Activity of chlorpromazine on nfa1 and Mp2CL5 genes of Naegleria fowleri trophozoites

Supathra Tiewcharoen, Jundee Rabablert, Prucksawan Chetannachan, Dusit Worawirunwong, Virach Junnu, Noppamas Pungsub
DOI: 10.4236/health.2011.33032                

Abstract

Amoeba treatment of patients suffering from pri-mary amoebic meningoencephalitis caused by Naegleria fowleri has not been successful. Dam-aged morphology and effect on genes of N. fowleri as the result of its initial interaction with drug may provide clue to the success of treatment. In this study, we investigated the activity of chlorpromazine compared with amphotericin B and voriconazole against N.fowleri Khon Kaen strain using cell based assay and molecular techniques. Scanning electron and light micro-graph showed the drug interaction of treated amoebae with 0.098 ug/ml chlorpromazine was faster than 0.002 ug/ml amphotericin B and 12.5 ug/ml of voriconazole. The morphological cha-racteristics of treated amoebae with Gomori’s trichrome stain correlated to the scanning elec-tron microscope study. The effect of drugs to nfa1 and Mp2CL5 genes of treated amoebae found that at 120 min post exposure, chlorpromazine, voriconazole inhibited both genes except amphotericin B. Most of drug inhibited nfa1 except fluconazole. The results evaluated that chlorpromazine was higher potency and rapidly activity than amphotericin B and voriconazole against N. fowleri trophozoites.

