过脯氨酸羟化酶抑制剂(DMOG)和过氧化氢(H_2O_2)刺激肉鸡缺氧肺动脉平滑肌细胞(PASMCs)探讨维生素C(VC)对其氧化还原状态与缺氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)及其受体2(VEGFR2/Flk-1)mRNA转录调控的分子机制。在前期PASMCs培养和缺氧模型的基础上设计3个小试验,VC和H_2O_2、VC和DMOG、VC和H_2O_2+DMOG,每个试验5个处理,每个处理6个重复。结果表明:与常氧和缺氧空白组相比,试验1,VC显著增加SOD/MDA的比值(P<0.05),显著下调HIF-1α/VEGF/VEGFR2mRNA的转录水平(P<0.05),H_2O_2显著上调缺氧肉鸡PASMCs HIF-1αmRNA转录(P<0.05);试验2,DMOG显著上调SOD/MDA的比值(P<0.05),显著下调HIF-1α和VEGFR2mRNA转录(P<0.05),但显著上调VEGF mRNA的转录(P<0.05),VC+DMOG显著上调HIF-1α/VEGF/VEGFR2 mRNA转录(P<0.05);试验3,VC+H_2O_2+DMOG三者同时添加极显著增加HIF-1α/VEGF/VEGFR2 mRNA转录(P<0.01)。以上结果提示,VC能提升细胞外基质的抗氧化水平,其对缺氧基因表达的调控与细胞内脯氨酸羟化酶活性和氧化还原状态相关。

This study was conducted to investigate the molecular mechanism of Vitamin C （VC） on redox state and the mRNA transcription of hypoxia inducible factor-la（HIF-la）,Vascular en- dothelial growth factor （VEGF） and Vascular endothelial growth factor receptor 2 （VEGFR2/ Flk-1） in broiler PASMCs under hypoxia condition through dimethyloxalylglycine （DMOG） and H2 02. This study executed on the basis of the culture of PASMCs and hypoxia model,included 3 experiments,VC and H202 ,VC and DMOG,VC and H202-[-DMOG,including 5 treatments with six replicates of each experiment. Compared with control group under normoxic and hypoxic,Exp. 1 showed that VC significantly increased SOD to MDA ratio （P〈0.05） ,and down-regulated the mRNA transcription of HIF-la/VEGF/VEGFR2 （P〈0. 05） Exp. 2 showed that DMOG signif- icantly increased SOD to MDA ratio （P〈0. 05） ,and down-regulated the mRNA transcription of HIF-la and VEGFR2 （P〈0. 05） respectively, but significantly up-regulated the mRNA tran-scription of VEGF （P〈0. 05） ,VC ＋ DMOG significantly up-regulated the mRNA transcription of HIF-la/VEGF/VEGFR2 （P〈0. 05） ;Exp. 3 showed that VC-H202 ＋DMOG markedly up- regulated the mRNA transcription of HIF-lcdVEGF/VEGFR2 （P〈0. 01）. These results sugges- ted that VC could increase cellular antioxidantive ability, and regulate the relative expression of hypoxia genes based on the activity of proline hydroxylase and the redox state of broiler PASMCs.