Medicinal Plants:
Phytochemistry, Pharmacology
and Therapeutics
iii
Medicinal Plants:
Phytochemistry, Pharmacology
and Therapeutics
Volume 1
Editor-in-Chief
V.K. Gupta
Editors
G.D. Singh
Surjeet Singh
A. Kaul
2010
iv
2010 EDITORS
ISBN 81-7035-627-X
ISBN 978-81-7035-627-1
All rights reserved. Including the right to translate or to reproduce this book or parts thereof except for brief
quotations in critical reviews.
Published by
Showroom
Laser Typesetting
Printed at
PRINTED IN INDIA
Dedicated to
Prof. C. K. Atal
Former Director, Indian Institute of Integrative Medicine, Jammu
(Erstwhile, Regional Research Laboratory, Jammu)
vii
Foreword
The twenty-nine chapters of the book Medicinal Plants: Phytochemistry, Pharmacology and
Therapeutics, Volume 1, edited by Dr. V. K. Gupta, Dr. G. D. Singh, Dr. Surjeet Singh and Dr. A. Kaul
(2009) represent enormous progress, as they cover themes currently under discussion in all of these
research fields. Some of the contributions are still based on Ayurvedic herbs and medicine, but most
are of general interest, addressing herbs with antioxidant, antilipedemic, vasodilatory, opioid,
antiinflammatory, antisickling, antimicrobial, antidiabetic antiparasitic, antimalaria or antiaging
activities. This wide spectrum of themes may be of great value for research in natural products chemistry
and biology, and to medical doctors as well.
I congratulate the editors and appreciate their efforts in bringing out such an excellent book
which will give all round readers an exciting and serious reading material and also continuing the
outstanding works for which Indian Institute of Integrative Medicine (CSIR), Jammu (Formerly Regional
Research Laboratory, Jammu) has been known.
It is a pleasure for me to recommend this volume without reservation to all researches in the field
of phytomedicine. I wish the book much success and a broad distribution.
I do hope that this book will be followed by second volume in the next years.
Dr. H. Wagner
Professor Emeritus
Centre of Pharma Research, Department Pharmacy,
University of Munich, Germany
ix
Preface
Plants have been used for alleviating human suffering from the very beginning of human
civilization, and records of the use of plants are available since about 5000 years ago. The active
principles isolated, have provided leads in the development of several life saving drugs, which are in
use today. Different civilizations developed their own indigenous system of medicines. Historically,
about two centuries ago, our medicinal practices were largely dominated by plant-based medicines.
However, the medicinal use of herbs went into decline in the West when more predictable synthetic
drugs were made commonly available. In contrast, many developing nations continued to benefit
from the rich knowledge of medical herbalism. For example Ayurvedic medicine in India, Kampo
medicine in Japan, Traditional Chinese Medicine and Unani Medicine in the Middle East and South
Asia are still used by a large majority of people.
All around the world there is talk about health for all but it has been realized that modern
pharmaceuticals are and will remain out of reach of a large proportion of the human population for
the foreseeable future. This necessitates the use of other sources of human knowledge to provide
common health benefits. Thus, herbal medicine is now regarded as important but underutilized tool
against disease. The World Health Organization (WHO) recognized this fact in the early 1970s and
encouraged governments to effectively utilize local knowledge of herbal medicines for disease prevention
and health promotion.
There is now a popular belief that allopathic drugs have serious side effects on human body. As
against the same, herbal medicines work better and provide long lasting healing effect and are without
any side effects. As such there is now a growing demand of herbal medicines and herbal therapeutic
applications. The primary health care of 70-80 per cent of the worlds population is based on the use
of medicinal plants derived from traditional systems of medicine and local health practices. During
the past few decades public interest in traditional, complementary and alternative medicine (TCAM)
and use of herbal medicines has increased dramatically in industrialized countries. Traditional
medicine has a bright future and an immense potential to extend medical relief to millions, who for
lack of resources remain deprived of it. When undesirable side effects of certain drugs have unnerved
the patients, herbal medicine is the only hope in India where 60 per cent of the population lives below
the poverty line.
