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丙酮酸程序化預處理:脯氨酸羥化酶的差異性調控機制
來源: | 作者: | 發布時間:2014-4-15 17:22:14

丙酮酸程序化預處理:脯氨酸羥化酶的差異性調控機制
任昊,翟效月
(中國醫科大學,組織胚胎學教研室)
摘要:目的:開發一種新的丙酮酸預處理模擬低氧方法,保護視網膜免受光損傷。方法:6-8周齡的BALB / c小鼠丙酮酸處理。 Western印跡和實時PCR方法用于蛋白質和mRNA的研究。視網膜形態學染色檢查。 TUNEL染色和免費核試驗研究細胞凋亡。結果:首先,丙酮酸預處理使視網膜中缺氧誘導因子HIF- 1a高度表達。其次,PHD2水平同時增加。與此相反, HIF -2α和PHD1并不明顯受丙酮酸影響。 hemoxygenase -1和促紅細胞生成素( EPO )這兩個下游基因的表達起到了重要的保護作用。我們的數據揭示了PHD2 - HIF -1α的反饋回路,反應了丙酮酸上調HIF -1α的作用機制。丙酮酸預處理能夠在光損傷時減少視網膜細胞凋亡。結論:這種新的丙酮酸預處理方法的保護作用主要是由于HIF -1α的穩定(而不是HIF -2α)。這種誘導的差異是由于PHD的兩種亞型作為Hif的降解酶對丙酮酸的抑制作用不同而產生的。

關鍵詞:人體組織胚胎學; 缺氧誘導因子; 丙酮酸預處理;
中圖分類號:R329.1

A novel programmed application of pyruvate: differential stabilization of HIF-1a and HIF-2a
REN Hao, Xiaoyue Zhai
(china medical university, histology and embryology)
Foundations: Specialized Research Fund for the Doctoral Program of Higher Education (No. 20102104120020) Brief author introduction:associate prof.

Abstract: Purpose. To develop a novel hypoxia preconditioning mimicking approach by pyruvate programming in the retina focusing on the interplay between HIFs and prolyl hydroxylases (PHDs). Methods. 6-8 weeks old BALB/c mice was treated with pyruvate in a programmed regime. Western blotting and real time PCR were applied for the protein and mRNA studies. Retinal morphology was 10 checked by toluidine blue staining. TUNEL staining and free nucleosome assay were used to examine apoptosis. Retinal organic culture was developed to investigate the pyruvate mechanism in vitro. Result. As the predominant isomer in the retina, HIF-1a is highly stabilized by pyruvate both in vivo and vitro. This was followed by a concomitant increase of PHD2 level. In contrast, HIF-2a and PHD1 were not affected by pyruvate in vivo. The down-stream genes of hemoxygenase-1 (HO-1) and 15 erythropoietin (EPO) also mirrored the changes of the HIFs respectively. Our data revealed a PHD2-HIF-1a feedback loop, which was blocked by pyruvate and rendered further HIF-1a accumulation. In vitro, HIF-2a was also able to be stabilized by pyruvate inhibition to PHD1, but only happed after oxygen withdrawal. Pyruvate treatment reduced retinal apoptosis not only before but also after the light insult. Conclusion. This novel pyruvate programming was retinal protective not only with preconditioning but also post-conditioning. This protection was mainly due to HIF-1a stabilization but not HIF-2a. This differentiation was due to the specific abundance of PHD isoforms in the retina and importantly their distinct preference to HIFs as degradating enzymes.
Key words: histology and embryology; HIF; pyruvate pre-conditioning

 

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