ANTISICKLING ACTIVITY EVALUATION OF FRACTIONS OBTAINED FROM WHOLE EXTRACTS
OF NEWBOULDIA LAEVIS P. BEAUV (BIGNONIACEAE)
Original Article
AFFO DERMANE
a,b*
, KAFUI KPEGBA
b
, KOSSI METOWOGO
c
, M. KOSSI JOPPA
c
, A. KODJO AKLIKOKOU
c
a
Department of Pharmacy, Universite de Lome, Togo,
b
Laboratory of Organic Chemistry and Natural Substances, Universite de Lome,
Togo,
c
Received: 03 Oct 2018 Revised and Accepted: 20 Dec 2018
Laboratory of Physiology-Pharmacology, Universite de Lomé, Togo
Email: dermaneaffo@yahoo.fr
ABSTRACT
Objective: This study aims to evaluate the fractions from whole extracts of roots and stem barks of Newbouldia laevis and their effect on sickling.
Methods: Hydroethanolic extracts of stem barks and roots of Newbouldia laevis were fractionated by the technics of cold precipitation in ethanol.
The fractions obtained after phytochemical screening were subjected to Emmel test to evaluate their anti-sickling activity. Active fractions were
tested for DPPH and AAPH assay (AAPH induced membrane lipoperoxidation and evaluation of reduction of hemolysis).
Results: Two fractions were obtained from each whole extract: supernatant and pellet fractions. Supernatants fractions obtained from whole roots
barks extract and stem barks extract at a concentration of 30 mg/ml reduced sickling up respectively to 7% and 10% against 86% for the control.
Pellets fractions obtained from the both extracts induced coagulation of SS blood at 30 mg/ml against 86% for the control.
Conclusion: Supernatants fractions of hydroethanolic whole extract of roots and stem backs of Newbouldia laevis promise as the potential source of
active molecules against sickle cell disease.
Keywords: Sickling, Supernatant fraction, Newbouldia laevis, Sickle cell disease, SS blood
© 2019 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)
DOI: http://dx.doi.org/10.22159/ijpps.2019v11i2.30112
INTRODUCTION
Sickle cell disease is a genetic hemoglobinopathy of autosomal
recessive inheritance affecting the β chain of the protein. It is due to
an amino acid permutation which induces a change in the
confirmation of the chain (qualitative anomalies) that allows it to
polymerize when deoxygenated. According to Aygun and Odame [1],
there are Hemoglobin A (Hb A), natural, Hemoglobin S (Hb S) caused
by the β 6 (A 3) mutation where glutamic acid (Glu) is substituted by
valine (Val) and Hemoglobin C (Hb C) caused by the β6 (A 3
While healthy carriers of Hb S/HbA (Hb AS) or Hb AC have few
clinical manifestations, the impact is important for HbSS
homozygotes. The polymerization of hemoglobin will induce a
change in conformation of the red blood cell (passage from a
biconcave to a sickle shape), reducing both its life and its functional
capacity. The clinical signs that will appear initially are chronic
anemia and high susceptibility to infections and then complications
may arise due to alteration of the blood circulation inducing
cerebrovascular accidents caused by vaso-occlusive crisis, hypoxia
coupled with pulmonary infiltration resulting in acute thoracic
syndrome, aplastic anemia that can lead to death, stunting and
various complications in adulthood [2].
)
mutation where glutamic acid (Glu) is replaced by lysine (Lys).
In 1994, the World Health Organization (WHO) estimated that 120
million people (2.3% in the world) had the sickle-cell mutation. On
the other hand, in Africa, this rate can reach 40% [3].
Available treatments are limited. The marrow transplantation
remains at present the only curative treatment but it is not possible
in developing countries where the care consist to prevent
complications and relieve pains [4].
Treatment with hydroxyurea to reduce the risk of stroke, acute
thoracic syndrome and also reduce inflammatory molecules [5]
remains expensive for most patients. Blood transfusion is also
proposed with some measures that are difficult to implement in our
developing countries [6]. For these reasons, sickle cell patients in
sub-Saharan Africa resort to medicinal plants that have proved their
effectiveness. Plants have played a significant role in maintaining
human health and improving the quality of human life for thousands
of years. This resulted in an intense global search for plant extracts
and their constituents for health care [7].
In this case, our previous studies and that of Joppa et al. [8]
demonstrated an anti-sickling effect of roots and stem barks extract
of Newbouldia laevis. The objective of this study was to potentiate
the anti-sickling effect of total extracts in the fraction(s) of this
extract which led to identifying the bioactive compounds. This study
was performed to evaluate the effects of fractions from total extracts
on sickling.
MATERIALS AND METHODS
Materials
Plant material
Roots and stem barks of Newbouldia laevis were harvested at Tsévié,
an area that is situated at 35 km at the northern part of Lomé in
December 2014. The plant had been identified at the Botany
Department of the Faculty of Sciences of the University of Lomé where
it is registered in the herbarium and kept under the number 233.
Blood samples
The SS blood samples used was provided by the hematology
laboratory of the CHU Campus. The control of hemoglobin status has
been proven by the electrophoresis method of hemoglobin. Only the
hematologist after consultation determine the samples to be used.
These samples are taken from people who had come for
hematological consultation at CHU Campus and who had not
undergone blood transfusions for 120 d, in EDTA tubes and used
fresh. These people (at least 3) should not have been on anti-sickling
and antioxidant treatment for 7 d.
Animal material
Healthy Wistar rats weighing 90 g to 100 g were used to a collected
blood sample for evaluation of active fractions antioxidant effect.
International Journal of Pharmacy and Pharmaceutical Sciences
ISSN- 0975-1491 Vol 11, Issue 2, 2019