Effects of cocktail of four local Malaysian medicinal plants (Phyllanthus spp.) against dengue virus 2

BMC Complementary and Alternative Medicine

Shamala Devi Sekaran²  Indu Bala Jaganath¹  Bobby Joe Payne⁶  Rishya Manikam³  Kien Chai Ong⁵  Seok Mui Wang⁴  Anusyah Rathkrishnan²  Yin Quan Tang²  Sau Har Lee² 

[1] Biotechnology Centre, Malaysia Agricultural Research and Development Institute (MARDI), Serdang, Malaysia;Department of Medical Microbiology, Faculty of Medicine, University Malaya, Kuala Lumpur, Malaysia;Department of Trauma & Emergency Medicine, UMMC, Kuala Lumpur, Malaysia;Institute for Medical Molecular Biotechnology, University of Islamic Technology Malaysia, Kuala Lumpur, Malaysia;Department of Molecular Medicine, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia;American College of Veterinary Pathology, Madison, USA

关键词: Antiviral;    Dengue;    Phyllanthus;   

Others  :  1221054
DOI  :  10.1186/1472-6882-13-192

 received in 2013-03-07, accepted in 2013-07-24,  发布年份 2013

【 摘 要 】

Background

The absence of commercialized vaccines and antiviral agents against dengue has made the disease a major health concern around the world. With the current dengue virus transmission rate and incidences, the development of antiviral drugs is of vital need. The aim of this project was to evaluate the possibility of developing a local medicinal plant, Phyllanthus as an anti-dengue agent.

Methods

Cocktail (aqueous and methanolic) extracts were prepared from four species of Phyllanthus (P.amarus, P.niruri, P.urinaria, and P.watsonii) and their polyphenolic compounds were identified via HPLC and LC-MS/MS analysis. MTS assay was then carried out to determine the maximal non-toxic dose (MNTD) of the extracts, followed by screening of the in vitro antiviral activity of aqueous cocktail extracts against DENV2 by means of time-of-addition (pre-, simultaneous and post-) using RT-qPCR. The differentially expressed proteins in the treated and infected cells were analysed with two dimensional gel electrophoresis experiments.

Results

Several active compounds including gallic acid, geraniin, syringin, and corilagen have been identified. The MNTD of both aqueous and methanolic extracts on Vero cells were 250.0 μg/ml and 15.63 μg/ml respectively. Phyllanthus showed strongest inhibitory activity against DENV2 with more than 90% of virus reduction in simultaneous treatment. Two-dimensional analysis revealed significantly altered levels of thirteen proteins, which were successfully identified by tandem MS (MS/MS). These altered proteins were involved in several biological processes, including viral entry, viral transcription and translation regulations, cytoskeletal assembly, and cellular metabolisms.

Conclusions

Phyllanthus could be potentially developed as an anti-DENV agent.

