Sains
Malaysiana 50(10)(2021): 3085-3094
http://doi.org/10.17576/jsm-2021-5010-21
Zingiber zerumbet (L.)
Smith Hexane Crude Extract Caused DNA Damage on Leptospira spp.
(Ekstrak
Mentah Heksana Zingiber zerumbet (L.) Smith Menyebabkan Kerosakan DNA
pada Leptospira spp.)
FARAH
WAHIDA IBRAHIM1, NURUL AMIRAH ABDUL AZIZ1, LATIFAH
IBRAHIM2, NURUL FARHANA JUFRI1 & ASMAH HAMID1*
1Center for
Toxicology and Health Risk Studies (CORE), Faculty of Health Sciences, Universiti
Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Federal
Territory, Malaysia
2Biotechnology Unit,
Institute for Medical Research, Jalan Pahang, 50588 Kuala Lumpur, Federal
Territory, Malaysia
Diserahkan: 25 Mac 2020/Diterima: 18 Februari 2021
ABSTRACT
Numerous attempts have been made to
control leptospirosis by using chemoprophylaxis, but with limited success. The
present study was done to investigate the antileptospiral potential of hexane,
ethyl acetate and methanol extracts of Zingiber zerumbet rhizomes. The extracts were assayed for
antileptospiral activity using broth microdilution method against Leptospira interrogans (serovar Batavie, Canicola, Australis) and Leptospira biflexa serovar
Patoc. The Z. zerumbet hexane extract exhibited antileptospiral
activity, with IC50 values of 248 μg/mL against L.
interrogans serovar Canicola, IC50 of 125 μg/mL against L.
interrogans serovar Australis, IC50 of 15.63 μg/mL against L.
interrogans serovar Batavie and IC50 of 109 μg/mL against L.
biflexa serovar Patoc. However, both ethyl acetate and methanol extracts did
not show any distinct antileptospiral activity. Since the hexane extract of Z.
zerumbet showed antileptospiral activity, the DNA-damaging properties of
this extract were tested according to their IC50 and IC25 values that were specific to each serovars. The DNA-damaging properties were
determined by treating the selected Leptospira spp. with the hexane
extract and subjecting its DNA to electrophoresis and analysis on agarose gels.
The results demonstrated that the hexane extract had DNA-damaging properties
towards L. biflexa serovar Patoc and L. interrogans serovar
Australis, as proven by the appearance of fragmented DNA on the gels. We
conclude that the Z. zerumbet hexane extract could inhibit the growth of Leptospira spp. serovar Patoc and Australis through DNA-damaging activity
and thus, could be a potential antileptospiral agent. Further studies are
needed to investigate the potential of this hexane extract as an
antileptospiral agent using in vivo rat models of leptospirosis.
Keywords:
Antileptospiral; DNA damage; growth inhibition; hexane crude extract; Zingiber zerumbet
ABSTRAK
Banyak usaha telah
dibuat untuk mengawal leptospirosis dengan menggunakan kemoprofilaksis namun kejayaannya
terhad. Penyelidikan ini dijalankan untuk mengkaji potensi antileptospira bagi
ekstrak kasar heksana, etil asetat dan metanol rizom Zingiber zerumbet. Ekstrak telah diuji untuk aktiviti antileptospira menggunakan kaedah
mikropencairan kaldu ke atas Leptospira interogans (serovar Batavie,
Canicola, Australis) dan Leptospira biflexa serovar Patoc.
Ekstrak heksana Z. zerumbet menunjukkan aktivti antileptospira dengan
nilai IC50 pada 248 μg/mL terhadap L. interrogans serovar
Canicola, IC50 pada 125 μg/mL terhadap L. interrogans serovar
Australis, IC50 pada 15.63 μg/mL terhadap L. interrogans serovar Batavie dan IC50 pada 109 μg/mL untuk L. biflexa serovar Patoc. Walau bagaimanapun, kedua-dua ekstrak etil asetat dan
metanol tidak menunjukkan sebarang aktiviti antileptospira yang ketara. Oleh
kerana ekstrak heksana Z. zerumbet menunjukkan aktiviti antileptospira,
keupayaan merosakkan DNA bagi ekstrak ini diuji mengikut nilai IC50 dan IC25 khusus yang diperoleh untuk setiap serovar. Hasil kajian
menunjukkan bahawa ekstrak heksana mempunyai keupayaan untuk merosakkan DNA L. biflexa serovar Patoc dan L. interrogans serovar Australis seperti
yang terbukti dengan kehadiran DNA yang berfragmen pada gel. Sebagai
kesimpulan, ekstrak heksana Z. zerumbet boleh menghalang pertumbuhan Leptospira spp. serovar Patoc dan Australis melalui aktiviti merosakkan DNA dan dengan
itu, berpotensi sebagai agen antileptospira. Kajian lanjut diperlukan untuk
mengkaji potensi ekstrak heksana sebagai agen antileptospira dalam model tikus
leptospirosis secara in vivo.
