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Dynamic expression of proteins associated with adventitial remodeling in adventitial fibroblasts from spontaneously hypertensive rats.

Guo SJ, Wang TR, Chen J, Wu LY, Gao PJ, Zhu DL - Acta Pharmacol. Sin. (2010)

Bottom Line: Except for cytoskeleton proteins such as tubulin beta 5, it was found that annexin A1, translation elongation factor Tu, endoplasmic reticulum protein 29 and calcium-binding protein 1 were expressed in vascular AFs and their levels changed significantly in SHR-AFs compared with those in WKY-AFs.A decrease in annexin A1 in SHR-AFs was confirmed with Western blotting and immunofluorescence staining at the cell and tissue levels.The application of proteomic techniques revealed a number of novel proteins involved in adventitial remodeling of AFs from SHR, which provide new mechanisms responsible for the occurrence and development of hypertension and potential targets for influencing vascular remodeling in hypertension.

View Article: PubMed Central - PubMed

Affiliation: Department of Hypertension, Ruijin Hospital, Shanghai JiaoTong University School of Medicine, China.

ABSTRACT

Aim: To identify proteins that could potentially be involved in adventitial remodeling in vascular adventitial fibroblasts (AFs) from spontaneously hypertensive rats (SHR).

Methods: AFs were isolated from thoracic aortas of 4-, 8-, 16-, and 24-week-old male SHR and Wistar-Kyoto (WKY) rats and cultured to passage 4. Proteomic differential expression profiles between SHR-AFs and WKY-AFs were investigated using 2-D electrophoresis (2-DE), whereas gel image analysis was processed using Image Master 2D Platinum. Protein spots were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Expression levels of annexin A1 in AFs and aortas from SHR and WKY rats were detected with Western blotting and immunofluorescence techniques.

Results: In 4-, 8-, 16-, and 24-week-old SHR-AFs, 49, 59, 54, and 69 protein spots were found to have significant differences from the age-matched WKY-AFs. Fourteen spots with the same changes in patterns were analyzed in 4-, 8-, 16-, and 24-week-old SHR-AFs with mass spectrometry. Except for cytoskeleton proteins such as tubulin beta 5, it was found that annexin A1, translation elongation factor Tu, endoplasmic reticulum protein 29 and calcium-binding protein 1 were expressed in vascular AFs and their levels changed significantly in SHR-AFs compared with those in WKY-AFs. A decrease in annexin A1 in SHR-AFs was confirmed with Western blotting and immunofluorescence staining at the cell and tissue levels.

Conclusion: The application of proteomic techniques revealed a number of novel proteins involved in adventitial remodeling of AFs from SHR, which provide new mechanisms responsible for the occurrence and development of hypertension and potential targets for influencing vascular remodeling in hypertension.

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Altered location of spot 13 in 2-DE maps of vascular adventitia fibroblasts from SHR and WKY rats.
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fig2: Altered location of spot 13 in 2-DE maps of vascular adventitia fibroblasts from SHR and WKY rats.

Mentions: To explore the differences in protein expression between SHR-AFs and WKY-AFs, the proteins from SHR-AFs and WKY-AFs from 4-, 8-, 16-, and 24-week-old mice were separated by 2-DE. Gels using three different samples from the same group were performed simultaneously and analyzed by ImageMaster 2D Platinum Analysis Software. In total, 1228±132 spots were detected on the maps, and the overall protein expression profiles with pH 3−10 and molecular masses of 10 to 90 kDa were very similar within the three samples of the same group as analyzed by ImageMaster 2D Platinum, indicating high stability and reproducibility of the 2-DE in our test system. In comparison with the 2-DE maps of age-matched WKY-AFs, the number of altered spots, up-regulated spots, down-regulated spots and spots with changed locations for SHR-AFs were, respectively, as follows: 4-week-old rats, 49, 29, 19, and 1 spots; 8-week-old rats, 59, 34, 23, and 2 spots; 16-week-old rats, 54, 33, 20, and 1; and 24-week-old rats, 69, 42, 25, and 2. Thirteen protein spots with significant changes in volume (paired t-test, P<0.05) in a consistent direction (increased or decreased in all three samples per group and in four age groups) and one spot with an altered location were judged as differential spots and excised for identification as noted in Figures 1 and 2.


