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Differential proteomic analysis of platelets suggested possible signal cascades network in platelets treated with salvianolic acid B.

Ma C, Yao Y, Yue QX, Zhou XW, Yang PY, Wu WY, Guan SH, Jiang BH, Yang M, Liu X, Guo DA - PLoS ONE (2011)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Research Center for Modernization of Traditional Chinese Medicine, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, People's Republic of China.

ABSTRACT

Background: Salvianolic 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 findings: In 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 Ca²+ 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/significance: Integrin 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 Ca²+ level, cytoskeleton-related proteins such as coronin-1B and cytoskeleton structure of platelets.

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The proteome maps of control and SB-treated platelets.A, Representative two-dimensional electrophoresis gel pair images. Differentially expressed spots were shown by the arrows. B, The expanded region of differentially expressed protein spots in A. The proteins within the circles were the differentially expressed proteins.
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pone-0014692-g002: The proteome maps of control and SB-treated platelets.A, Representative two-dimensional electrophoresis gel pair images. Differentially expressed spots were shown by the arrows. B, The expanded region of differentially expressed protein spots in A. The proteins within the circles were the differentially expressed proteins.

Mentions: Representative two-dimensional electrophoresis gel images for control, SB-treated platelets were shown in A of Fig. 2. Fifteen up-regulated protein spots and five down-regulated protein spots was found as indicated by the arrowed spots in A of Fig. 2 and by the expanded plots in B of Fig. 2. Table S1 showed the average intensity values and their standard deviations of the spots and the fold differences between control and SB-treated group. The fold difference was represented by the ratio of the intensity value of SB-treated group to the value of control group. MS/MS analysis results of the differentially expressed proteins were also summarized in Table S1. The protein score, coverage and best ion score of each protein were showed. The result of MALDI-TOF MS/MS analysis of spot 4 was shown in Fig. 3 as an example.


Differential proteomic analysis of platelets suggested possible signal cascades network in platelets treated with salvianolic acid B.

Ma C, Yao Y, Yue QX, Zhou XW, Yang PY, Wu WY, Guan SH, Jiang BH, Yang M, Liu X, Guo DA - PLoS ONE (2011)

The proteome maps of control and SB-treated platelets.A, Representative two-dimensional electrophoresis gel pair images. Differentially expressed spots were shown by the arrows. B, The expanded region of differentially expressed protein spots in A. The proteins within the circles were the differentially expressed proteins.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3040754&req=5

pone-0014692-g002: The proteome maps of control and SB-treated platelets.A, Representative two-dimensional electrophoresis gel pair images. Differentially expressed spots were shown by the arrows. B, The expanded region of differentially expressed protein spots in A. The proteins within the circles were the differentially expressed proteins.
Mentions: Representative two-dimensional electrophoresis gel images for control, SB-treated platelets were shown in A of Fig. 2. Fifteen up-regulated protein spots and five down-regulated protein spots was found as indicated by the arrowed spots in A of Fig. 2 and by the expanded plots in B of Fig. 2. Table S1 showed the average intensity values and their standard deviations of the spots and the fold differences between control and SB-treated group. The fold difference was represented by the ratio of the intensity value of SB-treated group to the value of control group. MS/MS analysis results of the differentially expressed proteins were also summarized in Table S1. The protein score, coverage and best ion score of each protein were showed. The result of MALDI-TOF MS/MS analysis of spot 4 was shown in Fig. 3 as an example.

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Research Center for Modernization of Traditional Chinese Medicine, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, People's Republic of China.

ABSTRACT

Background: Salvianolic 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 findings: In 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 Ca²+ 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/significance: Integrin 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 Ca²+ level, cytoskeleton-related proteins such as coronin-1B and cytoskeleton structure of platelets.

Show MeSH
Related in: MedlinePlus