【 授权许可】

   
2013 Lee et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Beatty ME, Stone A, Fitzsimons DW, Hanna JN, Lam SK, Vong S, Guzman MG, Mendez-Galvan JF, Halstead SB, Letson GW, et al.: Best practices in dengue surveillance: a report from the Asia-Pacific and Americas Dengue Prevention Boards. PLoS Negl Trop Dis 2010, 4(11):e890.
• [2]Murrell S, Wu SC, Butler M: Review of dengue virus and the development of a vaccine. Biotechnol Adv 2011, 29(2):239-247.
• [3]Noble CG, Chen YL, Dong H, Gu F, Lim SP, Schul W, Wang QY, Shi PY: Strategies for development of Dengue virus inhibitors. Antiviral Res 2010, 85(3):450-462.
• [4]Steuer C, Gege C, Fischl W, Heinonen KH, Bartenschlager R, Klein CD: Synthesis and biological evaluation of alpha-ketoamides as inhibitors of the Dengue virus protease with antiviral activity in cell-culture. Bioorg Med Chem 2011, 19(13):4067-4074.
• [5]Gutsche I, Coulibaly F, Voss JE, Salmon J, d’Alayer J, Ermonval M, Larquet E, Charneau P, Krey T, Megret F, et al.: Secreted dengue virus nonstructural protein NS1 is an atypical barrel-shaped high-density lipoprotein. Proc Natl Acad Sci U S A 2011, 108(19):8003-8008.
• [6]TDR/WHO: Dengue: guidelines for diagnosis, treatment, prevention and control. Geneva: World Health Organization; 2009.
• [7]Halstead SB, Porterfield JS, O’Rourke EJ: Enhancement of dengue virus infection in monocytes by flavivirus antisera. AmJTrop Med Hyg 1980, 29(4):638-642.
• [8]Appanna R, Huat TL, See LL, Tan PL, Vadivelu J, Devi S: Cross-reactive T-cell responses to the nonstructural regions of dengue viruses among dengue fever and dengue hemorrhagic fever patients in Malaysia. Clin Vaccine Immunol 2007, 14(8):969-977.
• [9]Chang J, Schul W, Yip A, Xu X, Guo JT, Block TM: Competitive inhibitor of cellular alpha-glucosidases protects mice from lethal dengue virus infection. Antiviral Res 2011, 92(2):369-371.
• [10]Kato D, Era S, Watanabe I, Arihara M, Sugiura N, Kimata K, Suzuki Y, Morita K, Hidari KI, Suzuki T: Antiviral activity of chondroitin sulphate E targeting dengue virus envelope protein. Antiviral Res 2010, 88(2):236-243.
• [11]Jacobs M: Dengue. Medicine 2005, 33(7):18-20.
• [12]Piraino F, Brandt CR: Isolation and partial characterization of an antiviral, RC-183, from the edible mushroom Rozites caperata. Antiviral Res 1999, 43(2):67-78.
• [13]Herrmann EC Jr, Kucera LS: Antiviral substances in plants of the mint family (labiatae). 3. Peppermint (Mentha piperita) and other mint plants. Proc Soc Exp Biol Med 1967, 124(3):874-878.
• [14]Aswal BS, Goel AK, Kulshrestha DK, Mehrotra BN, Patnaik GK: Screening of Indian plants for biological activity: Part XV. Indian J Exp Biol 1996, 34(5):444-467.
• [15]Lee-Huang S, Zhang L, Huang PL, Chang YT: Anti-HIV activity of olive leaf extract (OLE) and modulation of host cell gene expression by HIV-1 infection and OLE treatment. Biochem Biophys Res Commun 2003, 307(4):1029-1037.
• [16]Yamada K, Ogawa H, Hara A, Yoshida Y, Yonezawa Y, Karibe K, Nghia VB, Yoshimura H, Yamamoto Y, Yamada M, et al.: Mechanism of the antiviral effect of hydroxytyrosol on influenza virus appears to involve morphological change of the virus. Antiviral Res 2009, 83(1):35-44.
• [17]Zhu W, Chiu LC, Ooi VE, Chan PK, Ang PO Jr: Antiviral property and mechanisms of a sulphated polysaccharide from the brown alga Sargassum patens against Herpes simplex virus type 1. Phytomedicine 2006, 13(9–10):695-701.
• [18]Cui X, Wang Y, Kokudo N, Fang D, Tang W: Traditional Chinese medicine and related active compounds against hepatitis B virus infection. Biosci Trends 2010, 4(2):39-47.
• [19]Cacciotti P, Libener R, Betta P, Martini F, Porta C, Procopio A, Strizzi L, Penengo L, Tognon M, Mutti L, et al.: SV40 replication in human mesothelial cells induces HGF/Met receptor activation: a model for viral-related carcinogenesis of human malignant mesothelioma. Proc Natl Acad Sci U S A 2001, 98(21):12032-12037.