Dynamic expression of proteins associated with adventitial remodeling in adventitial fibroblasts from spontaneously hypertensive rats.

Guo SJ, Wang TR, Chen J, Wu LY, Gao PJ, Zhu DL - Acta Pharmacol. Sin. (2010)

Altered location of spot 13 in 2-DE maps of vascular adventitia fibroblasts from SHR and WKY rats.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4012903&req=5

fig2: Altered location of spot 13 in 2-DE maps of vascular adventitia fibroblasts from SHR and WKY rats.
Mentions: To explore the differences in protein expression between SHR-AFs and WKY-AFs, the proteins from SHR-AFs and WKY-AFs from 4-, 8-, 16-, and 24-week-old mice were separated by 2-DE. Gels using three different samples from the same group were performed simultaneously and analyzed by ImageMaster 2D Platinum Analysis Software. In total, 1228±132 spots were detected on the maps, and the overall protein expression profiles with pH 3−10 and molecular masses of 10 to 90 kDa were very similar within the three samples of the same group as analyzed by ImageMaster 2D Platinum, indicating high stability and reproducibility of the 2-DE in our test system. In comparison with the 2-DE maps of age-matched WKY-AFs, the number of altered spots, up-regulated spots, down-regulated spots and spots with changed locations for SHR-AFs were, respectively, as follows: 4-week-old rats, 49, 29, 19, and 1 spots; 8-week-old rats, 59, 34, 23, and 2 spots; 16-week-old rats, 54, 33, 20, and 1; and 24-week-old rats, 69, 42, 25, and 2. Thirteen protein spots with significant changes in volume (paired t-test, P<0.05) in a consistent direction (increased or decreased in all three samples per group and in four age groups) and one spot with an altered location were judged as differential spots and excised for identification as noted in Figures 1 and 2.

Bottom Line: Except for cytoskeleton proteins such as tubulin beta 5, it was found that annexin A1, translation elongation factor Tu, endoplasmic reticulum protein 29 and calcium-binding protein 1 were expressed in vascular AFs and their levels changed significantly in SHR-AFs compared with those in WKY-AFs.A decrease in annexin A1 in SHR-AFs was confirmed with Western blotting and immunofluorescence staining at the cell and tissue levels.The application of proteomic techniques revealed a number of novel proteins involved in adventitial remodeling of AFs from SHR, which provide new mechanisms responsible for the occurrence and development of hypertension and potential targets for influencing vascular remodeling in hypertension.

View Article: PubMed Central - PubMed

Affiliation: Department of Hypertension, Ruijin Hospital, Shanghai JiaoTong University School of Medicine, China.

ABSTRACT

Aim: To identify proteins that could potentially be involved in adventitial remodeling in vascular adventitial fibroblasts (AFs) from spontaneously hypertensive rats (SHR).

Methods: AFs were isolated from thoracic aortas of 4-, 8-, 16-, and 24-week-old male SHR and Wistar-Kyoto (WKY) rats and cultured to passage 4. Proteomic differential expression profiles between SHR-AFs and WKY-AFs were investigated using 2-D electrophoresis (2-DE), whereas gel image analysis was processed using Image Master 2D Platinum. Protein spots were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Expression levels of annexin A1 in AFs and aortas from SHR and WKY rats were detected with Western blotting and immunofluorescence techniques.

Results: In 4-, 8-, 16-, and 24-week-old SHR-AFs, 49, 59, 54, and 69 protein spots were found to have significant differences from the age-matched WKY-AFs. Fourteen spots with the same changes in patterns were analyzed in 4-, 8-, 16-, and 24-week-old SHR-AFs with mass spectrometry. Except for cytoskeleton proteins such as tubulin beta 5, it was found that annexin A1, translation elongation factor Tu, endoplasmic reticulum protein 29 and calcium-binding protein 1 were expressed in vascular AFs and their levels changed significantly in SHR-AFs compared with those in WKY-AFs. A decrease in annexin A1 in SHR-AFs was confirmed with Western blotting and immunofluorescence staining at the cell and tissue levels.

Conclusion: The application of proteomic techniques revealed a number of novel proteins involved in adventitial remodeling of AFs from SHR, which provide new mechanisms responsible for the occurrence and development of hypertension and potential targets for influencing vascular remodeling in hypertension.

Show MeSH
Related in: MedlinePlus