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

作　　者：Shu-jie GUO Tai-rong WANG Jing CHEN Ling-yun WU Ping-jin GAO Ding-liang ZHU 

机构地区：[1]State Key Laboratory of Medical Genomics, Shanghai Key Laboratory of Vascular Biology and Department of Hypertension, Ruijin Hospital, Shanghai JiaoTong University School of Medicine, Shanghai 200025, China [2]Laboratory of Vascular Biology, Institute of Health Sciences, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences, Shanghai 200025, China [3]Shanghai Institute of Hypertension, Shanghai 200025, China [4]Luwan Branch, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China

出　　处：《Acta Pharmacologica Sinica》2010年第10期1312-1318,共7页中国药理学报（英文版）

摘　　要：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-as- sisted 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 immunofiuorescence 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 i were expressed in vascular AFs and their levels changed signifi- cantly 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.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-as- sisted 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 immunofiuorescence 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 i were expressed in vascular AFs and their levels changed signifi- cantly 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.

关 键 词：ADVENTITIA adventitial fibroblasts vascular remodeling spontaneously hypertensive rats two-dimensional electrophoresis MALDI-TOF-MS ANNEXIN 

分 类 号：Q463[生物学—生理学] Q51