Differential Proteomic Analysis of Platelets Suggested Possible Signal Cascades Network in Platelets Treated with Salvianolic Acid B

Abstract

BackgroundSalvianolic acid B (SB) is an active component isolated from Danshen, a traditional Chinese medicine widely used for the treatment of cardiovascular disorders. Previous study suggested that SB might inhibit adhesion as well as aggregation of platelets by a mechanism involving the integrin α2β1. But, the signal cascades in platelets after SB binding are still not clear.Methodology/Principal FindingsIn the present study, a differential proteomic analysis (two-dimensional electrophoresis) was conducted to check the protein expression profiles of rat platelets with or without treatment of SB. Proteins altered in level after SB exposure were identified by MALDI-TOF MS/MS. Treatment of SB caused regulation of 20 proteins such as heat shock-related 70 kDa protein 2 (hsp70), LIM domain protein CLP-36, copine I, peroxiredoxin-2, coronin-1 B and cytoplasmic dynein intermediate chain 2C. The regulation of SB on protein levels was confirmed by Western blotting. The signal cascades network induced by SB after its binding with integrin α2β1 was predicted. To certify the predicted network, binding affinity of SB to integrin α2β1 was checked in vitro and ex vivo in platelets. Furthermore, the effects of SB on protein levels of hsp70, coronin-1B and intracellular levels of Ca(2+) and reactive oxygen species (ROS) were checked with or without pre-treatment of platelets using antibody against integrin α2β1. Electron microscopy study confirmed that SB affected cytoskeleton structure of platelets.Conclusions/SignificanceIntegrin α2β1 might be one of the direct target proteins of SB in platelets. The signal cascades network of SB after binding with integrin α2β1 might include regulation of intracellular Ca(2+) level, cytoskeleton-related proteins such as coronin-1B and cytoskeleton structure of platelets.

Network Pharmacology: A New Approach for Chinese Herbal Medicine Research

“…Furthermore, nine components were screened in Fufang Danshen formula based on network pharmacology, which could modulate 42 cardiovascular-associated genes [25]. Additionally, by integrating the above research approaches, it was demonstrated that salvianolic acid B was compatible and feasible for cardiovascular disease treatment [26]. …”

Heat shock protein 70 regulates platelet integrin activation, granule secretion and aggregation

“…Chaperone proteins such as protein disulfide isomerase (PDI), ER oxidoreductin 1␣ (Ero1␣), ER protein 57 (ERp57), and ER protein 5 (ERp5) have emerging roles in regulating the activities of platelets and other hematopoietic cells (20,21,38,55). However, while both Hsp70 and Hsp90 family members have been identified in platelets (31,36,42,48), specific roles for Hsp70 activity in platelet function and aggregate formation have not been addressed.Here we investigate the role of Hsp70 in platelet function. We find that platelets express abundant Hsp70 protein and that Hsp70 localizes throughout platelets, while partially colocalizing with other chaperones such as Hsp90 and PDI.…”

Differential proteomic analysis of platelets suggested target-related proteins in rabbit platelets treated with Rhizoma Corydalis

“…Among various techniques used in platelet proteomics, gel-free and iTRAQ together with MS or LC-MS/MS, 2-DE coupled to kinds of MS were frequently applied to characterize platelets in cardiovascular diseases and in exploring effects of antiplatelet drugs (Coppinger et al 2007;Arias-Salgado et al 2008;Parguiña et al 2010Parguiña et al , 2011L opez-Farr e et al 2011). One of the application of 2-DE coupled to MS is to identify the expression of platelet proteins and suggest the possible signalling pathways after treated with traditional Chinese medicines (TCMs) and active compounds, such as Panax notoginseng (Burkill) F. H. Chen (Araliaceae) (Yao et al 2008b), salvianolic acids (Yao et al 2008a) and salvianolic acid B (Ma et al 2011) from Salvia miltiorrhiza Bunge (Labiatae).…”