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Differential proteomic analysis of platelets suggested target-related proteins in rabbit platelets treated with Rhizoma Corydalis
Li, Chun-Hong1; Chen, Cen2; Zhang, Qian1; Tan, Chen-Ning1; Hu, Yuan-Jia3; Li, Peng3; Wan, Jian-Bo3; Feng, Gang1; Xia, Zhi-Ning1; Yang, Feng-Qing1
2017
Source PublicationPHARMACEUTICAL BIOLOGY
ISSN1388-0209
Volume55Issue:1Pages:76-87
Abstract

Context: Corydalis yanhusuo W.T. Wang (Papaveraceae) (Rhizoma Corydalis) showed inhibitory effects on rabbit platelet aggregation induced by ADP, thrombin (THR) or arachidonic acid (AA).

Objective: This study separates and identifies the possible target-related platelet proteins and suggests possible signal cascades of RC antiplatelet aggregation.

Materials and methods: Based on comparative proteomics, the differentially expressed platelet proteins treated before and after with 50 mg/mL RC 90% ethanol extract (for 15 min at 37 °C) were analyzed and identified by two dimensional gel electrophoresis (2-DE) and MALDI-TOF-MS/MS. To further verify the possible signalling pathways of RC antiplatelet aggregation function, the concentration of calcium (Ca2+) was measured by Fura-2/AM fluorescence (Ex 340/380 nm, Em 500 nm) (RC final concentrations of 0.0156–0.1563 mg/mL), the levels of P-selectin and cyclic guanosine monophosphate (cGMP) were quantified by ELISA (OD. 450 nm) (RC final concentrations of 0.0156–1.5625 mg/mL), and the 5-hydroxytryptamine (5-HT) level was measured using ortho-phthalaldehyde (OPT) fluorescence (Ex 340 nm, Em 470 nm) (RC final concentrations of 0.3125–1.5625 mg/mL).

Results: The expression of 52 proteins were altered in rabbit platelets after the treatment and the MALDI-TOF-MS analysis indicated that those proteins include 12 cytoskeleton proteins, 7 cell signalling proteins, 3 molecular chaperone proteins, 6 proteins related to platelet function, 16 enzymes and 7 other related proteins. Furthermore, RC extract could decrease the levels of 5-HT [inhibition rate of 96.80% (p < 0.05, vs. THR-activated group) treated with 0.7813 mg/mL of RC], Ca2+ [172.73 ± 5.07 to 113.56 ± 5.46 nM (p < 0.001, vs. THR-activated group) treated with 0.0313 mg/mL of RC] and P-selectin [13.48 ± 0.96 ng/3 × 108 to 11.64 ± 0.17 ng/3 × 108 (p < 0.05, vs. THR-activated group) treated with 0.0156 mg/mL of RC], and increase in cGMP level [38.93 ± 0.57 to 50.26 ± 4.05 ng/3 × 108 (p < 0.05, vs. THR-activated group) treated with 1.5165 mg/mL of RC] in ADP (10 μmol/L), THR (0.25 u/mL) or AA-(0.205 mmol/L) activated rabbit platelets.

Discussion and conclusion: The present study indicated that P2Y12 receptor might be one of the direct target proteins of RC in platelets. The signal cascades network of RC after binding with P2Y12 receptor is mediating Gαi proteins to activate downstream signalling pathways (AC and/or PI3K signalling pathways) for the inhibition of platelet aggregation.

KeywordSignal Pathway Two Dimensional Electrophoresis Maldi-tof-ms Antiplatelet Aggregation
DOI10.1080/13880209.2016.1229340
URLView the original
Indexed BySCIE
WOS Research AreaPlant Sciences ; Medical Laboratory Technology ; Pharmacology & Pharmacy
WOS SubjectPlant Sciences ; Medical Laboratory Technology ; Pharmacology & Pharmacy
WOS IDWOS:000390846600088
PublisherTAYLOR & FRANCIS LTD
The Source to ArticleWOS
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Cited Times [WOS]:7   [WOS Record]     [Related Records in WOS]
Document TypeJournal article
CollectionInstitute of Chinese Medical Sciences
Corresponding AuthorYang, Feng-Qing
Affiliation1.School of Chemistry and Chemical Engineering, Chongqing University, Chongqing, China
2.Division of Imaging Science & Biomedical Engineering, King's College, London, UK
3.State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao, China
Recommended Citation
GB/T 7714
Li, Chun-Hong,Chen, Cen,Zhang, Qian,et al. Differential proteomic analysis of platelets suggested target-related proteins in rabbit platelets treated with Rhizoma Corydalis[J]. PHARMACEUTICAL BIOLOGY,2017,55(1):76-87.
APA Li, Chun-Hong.,Chen, Cen.,Zhang, Qian.,Tan, Chen-Ning.,Hu, Yuan-Jia.,...&Yang, Feng-Qing.(2017).Differential proteomic analysis of platelets suggested target-related proteins in rabbit platelets treated with Rhizoma Corydalis.PHARMACEUTICAL BIOLOGY,55(1),76-87.
MLA Li, Chun-Hong,et al."Differential proteomic analysis of platelets suggested target-related proteins in rabbit platelets treated with Rhizoma Corydalis".PHARMACEUTICAL BIOLOGY 55.1(2017):76-87.
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