Susceptibility of Toxoplasma gondii to Ethanolic Extract of Tinospora crispa in Vero Cells

Background Toxoplasmosis remains widely distributed globally and is one of the major neglected parasitic zoonotic infections. The infection is still endemic in most parts of the world due to poor control as well as challenges of the currently used medications which can be overcome by using natural products. This study evaluated the effect of ethanolic extract from the stem of Tinospora crispa (EETC) on host cell invasion and intracellular replication of Toxoplasma gondii. Method The stem powder of T. crispa was soaked in absolute ethanol for 72 hours. The resulting ethanolic extract was screened for the presence of phytochemicals. Vero cells monolayer in 96-well plate was infected with RH strain of T. gondii and treated with concentrations of the EETC, Veratrine alkaloid, and clindamycin ranging from 1.56 to 200 μg/mL. MTT assay was conducted after 24 hours to evaluate the cytotoxicity and antiparasitic activities of the EETC. Four and 24 hours treatment models were adapted to assess the infection index and intracellular proliferation of T. Results The study revealed that the EETC had no cytotoxic effects on Vero cells with IC50 = 179 μg/mL, as compared to clindamycin (IC50 = 116.5 μg/mL) and Veratrine alkaloid (IC50 = 60.4 μg/mL). The EETC had good anti-toxoplasma activities with IC50 = 6.31 μg/mL in comparison with clindamycin (IC50 = 8.33 μg/mL) and Veratrine alkaloid (IC50 = 14.25 μg/mL). The EETC caused more than 70% and 80% reduction in infection index and intracellular proliferation in both treatment models, respectively. Conclusion This in vitro study showed that the EETC contains promising phytochemicals effective against T. gondii and safe to the host cells.

[1]  Ryo Takano,et al.  Screening of a library of traditional Chinese medicines to identify anti-malarial compounds and extracts , 2018, Malaria Journal.

[2]  W. Pavanelli,et al.  Pravastatin and simvastatin inhibit the adhesion, replication and proliferation of Toxoplasma gondii (RH strain) in HeLa cells. , 2017, Acta tropica.

[3]  W. Sullivan,et al.  TgPRELID, a Mitochondrial Protein Linked to Multidrug Resistance in the Parasite Toxoplasma gondii , 2017, mSphere.

[4]  J. S. Doggett,et al.  Drugs in development for toxoplasmosis: advances, challenges, and current status , 2017, Drug design, development and therapy.

[5]  A. Daryani,et al.  A Systematic Review of In vitro and In vivo Activities of Anti-Toxoplasma Drugs and Compounds (2006–2016) , 2017, Front. Microbiol..

[6]  K. Dzitko,et al.  Human toxoplasmosis-Searching for novel chemotherapeutics. , 2016, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[7]  Chunmei Jin,et al.  Antiparasitic effects of oxymatrine and matrine against Toxoplasma gondii in vitro and in vivo. , 2016, Experimental parasitology.

[8]  W. Ahmad,et al.  Tinospora crispa (L.) Hook. f. & Thomson: A Review of Its Ethnobotanical, Phytochemical, and Pharmacological Aspects , 2016, Front. Pharmacol..

[9]  Xing-Quan Zhu,et al.  Impact of environmental factors on the emergence, transmission and distribution of Toxoplasma gondii , 2016, Parasites & Vectors.

[10]  Z. Hassan,et al.  Apoptotic activity and anti-Toxoplasma effects of artemether on the tachyzoites and experimental infected Vero and J774 cell lines by Toxoplasma gondii , 2016, Indian journal of pharmacology.

[11]  R. Anandalakshmi,et al.  Qualitative and quantitative analysis of phytochemicals in crude extract of big – Leaf mahogany (Swietenia macrophylla King.) , 2016 .

[12]  V. Titanji,et al.  Evaluation of the cytotoxic activity of extracts from medicinal plants used for the treatment of malaria in Kagera and Lindi regions, Tanzania , 2015 .

[13]  Tim Cushnie,et al.  Alkaloids: an overview of their antibacterial, antibiotic-enhancing and antivirulence activities. , 2014, International journal of antimicrobial agents.

[14]  Walaa Najm Abood,et al.  Immunomodulatory effect of an isolated fraction from Tinospora crispa on intracellular expression of INF-γ, IL-6 and IL-8 , 2014, BMC Complementary and Alternative Medicine.

[15]  P. Houghton,et al.  Anticholinesterase Inhibitory Activity of Quaternary Alkaloids from Tinospora crispa , 2014, Molecules.

