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Mixture of Uncaria and Tabebuia Extracts Are Mixture of Uncaria and Tabebuia Extracts are Potentially Chemopreventive in CBA/Ca Mice - A Long-term Experiment Ferenc Budán, István Szabo, Timea Varjas, Ghodratollah Nowrasteh, Tamás Dávid, Péter Gergely, Zsuzsa Varga, Kornélia Molnár, Balázs Kádár, Zsuzsa Orsos, et al. To cite this version: Ferenc Budán, István Szabo, Timea Varjas, Ghodratollah Nowrasteh, Tamás Dávid, et al.. Mixture of Uncaria and Tabebuia Extracts are Potentially Chemopreventive in CBA/Ca Mice - A Long-term Experiment. Phytotherapy Research, Wiley, 2010, 25 (4), pp.493. 10.1002/ptr.3281. hal-00599836 HAL Id: hal-00599836 https://hal.archives-ouvertes.fr/hal-00599836 Submitted on 11 Jun 2011 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Phytotherapy Research Mixture of Uncaria and Tabebuia Extracts are Potentially Chemopreventive in CBA/Ca Mice - A Long-term Experiment Journal: Phytotherapy Research ManuscriptFor ID: PTR-10-0137.R1 Peer Review Wiley - Manuscript type: Full Paper Date Submitted by the 07-Jun-2010 Author: Complete List of Authors: Budán, Ferenc; University of Pécs, Faculty of Medicine, Institute of Public Health Szabo, István; University of Pecs, Faculty of Medicine, Institute of Public Health Varjas, Timea; University of Pécs, Faculty of Medicine, Institut of Public Health Nowrasteh, Ghodratollah; University of Pécs, Faculty of Medicine, Institute of Public Health Dávid, Tamás; CoD Cancer Information & Research Foundation Gergely, Péter; University of Debrecen, Institute of Forensic Medicine Varga, Zsuzsa; University of Pecs, Faculty of Medicine Molnár, Kornélia; Office of the Chief Medical Officer Kádár, Balázs; University of Pecs, Faculty of Medicine Orsos, Zsuzsa; University of Pécs, Faculty of Medicine, Institute of Public Health Kiss, István; University of Pecs, Faculty of Medicine, Institute of Public Health Ember, István; University of Pecs, Faculty of Medicine, Institute of Public Health Gene expression, carcinogenesis, chemoprevention, DMBA, Uncaria, Keyword: Tabebuia http://mc.manuscriptcentral.com/ptr Page 1 of 21 Phytotherapy Research 1 2 3 Mixture of Uncaria and Tabebuia Extracts are Potentially Chemopreventive in CBA/Ca 4 5 Mice - A Long-term Experiment 6 1 1 1 7 FERENC BUDÁN , ISTVÁN SZABÓ , TÍMEA VARJAS , GHODRATOLLAH 8 NOWRASTEH 1, TAMÁS DÁVID 2, PÉTER GERGELY 3, ZSUZSA VARGA 4, KORNÉLIA 9 MOLNÁR 5, BALÁZS KÁDÁR 1, ZSUZSA ORSÓS 1, ISTVÁN KISS 1 and ISTVÁN EMBER 1 10 11 1 Institute of Public Health, Medical School, University of Pécs, Szigeti str. 12, H-7643 Pécs; 12 2 13 CoD Cancer Information & Research Foundation, Vörösmarty str. 15, H- 9155 Lébény; 14 3Institute of Forensic Medicine, University of Debrecen, Nagyerdei krt. 98., H-4012, 15 Debrecen; 16 4Country Hospital of Baranya, Department of Oncology, H-7623, Pécs; 17 5 18 Office of the Chief Medical Officer, Gyáli str. 2-6, H-1097, Budapest. 19 20 For Peer Review 21 22 23 Correspondence to: Ferenc Budán 24 25 [email protected] 26 Address: Szigeti str. 12, H-7624 Pécs, Hungary 27 Phone: +36-72-536-394 28 Fax: +36-72-536-395 29 30 31 Experimental work 32 33 34 Budán et al: Uncaria and Tabebuia are Chemopreventive in Long-term Mice Experiment 35 36 37 38 Key Words: Gene expression, carcinogenesis, chemoprevention, DMBA, Uncaria, 39 Tabebuia 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 http://mc.manuscriptcentral.com/ptr Phytotherapy Research Page 2 of 21 1 2 3 Abstract. 4 5 6 A long-term experimental animal model was developed by our research group for the 7 8 evaluation of potential chemopreventive effects. The inhibitory effects of agents on 9 10 carcinogen (7,12-Dimetilbenz[a]anthracene ( DMBA)) induced molecular epidemiological 11 12 13 biomarkers, in this case the expression of key onco/suppressor genes were investigated. 14 15 Expression pattern of c-myc , Ha-ras , Bcl-2, K-ras protoonco and p53 tumoursuppressor genes 16 17 18 were studied to elucidate early carcinogenic and potential chemopreventive effects. 19 TM 20 Consumption of so calledFor "CoD Peer tea” (abbreviation Review of Claw of Dragon tea) containing bark 21 22 of Uncaria guianensis , Cat's Claw (Uncaria sp. U. tomentosa ) and Palmer trumpet-tree 23 24 25 (Tabebuia sp. T. avellanedae ) was able to decrease the DMBA-induced onco/suppressor gene 26 27 overexpression in a short-term animal experiment. In following study CBA/Ca mice were 28 29 treated with 20 mg/bwkg DMBA intraperitoneally (ip. ) and expression patterns of 30 31 32 onco/suppressor genes were examined at several time intervals. According to the examined 33 34 gene expression patterns in this long-term experiment the chemopreventive effect of CoD TM 35 36 tea consumption could be confirmed. 