[OS-9与脯氨酰羟化酶在大鼠缺氧诱导性肺动脉高压中的协同作用]

[The cooperation of OS-9 and PHDs in hypoxia-induced pulmonary hypertension of rats].

作者信息

Pan Kun, Dai Ai-Guo, Hu Rui-Cheng

机构信息

Department of Respiratory Medicine, Hunan Institute of Gerontology, Hunan Province Geriatric Hospital, Changsha 410001, China.

出版信息

Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2009 Feb;25(1):1-6.

PMID:21186601

Abstract

AIM

To investigate the dynamic expression of hypoxia-inducible factor 1alpha, PHDs and OS-9 in pulmonary arteries of rats with hypoxia-induced pulmonary hypertension.

METHODS

SD rats were randomly divided into 5 groups (n = 8) and exposed to hypoxia for 0, 3, 7, 14 or 21 d, respectively. RT-PCR and in situ hybridization were used to determine the expression of mRNA. Immunohistochemistry and Western blot were used to determine the expression of protein.

RESULTS

HIF-1alpha protein was poorly positive in control, markedly up-regulated after 3 d and 7 d of hypoxia (P < 0.05, vs group C), and then declined slightly after 14 d and 21 d of hypoxia. HIF-1alpha mRNA increased dramatically after 14 d of hypoxia (P < 0.05, vs group C). PHD1, PHD2 mRNA and protein was positive in group C. PHD2 mRNA and protein were up-regulated after 3 d of hypoxia (P < 0.05, vs group C), reaching its peak after 14 d of hypoxia while PHD1 protein declined after 14 d of hypoxia (P < 0.05, vs group C) without statistic mRNA changing. PHD3 mRNA and protein were detected at low level in control, markedly up-regulated after 3 d of hypoxia (P < 0.05, vs group C), and then PHD3 mRNA kept at high level while PHD3 protein declined after 14 d of hypoxia (P < 0.05, vs 7 d). OS-9 mRNA was positively in control, markedly decreased after 3 d of hypoxia (P < 0.05, vs group C), reaching its lowest lever after 14 d of hypoxia. Linear correlation analysis showed that OS-9 protein was positively correlated with OS-9 mRNA (r = 0.82, P < 0.01) and HIF-1alpha protein (r = 0.57, P < 0.01).

CONCLUSION

HIF-1alpha, PHDs and OS-9 are all involved in the pathogenesis of hypoxic pulmonary hypertension in rats. OS-9 may interact with both HIF-1alpha and PHDs to promote PHD-mediated hydroxylation of HIF-1alpha.

摘要

目的

探讨缺氧诱导的肺动脉高压大鼠肺动脉中缺氧诱导因子1α（HIF-1α）、脯氨酰羟化酶（PHDs）和OS-9的动态表达。

方法

将SD大鼠随机分为5组（每组n = 8），分别缺氧0、3、7、14或21天。采用逆转录-聚合酶链反应（RT-PCR）和原位杂交法检测mRNA表达。采用免疫组织化学和蛋白质印迹法检测蛋白质表达。

结果

对照组HIF-1α蛋白弱阳性，缺氧3天和7天后明显上调（与C组相比，P < 0.05），缺氧14天和21天后略有下降。缺氧14天后HIF-1α mRNA显著增加（与C组相比，P < 0.05）。C组PHD1、PHD2 mRNA和蛋白呈阳性。缺氧3天后PHD2 mRNA和蛋白上调（与C组相比，P < 0.05），缺氧14天后达到峰值，而缺氧14天后PHD1蛋白下降（与C组相比，P < 0.05），mRNA无统计学变化。对照组PHD3 mRNA和蛋白表达水平较低，缺氧3天后明显上调（与C组相比，P < 0.05），之后PHD3 mRNA维持在高水平，而缺氧14天后PHD3蛋白下降（与7天相比，P < 0.05）。OS-9 mRNA在对照组呈阳性，缺氧3天后明显下降（与C组相比，P < 0.05），缺氧14天后降至最低水平。线性相关分析显示，OS-9蛋白与OS-9 mRNA（r = 0.82，P < 0.01）和HIF-1α蛋白（r = 0.57，P < 0.01）呈正相关。