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This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License
Article
Author(s)
Shaba Peter, Sahab Dey, Bhanuprakash Veerakyathappa, Singh Raj Kumar and Chaudary Paulad
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DOI:10.17265/2328-2150/2016.10.004
Affiliation(s)
ABSTRACT
Emblica officinalis (E. officinalis) dried fruits were
evaluated for its antitrypanosomal activity and cytotoxic effects. Vero cell
line maintained in DMEM (Dubecco’s Modified Eagle Medium) and incubated with Trypanosoma
evansi for more than 12 h. MPE was added to the Vero cell culture medium at
different concentrations (250~1,000 µg/mL) with
trypanosomes concentration (1 × 106 trypanosomes/mL in each ELISA plate well) and incubated at appropriate
conditions for 72 h. In-vitro cytotoxicity of MPE
of E. officinalis was determined on
Vero cells at concentrations ((1.56~100 µg/mL). Acute toxicity and in-vivo infectivity tests
were done in mice. Obtained MPE of E.
officinalis underwent process of purification via column chromatography,
preparative chromatography and HPLC (higher performance liquid chromatography) with bioassay at
different strata on Alsever’s medium. In-vivo assay for
trypanocidal activity, MPE and PPFs (partially purified fractions) of E. officinalis with two sets of mice,
each mouse was inoculated with 1 × 104/mL of
trypanosomes and treated (48 h post inoculation) at concentrations (12.5, 25,
50, 100 and 200 mg/kg body weight) were administered at dose rate of 100 µL per
mouse via intraperitoneal route (in treating parassitemic mice) to different
groups of mice, 6 mice per concentration. HPLC of partially purified fractions
of E. officinalis was carried out with mobile phase of acetonitrile:
water (40:60) in gradient mode. In vitro, MPE induced immobilization and
killing of the parasites in concentration-time dependent manner. Significant
reduction of trypanosomes counts from concentration of 250 µg/mL and complete
killing of trypanosomes at 5th hour of observation,
which was statistically equivalent to 4th hour of Diminazine Aceturate (Berenil), standard
reference drug used. HPLC of the partially purified fractions revealed two
major prominent peaks at retention time of 1~4 min. In vivo, both MPE and
PPFs of test material did prolong lives of mice by 6~9 days but could not cure them. At concentration of 2,000 kg/kg body weight
of MPE in acute test, all mice survived. For in-vivo infectivity test, mice injected with immobilized trypanosomes developed
parasitemia and died while, the other group survived. MPE, PPFs and Diminazine
Aceturate were toxic to Vero cells at all concentrations exception of 1.56,
1.56~3.13 and 1.56~6.25 µg/mL,
respectively. From this report, PPFs of E.
officinalis dried fruits demonstrated potential pathway for a new
development of trypanocide in near future if additional investigations are put
in place.
KEYWORDS
Emblica officinalis dried fruits, in vitro and in vivo partially purified fractions trypanocidal activity, Trypanosoma evansi, in vitro cytotoxicity.
Cite this paper
Peter, S., et al. 2016. “Therapeutic Activity of Partially Purified Fractions of Emblica officinalis (Syn. Phyllanthus emblica) Dried Fruits against Trypanosoma evansi.” Journal of Pharmacy and Pharmacology 4 (10): 546-558.
References