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 2 http://mc.manuscriptcentral.com/ptr Page 3 of 21 Phytotherapy Research 1 2 3 In our previous studies a unique animal test model was developed through the investigation of 4 5 6 onco/suppressor genes expression patterns, as molecular epidemiological biomarkers (Ember 7 8 et al. , 1998 ). 7,12-dimetilbenz[a]anthracene ( DMBA) is a pluripotent and complete 9 10 carcinogen, therefore it is applied to induce carcinomas in animal experiments. DMBA 11 12 13 increases the expression of c-myc , Ha-ras , Bcl-2, K-ras protoonco (Gyöngyi and Somlyai, 14 15 2002) and consequently p53 tumoursuppressor genes in the lungs and in the liver, as well 16 17 18 (Sándor et al. , 1995 ) in several animal species and also in both sex, indicating both 19 20 carcinogenic exposureFor and early stepsPeer of tumour Review formation (Ember et al. , 1998 ). The purpose 21 22 was to study the early biological effect of DMBA and the protective effect of some potential 23 24 TM 25 chemopreventive agents in so called “CoD tea" (abbreviation of Claw of Dragon tea). 26 27 Consumption of “CoD TM tea”, which contains extract of Uncaria guianensis , Cat's Claw 28 29 (Uncaria sp. U. tomentosa ) and Palmer trumpet-tree (Tabebuia sp. T. avellanedae ), was able 30 31 32 to decrease the DMBA-induced overexpression of the genes mentioned above, in a short-term 33 34 animal experiment (Orsós et al. , 2007 ). According to pharmacological studies Uncaria species 35 36 have cytotoxic, anti-inflammatory, antiviral, immunostimulative, antioxidant and antibacterial 37 38 39 properties (Heitzman et al. , 2005 ). Keplinger et al. confirmed that the pentacyclic oxindol 40 41 alkaloides (POA) are the main biological active components of Uncaria species, which in 42 43 44 vitro activate lymphocyte-proliferation regulating factor in endothelial cells (Laus et al. , 45 46 1997; Keplinger et al. , 1999 ). The bark of Palmer trumpet-tree is abundant in lapachol (2- 47 48 hydroxy-3-(3-methyl-2-butenyl)-1,4-naphthoquinone) (2-7%), lapachone and xyloidone (de 49 50 51 Santana et al. , 1968 ), which are anticarcinogenic (Ueda et al. , 1994 ) and inhibit the growth of 52 53 the human keratinocyte cell line HaCaT culture in vitro (Müller et al. , 1999 ). Our research 54 55 group supposed that CoD TM tea consumption was able to decrease the overexpression of 56 57 58 certain key onco/suppressor genes. 59 60 3 http://mc.manuscriptcentral.com/ptr Phytotherapy Research Page 4 of 21 1 2 3 In this long term study the effect of CoD TM tea was investigated on the expression of c-myc , 4 5 6 Ha-ras , Bcl-2, K-ras and p53 onco/suppressor genes in CBA/Ca mice test model (Varga et 7 8 al. , 1991; Ember et al. , 1992; Gyöngyi and Somlyai 2000 ). 9 10 11 12 13 Materials and methods 14 15 Animals and Treatments. 16 17 TM 18 Infuse of CoD tea (D/Eu reg. numb.: PZN Deutschland: 4226037) was prepared according 19 20 to the instruction of theFor manufacturer. Peer Review 21 22 10g of the powder was suspended in 750 ml tap water and left to soak for twelve hours, and 23 24 25 then it was heated to 80 ºC for thirty minutes. After filtration and water refill, the infuse was 26 27 kept in a closed bottle avoiding light at 4 ºC. 28 29 250 ml extract is advised to be consumed three times a day, therefore the human dose is 0.167 30 31 32 g powder/kg per os (calculated for 60 kg mean body weight). 33 34 Calculated equivalent dose for mice is 0.0033 g powder in 3 ml tap water. 35 36 Quantity of total pentacyclic oxindol alkaloides (POA) is supposed to be 0.1-1.12%, based on 37 38 39 the results of Csapi et al. ( Csapi et al. , 2008 ). 40 41 Six- to eight-week-old (20±4 g) conventionally kept CBA/Ca inbred H-2K haplotype mice (6 42 43 44 males and 6 females in each group) were used for this experiment according to the following 45 TM 46 arrangement: control groups, DMBA treated groups, CoD consuming groups and DMBA 47 48 treated groups with CoD TM consumption. These setups were measured at five time intervals 49 50 51 after the first DMBA treatment: 1 week, 1 month, 3, 6 and 12 months later. 52 53 Control groups were given tap water and treated with ip. corn oil.
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