title: Tanshinone IIA, a major component of Salvia milthorriza Bunge, inhibits platelet activation via Erk-2 signaling pathway.
abstract: BACKGROUND
  The roots of Salvia milthorriza Bunge (Lamiaceae) known as "Danshen", are used in Traditional Chinese Medicine as a remedy for activating blood and eliminating stasis. TIIA, a diterpenoid of Salvia milthorriza, is one of active components in Danshen that exhibits a significant improvement of the blood flow in the coronary circulatory system and a reduction of myocardial infarction. However, its effect on platelet and underlying mechanism remains largely unknown. On this basis, this compound could be a promising agent to improve blood viscosity and microcirculation and to prevent CVD.
  
  
  METHODS
  In order to investigate the effects of TIIA on platelet functionality and its interaction with various platelet activation pathways, rat PRP were incubated with TIIA for 1 min at 37°C prior the addition of the stimuli (ADP or collagen). Aggregation was monitored in a light transmission aggregometer measuring changes in turbidity with continuous observation up to 10 min after the addition of the stimuli. MAPK signaling pathway and tubulin acetylation were analyzed by a Western blot technique. The effect of the TIIA was also studied in vivo on bleeding time in mice.
  
  
  RESULTS
  TIIA selectively inhibited rat platelet aggregation induced by reversible ADP stimuli (3 μM) in a concentration-dependent manner (0.5-50 μM). Nevertheless, TIIA was less active against the irreversible stimuli induced by ADP (10 μM) and collagen (10 μg/mL). Moreover, experiments performed on platelet lysates collected at different time-point after the addition of the stimuli shown that TIIA modulated tubulin acetylation and inhibited Erk-2 phosphorylation. Concomitantly, TIIA administrated i.p. at 10 mg/kg significantly amplified the mice bleeding time with an increase of 58% compared to its control (2.06±0.29 min vs 1.30±0.07). ASA was used as reference drug for in vitro and in vivo experiments.
  
  
  CONCLUSIONS
  This study clarifies the intracellular signaling pathway involved in antiplatelet action of TIIA and also gives preliminary evidences for its anticoagulant activity. On this basis, this compound could be a promising agent to improve blood viscosity and microcirculation and to prevent CVD.
authors.affiliation: Department of Pharmacy, University of Naples Federico II, Via Domenico Montesano 49, 80131 Naples, Italy. Electronic address: francesco.maione@unina.it.
authors.forename: Francesco
authors.initials: F
authors.lastname: Maione
authors.affiliation: Department of Pharmacy, University of Salerno, Via Ponte don Melillo 1, 84084 Fisciano, Salerno, Italy.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: De Feo
authors.affiliation: Department of Pharmacy, University of Naples Federico II, Via Domenico Montesano 49, 80131 Naples, Italy.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Caiazzo
authors.affiliation: Department of Pharmacy, University of Salerno, Via Ponte don Melillo 1, 84084 Fisciano, Salerno, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: De Martino
authors.affiliation: Department of Pharmacy, University of Naples Federico II, Via Domenico Montesano 49, 80131 Naples, Italy.
authors.forename: Carla
authors.initials: C
authors.lastname: Cicala
authors.affiliation: Department of Pharmacy, University of Naples Federico II, Via Domenico Montesano 49, 80131 Naples, Italy.
authors.forename: Nicola
authors.initials: N
authors.lastname: Mascolo
mesh_terms: D045784:Abietanes / Q000494:pharmacology*; D000107:Acetylation; D000818:Animals; D001760:Bleeding Time; D001792:Blood Platelets / Q000187:drug effects* / Q000201:enzymology; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D006487:Hemostasis / Q000187:drug effects; D008297:Male; D051379:Mice; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D010766:Phosphorylation; D008517:Phytotherapy; D010936:Plant Extracts / Q000302:isolation & purification / Q000494:pharmacology*; D018517:Plant Roots; D010946:Plants, Medicinal; D010974:Platelet Aggregation / Q000187:drug effects*; D010975:Platelet Aggregation Inhibitors / Q000302:isolation & purification / Q000494:pharmacology*; D010979:Platelet Function Tests; D017208:Rats, Wistar; D027561:Salvia miltiorrhiza / Q000737:chemistry; D015398:Signal Transduction / Q000187:drug effects*; D013997:Time Factors; D014404:Tubulin / Q000378:metabolism
chemical_list: D045784:Abietanes; D010936:Plant Extracts; D010975:Platelet Aggregation Inhibitors; D014404:Tubulin; C021751:tanshinone; C535149:Mapk1 protein, rat; D019950:Mitogen-Activated Protein Kinase 1
keywords: Adenosine diphosphate (PubChem CID: 6022); Antiplatelet; Aspirin (PubChem CID: 2244); Cardiovascular; Collagen (PubChem CID: 6913668); Diterpenoids; Tanshinone IIA; Tanshinone IIA (PubChem CID: 164676); Traditional chinese medicine