Kata kunci:
Antileptospira; ekstrak kasar heksana; kerosakan DNA; merencat pertumbuhan; Zingiber zerumbet
RUJUKAN
Abdul,
A., Siddiq, I.A., Adel, S.A., Manal, M.E. & Syam, M.M. 2008. Anti-cancer
and anti-microbial activities of zerumbone from the rhizomes of Zingiber
zerumbet. Int. J. Pharm. 4(4):
301-304.
Abdul Hamid, Z., Budin, S.B., Ng, W.J., Hamid, A.,
Husain, K. & Mohamed, J. 2012. Nephroprotective effects of Zingiber zerumbet Smith ethyl acetate extract against
paracetamol-induced nephrotoxicity and oxidative stress in rats. J. Zhejiang Univ. Sci. B 13(3): 176-185.
Adler, B.
& de la Pena Moctezuma, A. 2010. Leptospira and
leptospirosis. Vet. Microbiol. 140(3-4): 287-296.
Arzouni, J.P., Parola, P., La Scola, B., Postic, D.,
Brouqui, P. & Raoult, D. 2002. Human infection caused by Leptospira
fainei. Emerg. Infect. 2002:
865-868.
Bahaman, A.R. & Ibrahim, A.L. 1988. A review of
leptospirosis in Malaysia. Vet. Res.
Commun. 12(2): 179-189.
Benacer, D., Siti, N.M.Z., Zhu Sim, S., Nizam, M.K.M.K., Galloway, R.L.,
Souris, M. & Thong, K.L. 2016. Determination of Leptospira
borgpetersenii serovar Javanica and Leptospira interrogans serovar
Bataviae as the persistent Leptospira serovars circulating in the urban
rat populations in Peninsular Malaysia. Parasit Vectors 9: 117.
Benacer, D., Who, P.Y., Mohd
Zain, S.N., Amran, F. & Thong, K.L. 2013. Pathogenic and saprophytic Leptospira species in water and soils from selected urban sites in Peninsular
Malaysia. Microbes. Environ. 28(1): 135-140.
Cushnie,
T.P. & Lamb, A.J. 2005. Antimicrobial activity of flavonoids. Int. J. Antimicrob. Agents 26(5):
343-356.
da
Silva, T.M., Pinheiro, C.D., Orlandi, P.P., Pinheiro, C.C. & Pontes, G.S.
2018. Zerumbone from Zingiber zerumbet (L.) Smith: A potential prophylactic and therapeutic agent against the
cariogenic bacterium Streptococcus mutans. BMC
Complement Altern. Med. 18: 301.
Dive, A.M., Bigaignon, G.
& Reynaert, M. 1987. Adult respiratory distress syndrome in leptospira
icterohaemorrhagiae infection. Intensive Care Med. 13(3): 214.
Echeverría,
J., Opazo, J., Mendoza, L., Urzúa, A. & Wilkens, M. 2017.
Structure-activity and lipophilicity relationships of selected antibacterial
natural flavones and flavanones of Chilean flora. Molecules 22(4): E608. doi: 10.3390/molecules22040608.
Evangelista,
K.V. & Coburn, J. 2010. Leptospira as an emerging pathogen: A review of its
biology, pathogenesis and host immune responses. Future Microbiol. 5(9):
1413-1425.
Faine, S., Adler, B. &
Ruta, G. 1974. A mechanism of immunity to leptospirosis. Aust.
J. Exp. Biol. Med. Sci. 52(2): 301-310.
Garba, B., Bahaman, A.R.,
Khairani-Bejo, S., Zakaria, Z. & Mutalib, A.R. 2017. Retrospective study of leptospirosis in
Malaysia. EcoHealth 14(2): 389-398.
Guerra, M.A. 2013. Leptospirosis: Public health perspectives. Biologicals 41(5):
295-297.
Hamid,
A., Ibrahim, F.W., Teoh, H.M., Nasrom, M.N., Eusoff, N., Husain, K. & Abdul
Latif, M. 2018. Zingiber zerumbet L. (Smith) extract alleviates
the ethanol-induced brain damage via its antioxidant activity. BMC Complement Altern. Med. 18(1): 101.
Hamid,
A., Budin, S.B., Mohamed, R.A.P., Manaf, N.A., Yuhana, N.Y., Husain, K., Abd
Hamid, Z. & Mohamed, J. 2011. Role of oxidative stress in the protective
effects of Zingiber zerumbet Smith ethyl-acetate extract against
paracetamol-induced hepatotoxicity in Sprague-Dawley rat. Aust. J. Basic Appl. Sci. 5(8): 1519-1525.