• [20]Eldeen I, Seow E, Abdullah R, Sulaiman S: In vitro antibacterial, antioxidant, total phenolic contents and anti-HIV-1 reverse transcriptase activities of extracts of seven Phyllanthus sp. S Afr J Bot 2010, 77(1):75-79.
• [21]Schmelzer GH: Plant resources of tropical Africa: medicinal plants. Prota 2008, 11:270-271.
• [22]Tang YQ, Jaganath IB, Sekaran SD: Phyllanthus spp. induces selective growth inhibition of PC-3 and MeWo human cancer cells through modulation of cell cycle and induction of apoptosis. PLoS One 2010, 5(9):e12644.
• [23]Lee SH, Jaganath IB, Wang SM, Sekaran SD: Antimetastatic effects of Phyllanthus on human lung (A549) and breast (MCF-7) cancer cell lines. PLoS One 2011, 6(6):e20994.
• [24]Yong YK, Thayan R, Chong HT, Tan CT, Sekaran SD: Rapid detection and serotyping of dengue virus by multiplex RT-PCR and real-time SYBR green RT-PCR. Singapore Med J 2007, 48(7):662-668.
• [25]Venkateswaran PS, Millman I, Blumberg BS: Effects of an extract from Phyllanthus niruri on hepatitis B and woodchuck hepatitis viruses: in vitro and in vivo studies. Proc Natl Acad Sci U S A 1987, 84(1):274-278.
• [26]Zhou S, Xu C, Zhou N, Huang Y, Huang L, Chen X, Hu Y, Liao Y: Mechanism of protective action of Phyllanthus urinaria L. against injuries of liver cells]. Zhongguo Zhong yao za zhi = Zhongguo zhongyao zazhi = China journal of Chinese materia medica 1997, 22(2):109.
• [27]Rajeshkumar NV, Kuttan R: Phyllanthus amarus extract administration increases the life span of rats with hepatocellular carcinoma. J Ethnopharmacol 2000, 73(1–2):215-219.
• [28]Ott M, Thyagarajan SP, Gupta S: Phyllanthus amarus suppresses hepatitis B virus by interrupting interactions between HBV enhancer I and cellular transcription factors. Eur J Clin Invest 1997, 27(11):908-915.
• [29]Mazumder A, Mahato A, Mazumder R: Antimicrobial potentiality of Phyllanthus amarus against drug resistant pathogens. Nat Prod Res 2006, 20(4):323-326.
• [30]Kloucek P, Polesny Z, Svobodova B, Vlkova E, Kokoska L: Antibacterial screening of some Peruvian medicinal plants used in Calleria District. J Ethnopharmacol 2005, 99(2):309-312.
• [31]Mazunder UK, Gupta M, Rajeshwar Y: Antihyperglycemic effect and antioxidant potential of Phyllanthus niruri (Euphorbiaceae) in streptozotocin induced diabetic rats. Eur Bull Drug Res 2005, 13(1):15-23.
• [32]Raphael KR, Sabu MC, Kuttan R: Hypoglycemic effect of methanol extract of Phyllanthus amarus Schum & Thonn on alloxan induced diabetes mellitus in rats and its relation with antioxidant potential. Indian J Exp Biol 2002, 40(8):905-909.
• [33]Yang CM, Cheng HY, Lin TC, Chiang LC, Lin CC: The in vitro activity of geraniin and 1,3,4,6-tetra-O-galloyl-beta-D-glucose isolated from Phyllanthus urinaria against herpes simplex virus type 1 and type 2 infection. J Ethnopharmacol 2007, 110(3):555-558.
• [34]Yang Y, Zhang L, Fan X, Qin C, Liu J: Antiviral effect of geraniin on human enterovirus 71 in vitro and in vivo. Bioorg Med Chem Lett 2012, 22(6):2209-2211.
• [35]Kwon H-J, Kim H-H, Yoon S, Ryu Y, Chang J, Cho K-O, Rho M-C, Park S-J, Lee W: In Vitro inhibitory activity of Alpinia katsumadai extracts against influenza virus infection and hemagglutination. Virol J 2010, 7(1):307. BioMed Central Full Text
• [36]Sepulveda CS, Garcia CC, Fascio ML, D’Accorso NB, Docampo Palacios ML, Pellon RF, Damonte EB: Inhibition of Junin virus RNA synthesis by an antiviral acridone derivative. Antiviral Res 2012, 93(1):16-22.
• [37]Alvarez AL, del Barrio G, Kouri V, Martinez PA, Suarez B, Parra F: In vitro anti-herpetic activity of an aqueous extract from the plant Phyllanthus orbicularis. Phytomedicine 2009, 16(10):960-966.