[16]  S. Srichairatanakool,et al.  Antimalarial Activity of Stem Extract of Tinospora Crispa against Plasmodium Berghei Infection in Mice , 2014 .

[17]  Y. Koay,et al.  A Review of the Secondary Metabolites and Biological Activities of Tinospora crispa (Menispermaceae) , 2013 .

[18]  David E. Williams,et al.  Crude extract and purified components isolated from the stems of Tinospora crispa exhibit positive inotropic effects on the isolated left atrium of rats. , 2013, Journal of ethnopharmacology.

[19]  S. A. Fadeyi,et al.  In vitro anticancer screening of 24 locally used Nigerian medicinal plants , 2013, BMC Complementary and Alternative Medicine.

[20]  N. Ilyas,et al.  Evaluation of Phytochemical Screening and Analgesic Activity of Aqueous Extract of The Leaves of Microtrichia perotitii Dc (Asteraceae) in Mice using Hotplate Method , 2013 .

[21]  J. Chu,et al.  Antiparasitic effects of Zingiber officinale (Ginger) extract against Toxoplasma gondii , 2013 .

[22]  F. Cabrerizo,et al.  In vitro evaluation of β-carboline alkaloids as potential anti-Toxoplasma agents , 2013, BMC Research Notes.

[23]  S. Kortagere Screening for small molecule inhibitors of Toxoplasma gondii , 2012, Expert opinion on drug discovery.

[24]  R. Noordin,et al.  Alkaloids content, cytotoxicity and anti-Toxoplasma gondii activity of Psidium guajava L. and Tinospora crispa , 2012 .

[25]  N. Kavitha,et al.  In vitro Anti-Toxoplasma gondii Activity of Root Extract/Fractions of Eurycoma longifolia Jack , 2012, BMC Complementary and Alternative Medicine.

[26]  N. M. Silva,et al.  Enrofloxacin is able to control Toxoplasma gondii infection in both in vitro and in vivo experimental models. , 2012, Veterinary parasitology.

[27]  F. Robert-Gangneux,et al.  Epidemiology of and Diagnostic Strategies for Toxoplasmosis , 2012, Clinical Microbiology Reviews.

[28]  Ming-Jai Su,et al.  Hypoglycemic diterpenoids from Tinospora crispa. , 2012, Journal of natural products.

[29]  H. Ginsburg,et al.  A call for using natural compounds in the development of new antimalarial treatments – an introduction , 2011, Malaria Journal.

[30]  S. Ananvoranich,et al.  Toxoplasma gondii: inhibitory activity and encystation effect of securinine and pyrrolidine derivatives on Toxoplasma growth. , 2011, Experimental parasitology.

[31]  B. Khoo,et al.  Optimization of Toxoplasma gondii cultivation in VERO cell line. , 2010, Tropical biomedicine.

[32]  T. Boonmars,et al.  In vivo antiparasitic activity of the Thai traditional medicine plant--Tinospora crispa--against Plasmodium yoelii. , 2009, The Southeast Asian journal of tropical medicine and public health.

[33]  Jian-pin Cao,et al.  Anti-Toxoplasma gondii activity of GAS in vitro. , 2008, Journal of ethnopharmacology.

[34]  K. Saliba,et al.  Common dietary flavonoids inhibit the growth of the intraerythrocytic malaria parasite , 2008, BMC Research Notes.

[35]  M. Riou,et al.  Flavonoids and related compounds in parasitic disease control. , 2008, Mini reviews in medicinal chemistry.

[36]  J. Mead,et al.  Antiparasitic activity of flavonoids and isoflavones against Cryptosporidium parvum and Encephalitozoon intestinalis. , 2006, FEMS microbiology letters.

[37]  V. Yardley,et al.  Antitrypanosomal and Antileishmanial Activities of Flavonoids and Their Analogues: In Vitro, In Vivo, Structure-Activity Relationship, and Quantitative Structure-Activity Relationship Studies , 2006, Antimicrobial Agents and Chemotherapy.

[38]  J. Robinson,et al.  Evaluation of French Guiana traditional antimalarial remedies. , 2005, Journal of ethnopharmacology.

[39]  J. Montoya,et al.  Toxoplasmosis , 2004, The Lancet.

[40]  A. Omar,et al.  In vitro antifilarial effects of three plant species against adult worms of subperiodic Brugia malayi. , 2001, Journal of Ethnopharmacology.

[41]  S. Kojima,et al.  Antimalarial activity of extracts of Malaysian medicinal plants. , 1999, Journal of ethnopharmacology.