Haque,
M.A., Jantan, I. & Harikrishnan, H. 2018. Zerumbone suppresses the
activation of inflammatory mediators in LPS-stimulated U937 macrophages through
MyD88-dependent NF-κB/MAPK/PI3K-Akt signaling pathways. Int. Immunopharmacol. 55: 312-322.
Husna, M., Nabilah, I. & Wan Nor Amilah, W.A.W. 2018. Anti-microbial activity of aqueous Quercus infectoria gall extract against pathogenic Leptospira. Malays. J. Med. Sci. 25(4): 42-50.
Kader, G., Nikkon, F., Rashid, M.A. & Yeasmin, T.
2011. Antimicrobial activities of the rhizome extract of Zingiber zerumbet Linn. Asian Pac. J. Trop. Biomed. 1(5):
409-412.
Kader, M.G., Habib, M.R., Nikkon, F., Yeasmin, T.,
Rashid, M.A., Rahman, M.M. & Gibbons, S. 2010. Zederone from the rhizomes
of Zingiber zerumbet and its
anti-staphylococcal activity. Boletín
Latinoamericano y del Caribe de Plantas Medicinales y Aromáticas 9(1):
63-68.
Latifah, I., Abdul Halim, A.,
Rahmat, M.S., Faten, N.M., Ubil, Z.E., Asmah, H., Shafariatul Akmar, I.,
Picardeau, M., Siti Haslina, O. & Nasir, M.A. 2017. Isolation by culture
and PCR identification of LipL32 gene of pathogenic Leptospira spp. in
wild rats of Kuala Lumpur. Malays. J. Pathol. 39(2): 161-166.
Matsui,
M., Rouleau, V., Bruyère-Ostells, L. & Goarant, C. 2011. Gene expression
profiles of immune mediators and histopathological findings in animal models of
leptospirosis: Comparison between susceptible hamsters and resistant mice. Infect. Immun. 79: 4480-4492.
Mérien,
F., Amouriaux, P., Perolat, P., Baranton, G. & Saint Girons, I. 1992.
Polymerase chain reaction for detection of Leptospira spp. in clinical
samples. J. Clin. Microbiol. 30(9):
2219-2224.
Mohd Radi, M.F., Hashim, J.H.,
Jaafar, M.H., Hod, R., Ahmad, N., Nawi, M., Baloch, G.M., Ismail, R. &
Farakhin Ayub, N.I. 2018. Leptospirosis outbreak after the 2014 major flooding
event in Kelantan, Malaysia: A spatial-temporal analysis. Am. J. Trop. Med. Hyg. 98(5): 1281-1295.
Muhammad Aklil, A.R., Aznida, M.Z., Azman,
A., Muhammad Haneef, A., Shaifuddin, N.H., Syed Syarizman, S.A.R., Mohammad
Safree, J., Norfazilah, A. & Mohd Rohaizat, H. 2018. Effectiveness of
antibiotic prophylaxis for leptospirosis among adults: A systematic review. Malays. J. Appl. Sci. 3(2): 46-56.
Murray, C.K. & Hospenthal, D.R. 2004. Determination
of susceptibilities of 26 Leptospira sp. serovars to 24 antimicrobial
agents by a broth microdilution technique. Antimicrob. Agents Chemother. 48(10): 4002-4005.
Musa,
A.M., Ibrahim, M.A., Aliyu, A.B., Abdullahi, M.S., Tajuddeen, N., Ibrahim, H.
& Oyewale, A.O. 2015. Chemical composition and antimicrobial activity of
hexane leaf extract of Anisopus mannii
(Asclepiadaceae). J. Intercult.
Ethnopharmacol. 4(2): 129-133.
Nally, J.E., Whitelegge, J.P.,
Bassilian, S., Blanco, D.R. & Lovett, M.A. 2007. Characterization of the
outer membrane proteome of Leptospira interrogans expressed during acute
lethal infection. Infect. Immun. 75(2):
766-773.
Nelson, J., Chairman, K., Ranjit Singh, A.J.A.,
Padmalatha, C. & Hepsibah, B. 2013. Cytomorphological changes and
inhibition of inclusion body formation in Leptospira
Interrogans on treatment with the extracts of Adhatodavasica. Adv. Tech. Biol. Med. 1(1): 1-4.
Paster, B.J., Dewhirst, F.E., Weisburg, W.G.,
Tordoff, L.A., Fraser, G.J., Hespell, R.B., Stanton, T.B., Zablen, L., Mandelco, L. & Woese, C.R. 1991. Phylogenetic analysis of the spirochetes. J. Bacteriol. 173(19): 6101-6109.