• [38]Jindadamrongwech S, Thepparit C, Smith D: Identification of GRP 78 (BiP) as a liver cell expressed receptor element for dengue virus serotype 2. Arch Virol 2004, 149(5):915-927.
• [39]Chavez-Salinas S, Ceballos-Olvera I, Reyes-del Valle J, Medina F, del Angel RM: Heat shock effect upon dengue virus replication into U937 cells. Virus Res 2008, 138(1–2):111-118.
• [40]Delneste Y, Magistrelli G, Gauchat J-F, Haeuw J-F, Aubry J-P, Nakamura K, Kawakami-Honda N, Goetsch L, Sawamura T, Bonnefoy J-Y, et al.: Involvement of LOX-1 in dendritic cell-mediated antigen cross-presentation. Immunity 2002, 17(3):353-362.
• [41]Suksanpaisan L, Cabrera-Hernandez A, Smith DR: Infection of human primary hepatocytes with dengue virus serotype 2. J Med Virol 2007, 79(3):300-307.
• [42]Ishiki Y, Ohnishi H, Muto Y, Matsumoto K, Nakamura T: Direct evidence that hepatocyte growth factor is a hepatotrophic factor for liver regeneration and has a potent antihepatitis effect in vivo. Hepatology 1992, 16(5):1227-1235.
• [43]Kashiwakura Y, Tamayose K, Iwabuchi K, Hirai Y, Shimada T, Matsumoto K, Nakamura T, Watanabe M, Oshimi K, Daida H: Hepatocyte growth factor receptor is a coreceptor for adeno-associated virus type 2 infection. J Virol 2005, 79(1):609-614.
• [44]Voraphani N, Theamboonlers A, Khongphatthanayothin A, Srisai C, Poovorawan Y: Increased level of hepatocyte growth factor in children with dengue virus infection. Ann Trop Paediatr 2010, 30(3):213-218.
• [45]McNiven MA, Thompson HM: Vesicle formation at the plasma membrane and trans-Golgi network: the same but different. Science 2006, 313(5793):1591-1594.
• [46]El-Bacha T, Menezes MM, Azevedo e Silva MC, Sola-Penna M, Da Poian AT: Mayaro virus infection alters glucose metabolism in cultured cells through activation of the enzyme 6-phosphofructo 1-kinase. Mol Cell Biochem 2004, 266(1–2):191-198.
• [47]Ranji A, Boris-Lawrie K: RNA helicases: emerging roles in viral replication and the host innate response. RNA Biol 2010, 7(6):775-787.
• [48]Yang SH, Liu ML, Tien CF, Chou SJ, Chang RY: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) interaction with 3′ ends of Japanese encephalitis virus RNA and colocalization with the viral NS5 protein. J Biomed Sci 2009, 16:40. BioMed Central Full Text
• [49]Nisole S, Stoye JP, Saib A: TRIM family proteins: retroviral restriction and antiviral defence. Nat Rev Microbiol 2005, 3(10):799-808.
• [50]Yap MW, Nisole S, Lynch C, Stoye JP: Trim5α protein restricts both HIV-1 and murine leukemia virus. Proc Natl Acad Sci U S A 2004, 101(29):10786.
• [51]Nitsche C, Behnam MA, Steuer C, Klein CD: Retro peptide-hybrids as selective inhibitors of the Dengue virus NS2B-NS3 protease. Antiviral Res 2012, 94(1):72-79.
• [52]Limjindaporn T, Wongwiwat W, Noisakran S, Srisawat C, Netsawang J, Puttikhunt C, Kasinrerk W, Avirutnan P, Thiemmeca S, Sriburi R, et al.: Interaction of dengue virus envelope protein with endoplasmic reticulum-resident chaperones facilitates dengue virus production. Biochem Biophys Res Comm 2009, 379(2):196-200.
• [53]Daikoku T, Kudoh A, Sugaya Y, Iwahori S, Shirata N, Isomura H, Tsurumi T: Postreplicative mismatch repair factors Are recruited to Epstein-Barr virus replication compartments. J Biol Chem 2006, 281(16):11422-11430.
• [54]Mohni KN, Mastrocola AS, Bai P, Weller SK, Heinen CD: DNA mismatch repair proteins Are required for efficient herpes simplex virus 1 replication. J Virol 2011, 85(23):12241-12253.
• [55]Kanlaya R, Pattanakitsakul S-n, Sinchaikul S, Chen S-T, Thongboonkerd V: Vimentin interacts with heterogeneous nuclear ribonucleoproteins and dengue nonstructural protein 1 and is important for viral replication and release. Mol BioSys 2010, 6(5):795-806.