Abstract
Salvianolic acid B (Sal B), a pure water-soluble compound extracted from Radix Salviae miltiorrhizae, has been reported to possess potential cardioprotective efficacy. To identify proteins or pathways by which Sal B might exert its protective activities on the cardiovascular system, two-dimensional gel electrophoresis-based comparative proteomics was performed, and proteins altered in their expression level after Sal B treatment were identified by MALDI-TOF MS/MS. Human umbilical vein endothelial cells (HUVECs) were incubated at Sal B concentrations that can be reached in human plasma by pharmacological intervention. Results indicated that caldesmon, an actin-stabilizing protein, was downregulated in Sal B-exposed HUVECs. Proteins that showed increased expression levels upon Sal B treatment were vimentin, T-complex protein 1, protein disulfide isomerase, tropomyosin alpha, heat shock protein beta-1, UBX domain-containing protein 1, alpha enolase, and peroxiredoxin-2. Additionally, Sal B leads to increased phosphorylation of nucleophosmin in a dose-dependent manner and promotes proliferation of HUVECs. We found that Sal B exhibited a coordinated regulation of enzymes and proteins involved in cytoskeletal reorganization, oxidative stress, and cell growth. Our investigation would provide understanding to the endothelium protection information of Sal B.
Highlights
Traditional Chinese medicine (TCM) has been widely practiced for thousands of years in China and Eastern Asia
Salvianolic acid B (Sal B), a pure water-soluble compound extracted from Danshen, has been reported to possess many biological activities attributed to the whole-plant Danshen herb, such as sedative, antioxidant, hepatoprotective and antifibrogenic effects [5], inhibition of platelet aggregation, improvement of coronary microcirculation and cerebral blood flow [6], enhancement of angiogenic processes, and protection against injury to the heart and brain caused by ischemia reperfusion [7]
human umbilical vein endothelial cells (HUVECs) treated with Sal B or not were washed twice with cold phosphate-buffered saline (PBS), lysed in PBS containing 1% Nonidet P-40, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate (SDS), 5 μg/mL aprotinin, 100 μg/mL phenylmethylsulfonyl fluoride, 1 μg/mL pepstatin A, and 1 mM ethylenediaminetetraacetic acid at 4◦C for 20 min, and disrupted with a needle
Summary
Traditional Chinese medicine (TCM) has been widely practiced for thousands of years in China and Eastern Asia. TCM is an integrated part of the healthcare system in that part of the world, and currently in China a strong push to modernize TCM through both scientific research and industrial development is underway. We Evidence-Based Complementary and Alternative Medicine aimed to investigate the modes of actions of Sal B for the prevention of vascular diseases. A previous study reported an HPLC method for the determination of Sal B in rat plasma after oral administration of Danshen extract and the pharmacokinetics of Sal B were investigated. To identify proteins or pathways by which Sal B might exert its protective activities on the cardiovascular system, we surveyed global changes in proteins after Sal B treatment in HUVECs. We report here our initial findings of Sal B on the protein expression in HUVECs based on the well-established 2D gel/MALDI-TOF/TOF mass spectrometer analyses. The differential expression of some proteins was confirmed by Western blot
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