Patel, S.S. & Savjani, J.K. 2015. Systematic
review of plant steroids as potential antiinflammatory agents: Current status and
future perspectives. J. Phytopharmacol. 4(2): 121-125.
Patial,
P.K., Sharma, A., Kaur, I. & Cannoo, D.S. 2019. Correlation study among the
extraction techniques, phytochemicals, and antioxidant activity of Nepeta
spicata aerial part. Biocatalysis and Agricultural Biotechnology 20:
101275.
Picardeau, M., Bulach, D.M.,
Bouchier, C., Zuerner, R.L., Zidane, N., Wilson, P.J., Creno, S., Kuczek, E.S.,
Bommezzadri, S., Davis, J.C., McGrath, A., Johnson, M.J., Boursaux-Eude, C.,
Seemann, T., Rouy, Z., Coppel, R.L., Rood, J.I., Lajus, A., Davies, J.K.,
Médigue, C. & Adler, B. 2008. Genome sequence of the
saprophyte Leptospira biflexa provides insights into the evolution of Leptospira and the pathogenesis of leptospirosis. PLoS ONE 3(2): e1607.
Prakash,
V. 2017. Terpenoids as source of anti-inflammatory compounds. Asian J. Pharm. Clin. Res. 10(3): 68-76.
Saengjaruk,
P., Sakolvaree, Y., Maneewatch, S., Tomanakan, K., Tongtawe, P., Tapchaisri, P.
& Chaicumpa, W. 2007. Components of pathogenic Leptospira spp. with potentials for diagnosis of
human leptospirosis. Asian Pac. J. Allergy Immunol. 25(4): 225-232.
Schneider, A.G., Casanovas-Massana, A., Hacker, K.P., Wunder, E.A.J.,
Begon, M., Reis, M.G., Childs, J.E., Costa, F., Lindow, J.C. & Ko, A.I.
2018. Quantification of pathogenic Leptospira in the soils of a
Brazilian urban slum. PLoS Negl. Trop.
Dis. 12(4): e0006415.
Schneider, M.C., Jancloes, M., Buss, D.F.,
Aldighieri, S., Bertherat, E., Najera, P., Galan, D.I., Durski, K. &
Espinal, M.A. 2013. Leptospirosis: A silent epidemic disease. Int. J.
Environ. Res. Public Health 10(12): 7229-7234.
Shafariatul
Akmar, I., Syahirah, S., Fifi Fariza, A., Asmah, H., Latifah, I. & Dayang
Fredalina, B. 2019. In vitro antileptospiral activity of Canarium
odontophyllum Miq. (Dabai) leaves extract. Malays. J. Microb. 15(3): 220-225.
Vinod,
K.K., Lall, C., Vimal, R.R., Vedhagiri, K., Sunish, I.P. & Vijayachari, P.
2018. Can subminimal inhibitory concentrations of antibiotics induce the
formation of biofilm in Leptospira? Microb. Drug Resist. 24(7): 1040-1042.
Voravuthikunchai, S.P., Phongpaichit, S. &
Subhadhirasakul, S. 2005. Evaluation of antibacterial activities of medicinal
plants widely used among aids patients in Thailand. Pharm. Biol. 43(8): 701-706.
Wong,
S.K., Lim, Y.Y. & Chan, E.W.C. 2010. Evaluation of antioxidant,
anti-tyrosinase and antibacterial activities of selected Hibiscus species. Etnobotanical Leaflets 14: 781-796.
Yob,
N.J., Jofrry, S.M., Affandi, M.M.L.K., Salleh, M.Z. & Zakaria, Z.A. 2011. Zingiber zerumbet (L.) Smith: A review
of its ethnomedicinal, chemical, and pharmacological uses. Evid. Based Complement Alternat. Med. 2011: 543216.
Zaki, A.M., Rahim, M.A.A.,
Azme, M.H., Mahmood, N.A., Jeffree, M.S., Ghazi, H.F. & Hassan, M.R. 2018.
Animal reservoirs for Leptospira spp. in South-East Asia: A
meta-analysis. J. Adv. Res. Med. 5(3): 23-31.
Zhang, W., Xie, X., Wu, D., Jin, X., Liu, R., Hu, X., Fu, Y., Ding, Z.,
Zhang, N. & Cao, Y. 2017. Doxycycline attenuates leptospira-induced
il-1β by suppressing nlrp3 inflammasome priming. Front Immunol. 8: 857.
*Pengarang untuk surat-menyurat; email:
asmah0901@ukm.edu.my
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