RESEARCH ARTICLE Proteomic analysis of the pathophysiological process involved in the antisnake venom effect of Mucuna pruriens extract Roberto Guerranti 1 , Ifeanyi G. Ogueli 1 , Erica Bertocci 1 , Chiara Muzzi 1 , John C. Aguiyi 2 , Riccardo Cianti 3 , Alessandro Armini 3 , Luca Bini 3 , Roberto Leoncini 1 , Enrico Marinello 1 and Roberto Pagani 1 1 Department of Internal Medicine, Endocrine-Metabolic Sciences and Biochemistry, University of Siena, Siena, Italy 2 Department of Pharmacology and Clinical Pharmacy, University of Jos, Jos, Nigeria 3 Department of Molecular Biology, University of Siena, Siena, Italy Previously, we reported the antisnake venom properties of a Mucuna pruriens seed extract (MPE) and tested its in vivo efficacy against Echis carinatus venom (EV) in short- (1 injection) and long- term (three weekly injections) treatments. The aim of the present study was to investigate plasma proteome changes associated with MPE treatments and identify proteins responsible for survival of envenomated mice (CHALLENGED mice). Six treatment groups were studied. Three control groups: one saline, one short-term and one long-term MPE treatment. One group received EV alone. Two test groups received EV with either a short-term or long-term MPE treatment (CHALLENGED mice). The plasma from each group was analysed by 2-DE/MALDI-TOF MS. The most significant changes with treatment were: albumin, haptoglobin, fibrinogen, serum amyloid A and serum amyloid P. Most of these changes were explained by EV effects on coagu- lation, inflammation and haemolysis. However, MPE treatments prevented the EV-induced ele- vation in HPT. Consequently, HPT levels were similar to controls in the plasma of CHAL- LENGED mice. The plasma of CHALLENGED mice showed substantial proteomic modifica- tions. This suggests the mechanism of MPE protection involves the activation of counterbalancing processes to compensate for the imbalances caused by EV. Received: March 16, 2007 Revised: September 27, 2007 Accepted: October 15, 2007 Keywords: 2-DE / Mouse plasma proteome / Mucuna pruriens / Plant extract / Snake venom 402 Proteomics 2008, 8, 402–412 1 Introduction The well-known high mortality and morbidity rate of snake- bite victims, especially in tropical and subtropical countries, is estimated to be about 100 000 per year [1]. The traditional treatment for envenomation is the administration of specific antivenoms that are usually developed in animals, primarily horses. However, problems with this approach include the relative unavailability of specific antivenoms and the varia- bility in venom composition [2]. Recent improvements in antivenom production technology have led to the production of Fab fragments [3]. Nevertheless, the efficacy of this remedy is dependent on its immediate administration, which is lim- Correspondence: Dr. Roberto Guerranti, Department of Internal Medicine, Endocrine-Metabolic Sciences and Biochemistry, Uni- versity of Siena, Via A. Moro 2, 53100 Siena, Italy E-mail: guerranti@unisi.it Fax: 139-0577234285 Abbreviations: APP, acute phase proteins; CHALLENGED, MPE treated mice that survived envenomation; EV, Echis carinatus venom; FIBB, FIBG, b and g chains of fibrinogen; HPT, haptoglo- bin; MPE, Mucuna pruriens extract; SAA, serum amyloid A; SAP, serum amyloid P DOI 10.1002/pmic.200700265  2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.proteomics-journal.com