Antituberculosis Activity of Brotowali (Tinospora crispa) Extract and Fractions against Mycobacterium tuberculosis using Microplate Alamar Blue Assay Method

Retno Wahyuningrum(1*), Ritmaleni Ritmaleni(2), Tatang Irianti(3), Subagus Wahyuono(4), Takushi Kaneko(5), Titik Nuryastuti(6)

(1) Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta, Indonesia Faculty of Pharmacy Universitas Muhammadiyah Purwokerto, Indonesia
(2) Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta
(3) Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta
(4) Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta
(5) Tuberculosis Alliance, New York
(6) Faculty of Medicine Universitas Gadjah Mada, Yogyakarta
(*) Corresponding Author


Tuberculosis (TB), in which caused by pathogenic bacteria, Mycobacterium tuberculosis, has become the major causes of death among all of infectious diseases. The increasing incidence of multidrug-resistant tuberculosis (MDR-TB) and extensively drug-resistant tuberculosis (XDR-TB) has created a need to discover a new antituberculosis drug candidate. The aim of this study was to screen extract and fractions of Tinospora crispa for activity against Mycobacterium tuberculosis H37Rv using the Microplate Alamar Blue Assay (MABA) method. T. crispa extract was prepared by maceration in ethanol (96%) and antituberculosis activity was carried out using MABA method. The result of this study showed that ethanolic extract of T. crispa exhibit antituberculosis activity with minimum inhibition concentration of 12.5 mg/ml.

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Abu, M.N., Samat, S., Kamarapani, N., Nor Hussein, F., Wan Ismail, W.I., Hassan, H.F., 2015. Tinospora crispa Ameliorates Insulin Resistance Induced by High Fat Diet in Wistar Rats. Evid. Based Complement. Alternat. Med. 2015, 1–6.

Alvin, A., Miller, K.I., Neilan, B.A., 2014. Exploring the potential of endophytes from medicinal plants as sources of antimycobacterial compounds. Microbiol. Res. 169, 483–495.

Borges-Argáez, R., Canche-Chay, C.I., Peña-Rodríguez, L.M., Said-Fernández, S., Molina-Salinas, G.M., 2007. Antimicrobial activity of Diospyros anisandra. Fitoterapia 78, 370–372.

Camacho-Corona, M. del R., Ramírez-Cabrera, M.A., Santiago, O.G.-, Garza-González, E., Palacios, I. de P., Luna-Herrera, J., 2008. Activity against drug resistant-tuberculosis strains of plants used in Mexican traditional medicine to treat tuberculosis and other respiratory diseases. Phytother. Res. 22, 82–85.

Chang, C.-C., Ho, S.L., Lee, S.-S., 2015. Acylated glucosylflavones as α-glucosidase inhibitors from Tinospora crispa leaf. Bioorg. Med. Chem. 23, 3388–3396.

Chinsembu, K.C., 2016. Tuberculosis and nature’s pharmacy of putative anti-tuberculosis agents. Acta Trop. 153, 46–56.

Collins, L., Franzblau, S.G., 1997. Microplate alamar blue assay versus BACTEC 460 system for high-throughput screening of compounds against Mycobacterium tuberculosis and Mycobacterium avium. Antimicrob. Agents Chemother. 41, 1004–1009.

Elkington, B.G., Sydara, K., Newsome, A., Hwang, C.H., Lankin, D.C., Simmler, C., Napolitano, J.G., Ree, R., Graham, J.G., Gyllenhaal, C., Bouamanivong, S., Souliya, O., Pauli, G.F., Franzblau, S.G., Soejarto, D.D., 2014. New finding of an anti-TB compound in the genus Marsypopetalum (Annonaceae) from a traditional herbal remedy of Laos. J. Ethnopharmacol. 151, 903–911.

Franzblau, S.G., DeGroote, M.A., Cho, S.H., Andries, K., Nuermberger, E., Orme, I.M., Mdluli, K., Angulo-Barturen, I., Dick, T., Dartois, V., Lenaerts, A.J., 2012. Comprehensive analysis of methods used for the evaluation of compounds against Mycobacterium tuberculosis. Tuberculosis 92, 453–488.

Franzblau, S.G., Witzig, R.S., Mclaughlin, J.C., Torres, P., Madico, G., Hernandez, A., Degnan, M.T., Cook, M.B., Quenzer, V.K., Ferguson, R.M., Gilman, R.H., 1998. Rapid, low-technology MIC determination with clinical Mycobacterium tuberculosis isolates by using the microplate Alamar Blue assay. J. Clin. Microbiol. 36, 362–366.

García, A., Bocanegra-García, V., Palma-Nicolás, J.P., Rivera, G., 2012. Recent advances in antitubercular natural products. Eur. J. Med. Chem. 49, 1–23.

Gautam, R., Saklani, A., Jachak, S.M., 2007. Indian medicinal plants as a source of antimycobacterialagents. J. Ethnopharmacol. 110, 200–234.

Kementerian Kesehatan Republik Indonesia, 2012, Petunjuk Teknis Pemeriksaan Biakan, Identifikasi, Dan Uji Kepekaan Mycobacterium tuberculosis pada Media Padat, Jakarta

Keshavjee, S., Farmer, P.E., 2012. Tuberculosis, Drug Resistance, and the History of Modern Medicine. N. Engl. J. Med. 367, 931–936.

Koay, Y., Koay, F., 2013. A Review of the Secondary Metabolites and Biological Activities of <i>Tinospora crispa</i> (Menispermaceae). Trop. J. Pharm. Res., 12, 641-649

Kuete, V., 2010. Potential of Cameroonian Plants and Derived Products against Microbial Infections: A Review. Planta Med. 76, 1479–1491.

Mohamad, S., Zin, N.M., Wahab, H.A., Ibrahim, P., Sulaiman, S.F., Zahariluddin, A.S.M., Noor, S.S.M., 2011. Antituberculosis potential of some ethnobotanically selected Malaysian plants. J. Ethnopharmacol. 133, 1021–1026.

Nguta, J.M., Appiah-Opong, R., Nyarko, A.K., Yeboah-Manu, D., Addo, P.G.A., 2015. Current perspectives in drug discovery against tuberculosis from natural products. Int. J. Mycobacteriology 4, 165–183.

Okunade, A.L., Elvin-Lewis, M.P.F., Lewis, W.H., 2004. Natural antimycobacterial metabolites: current status. Phytochemistry 65, 1017–1032.

Prabu, A., Hassan, S., Prabuseenivasan, Shainaba, A.S., Hanna, L.E., Kumar, V., 2015. Andrographolide: A potent antituberculosis compound that targets Aminoglycoside 2′-N-acetyltransferase in Mycobacterium tuberculosis. J. Mol. Graph. Model. 61, 133–140.

Tekwu, E.M., Askun, T., Kuete, V., Nkengfack, A.E., Nyasse, B., Etoa, F.-X., Beng, V.P., 2012. Antibacterial activity of selected Cameroonian dietary spices ethno-medically used against strains of Mycobacterium tuberculosis. J. Ethnopharmacol. 142, 374–382.

Tosun, F., Kızılay, Ç.A., Şener, B., Vural, M., Palittapongarnpim, P., 2004. Antimycobacterial screening of some Turkish plants. J. Ethnopharmacol. 95, 273–275.

World Health Organization, 2015. Global tuberculosis report 2015. World Health Organization, [S.l.].


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