Expression profiling of laser-microdissected intrapulmonary arteries in hypoxia-induced pulmonary hypertension.
Expression profiling of laser-microdissected intrapulmonary arteries in hypoxia-induced pulmonary hypertension.
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DOI：
10.1186/1465-9921-6-109
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发表时间：
2005-09-19
影响因子：
5.8
通讯作者：
Fink L
中科院分区：
文献类型：
--
作者：
Kwapiszewska G;Wilhelm J;Wolff S;Laumanns I;Koenig IR;Ziegler A;Seeger W;Bohle RM;Weissmann N;Fink L
慢性缺氧影响肺中的基因表达，导致肺动脉高压和血管重塑。对于血管室的具体调查，肺内动脉的激光显微解剖与阵列分析相结合。使用尼龙过滤器（1176个点）对缺氧1、7和21天（FiO 2 = 0.1）的小鼠进行分析。分别在第1、7和21天观察到29、38和42个基因表达的变化。基因根据其响应的时间过程分为5个不同的类别。通过芯片分析获得的基因调控通过实时PCR证实。此外，通过实时PCR测定生长介质PDGF-B、TGF-β、TSP-1、SRF、FGF-2、TIE-2受体和VEGF-R1的表达。在第1天，转录调节因子和离子相关蛋白质主要受到调节。然而，在第7天和第21天，观察到基质产生和降解基因的差异表达，表明正在进行的结构改变。在第1天上调的21个基因中，鉴定出15个基因携带缺氧诱导转录因子的潜在缺氧反应元件（HRE）。通过免疫组化检测3个差异表达基因（S100 A4、CD 36和FKBP 1a），证实其在蛋白水平上的调控。虽然FKBP 1a仅限于血管外膜，但S100 A4和CD 36定位于血管图尼卡中膜。激光显微切割和阵列分析揭示了几个新的基因参与肺血管重塑，以应对缺氧。免疫组织化学证实了三种蛋白质的调节，并指定其在血管平滑肌细胞和成纤维细胞中的定位，表明不同类型的细胞参与重塑过程。这种方法可以更深入地了解缺氧调节途径，特别是在这个复杂的器官的血管室。
Chronic hypoxia influences gene expression in the lung resulting in pulmonary hypertension and vascular remodelling. For specific investigation of the vascular compartment, laser-microdissection of intrapulmonary arteries was combined with array profiling. Analysis was performed on mice subjected to 1, 7 and 21 days of hypoxia (FiO2 = 0.1) using nylon filters (1176 spots). Changes in the expression of 29, 38, and 42 genes were observed at day 1, 7, and 21, respectively. Genes were grouped into 5 different classes based on their time course of response. Gene regulation obtained by array analysis was confirmed by real-time PCR. Additionally, the expression of the growth mediators PDGF-B, TGF-β, TSP-1, SRF, FGF-2, TIE-2 receptor, and VEGF-R1 were determined by real-time PCR. At day 1, transcription modulators and ion-related proteins were predominantly regulated. However, at day 7 and 21 differential expression of matrix producing and degrading genes was observed, indicating ongoing structural alterations. Among the 21 genes upregulated at day 1, 15 genes were identified carrying potential hypoxia response elements (HREs) for hypoxia-induced transcription factors. Three differentially expressed genes (S100A4, CD36 and FKBP1a) were examined by immunohistochemistry confirming the regulation on protein level. While FKBP1a was restricted to the vessel adventitia, S100A4 and CD36 were localised in the vascular tunica media. Laser-microdissection and array profiling has revealed several new genes involved in lung vascular remodelling in response to hypoxia. Immunohistochemistry confirmed regulation of three proteins and specified their localisation in vascular smooth muscle cells and fibroblasts indicating involvement of different cells types in the remodelling process. The approach allows deeper insight into hypoxic regulatory pathways specifically in the vascular compartment of this complex organ.
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影响因子：
82.9
作者：
Fink, L;Seeger, W;Bohle, RM
通讯作者：
Bohle, RM
影响因子：
4.6
作者：
Hoshikawa, Y;Nana-Sinkam, P;Geraci, MW
通讯作者：
Geraci, MW
DOI：
10.1164/ajrccm.158.6.9804076
发表时间：
1998-12-01
影响因子：
24.7
作者：
Berg, JT;Breen, EC;West, JB
通讯作者：
West, JB
影响因子：
7
作者：
Heid, CA;Stevens, J;Williams, PM
通讯作者：
Williams, PM
DOI：
10.1073/pnas.93.18.9493
发表时间：
1996-09-03
影响因子：
11.1
作者：
Hochachka, PW;Buck, LT;Land, SC
通讯作者：
Land, SC