Keywords

Naegleria fowleri; Chlorpromazine; Scanning Micrograph; Gene Expression

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Schuster, F.L. and Visvesvara, G.S. (2004) Free-living amoebae as opportunistic and nonopportunistic patho-gens of humans and animals. International Journal for Parasitology, 34, 1001-1027. doi:10.1016/j.ijpara.2004.06.004
[2]	Ferrante, A. (1981) Comparative sensitivity of Naegleria fowleri to amphotericin B and amphotericin B methyl ester. Transactions of the Royal Society of Tropical Medicine and Hygiene, 76, 476-478. doi:10.1016/0035-9203(82)90141-9
[3]	Soltow, S.M. and Brenner, G.M. (2007) Synergistic ac-tivities of azithromycin and amphotericin B against Nae-gleria fowleri in vitro and in a mouse model of primary amebic meningoencephalitis. Antimicrobial Agents and Chemotherapy, 51, 23-27. doi:10.1016/0035-9203(82)90141-9
[4]	Madison, B.M. (2001) Application of stains in clinical microbiology. Biotechnic & Histochemistry, 76, 119-125. doi:10.1016/j.exppara.2009.03.016
[5]	González-Robles, A., Cristóbal-Ramos, A.R, González- Lázaro, M., Oma?a-Molina, M. and Martínez-Palomo, A. (2009) Naegleria fowleri: Light and electron microscopy study. Experimental Parasitology, 122, 212-217.
[6]	Gálvez, A., Valdivia, E., González-Segura, A., Lebbadi, M., Martínez-Bueno, M. and Maqueda, M. (1993) Puri-fication, characterization, and lytic activity against Nae-gleria fowleri of two amoebicins produced by Bacillus licheniformis A12. Applied and Environmental Microbi-ology, 59, 1480-1486.
[7]	Marciano-Cabral, F. and John, D. T. (1983) Cytopatho-genicity of Naegleria fowleri for rat neuroblastoma cell cultures: scanning electron microscopy study. Infect Im-munity, 40, 1214-1217.
[8]	Tiewcharoen, S., Rabablert, J., Chetanachan, P., Junnu, V., Worawirounwong, D. and Malainual, N. (2008) Scanning electron microscope of Naegleria fowleri Thai strain during cytopathogenesis in human neuroblastoma cells. Parasitology Research, 103, 1119-1123. doi:10.1007/s00436-008-1103-4
[9]	Maclean, R.C., Richardson, D.J., LePardo, R. and Marciano-Cabral, F. (2004) The identification of Naegle-ria fowleri from water and soil samples by nested PCR. Parasitology Research, 93, 211-217. doi:10.1007/s00436-004-1104-x
[10]	Reveiller, F.L., Cabanes, P.A. and Marciano-Cabral, F. (2002) Development of a nested PCR assay to detect the pathogenic free-living amoeba Naegleria fowleri. Para-sitology Research, 88, 443-450. doi:10.1007/s00436-002-0591-x
[11]	Kang, S.Y., Song, K.J., Jeong, S.R., Kim, J.H., Park, S., Kim, K., Kwon, M.H. and Shin, H.J. (2005) Role of the Nfa1 protein in pathogenic Naegleria fowleri cocultured with CHO target cells. Clinical and Diagnostic Labora-tory, 12, 873- 876.
[12]	Reveiller, F. L., Suh, S., Sullivan, K., Cabanes, P.A and Marciano-Cabral, F. (2001) Isolation of a unique mem-brane protein from Naegleria fowleri. Journal of Eu-karyotic Microbiology, 48, 676-682. doi:10.1111/j.1550-7408.2001.tb00208.x
[13]	Madarova, L., Trnkova, K., Feiková, S., Klement, C. and Obernauerová, M. (2010) A real-time PCR diagnostic method for detection of Naegleria fowleri. Experimental Parasitology, 126, 37-41. doi:10.1016/j.exppara.2009.11.001
[14]	Song, K.J., Jeong, S.R., Park, S., Kim, K., Kwon, M.H., Im, K., Pak, J. H. and Shin, H.J. (2006) Naegleria fowleri: Functional expression of the Nfa1 protein in transfected Naegleria gruberi by promoter modification. Experi-mental Parasitology, 112, 115-20. doi:10.1016/j.exppara.2005.10.004
[15]	Tiewcharoen, S., Rabablert, J. and Junnu, V. (2009) In vitro susceptibility of Naegleria fowleri trophozoites to amphotericin B-combined chlorpromazine. Research Journal of Microbiology, 4, 320-333. doi:10.3923/jm.2009.320.333
[16]	Kim, J.H., Jung, S.Y., Lee,Y.J., Song, K.J., Kwon, D., Park, S., Im, K.I. and Shin, H.J. (2008) Effect of thera-peutic chemical agents in vitro and on experimental meningo-encephalitis due to Naegleria fowleri. Antim-icrobial Agents Chemother, 52, 4010-4016. doi:10.1128/AAC.00197-08
[17]	Tiewchreon, S., Malainual, N., Junnu, V., Chetanachan, P. and Rabablert, J. (2008) Cytopathogenesis of Naegleria fowleri Thai strains for cultured human neuroblastoma cells. Parasitology Research, 102, 997-1000. doi:10.1007/s00436-007-0866-3
[18]	Gormori, G. (1950) A rapid one-step trichrome stain American Journal of Clinical Pathology, 20, 661-664.
[19]	Shin, H. J. and Im, K. I. (2004) Pathogenic free-living amoebae in Korea. Korean Journal of Parasitology, 42, 93-119. doi:10.3347/kjp.2004.42.3.93
[20]	Mandell, G. L. and Petri W. A. Jr (1996) Antimicrobial Agents. In: Hardman, J.G. and Limbird, L.E. eds., Good-man and Gilman's—The Pharmacological Basis of Therapeutics. McGraw-Hill, New York, US, 1165-1168.
[21]	Antonios, S. N. (2010) Scanning electron microscopic study of trophozoite and cyst stages of Naegleria fowleri. Journal of the Egyptian Society of Parasitology, 40, 271-276.
[22]	Shin, H.J., Cho, M.S., Jung, S.Y., Kim, H.I, Park, S. and Im, K.I. (2001) Molecular cloning and characterization of a gene encoding a 13?1 kDa antigenic protein of Nae-gleria fowleri. Journal of Eukaryotic Microbiology, 48, 713-717. doi:10.1111/j.1550-7408.2001.tb00211.x