This has increased the international trade in herbal medicine enormously. WHO said in 2003
that the global market for herbal medicines stood at US $ 60 billion and was growing steadily. Global
sales of herbal products including herbal medicine has already crossed 100 billion in the last five
years and is expected to exceed one trillion in the next 20 years at the present growth rate. In India, the
herbal drug market is about $ one billion and the export of plant based crude drugs is around $ 80
million.
Many pharmaceutical companies are showing interest in the production and marketing of herbal
medicines. The sales for herbal medicine products have plateaued to such an extent that these products
have become available to consumers as positive healthcare just like vitamins. Herbal medicines are in
great demand in the developed as well as developing countries for primary healthcare because of their
wide biological activities, higher safety margins and lesser costs.
Out of 20,000 plants recognized of medicinal value, only a very few are in use. Their use is not
scientifically validated much with the scientific data. Plant extracts of therapeutic relevance are of
paramount importance as reservoirs of structural and chemical diversity. A recent report reveals that
at least 120 distinct chemical substances from different plants have utility as lifesaving drugs. This
has been achieved through chemical and pharmacological screening of only 6 per cent of the total
plant species.
It is for their world wide and a sustained effort of scientists that an enormous information is
being generated and there has been a series of publications on medicinal plant researches. Based on
this rational, the present book Medicinal Plants: Phytochemistry, Pharmacology and Therapeutics Vol. 1 presents information on review/research communications received from eminent scientists
from India and abroad, providing recent and present state of the art data on therapeutic properties,
action and uses of medicinal plants in combating a number of diseases and condition for which there
is lesser satisfactory treatment in modern medicine.
It is hoped that the present volume will attract wide acceptance of phytochemists, pharmacologists,
medical personals in particular and a host of other scientists and biologists to facilitate further research
on medicinal plants.
V.K. Gupta
G.D. Singh
Surjeet Singh
A. Kaul
xi
Contents
Foreword
Preface
1.
vii
ix
1
32
51
75
S. Singh, R. Sharma, G.D. Singh, A. Kaul, A. Khajuria, P. Koul and V.K. Gupta
5.
98
131
xii
7.
155
178
Cludia Ferreira Santos, Antonia Torres Dvila Pimenta, Adriana Sousa Barros,
Victor Martins Gomes, Natlia Rocha Celednio, Marta Regina Kerntopf,
Silvnia Maria Mendes Vasconcelos, Ana Maria Sampaio Assreuy,
Elnatan B. de Souza, Mary Anne Sousa Lima, Edilberto Rocha Silveira,
Francisco Arnaldo Viana and Nilberto Robson Falco do Nascimento
9.
Comparative Effects of Soybean, Sunflower, Olive and Sugar Cane Wax Oils
and their Respective Fatty Acids in Cutaneous Inflammation
192
202
J. Gao, Y.H. Tang, L.Z. Xu, X.H. Tang and X.N. Zhao
11. Multi-Targeted Approaches for Polygenic Disorders Using Medicinal Plants:
A New Battle Against Old Adversaries
V.S. Muthusamy and B.S. Lakshmi
12. Safety Assessment of Orthosiphon stamineus Benth Methanol Leaf Extract:
Drug Interaction and Oral Toxicity Study in Rats
215
228
238
258
297
315
330
xiii
338
354
363
378
384
390
397
406
413
421
430
441
449
475
476
Pages 441447
Chapter 29
ABSTRACT
The antimalarial bioactivity of aqueous extract of stem back of Enantia chlorantha was
investigated in Plasmodium berghei infected mice. Stem bark of Enantia chlorantha was analysed
for its phytochemicals. Twenty five (25) albino mice were infected by intraperitoneal injection of
standard inoculum of chloroquine sensitive Plasmodium berghei (NK 65 strain). The animals were
randomly divided into 5 groups of 5 mice each. Group A served as the control while groups B
and C were administered 1.75 and 5 mg/kg body weight of artesunate and chloroquine
respectively. Groups D and E received 100 and 400 mg/kg of extract of E. chlorantha orally. The
results showed the presence of alkaloids saponins, phenolics, flavonoids and glycosides. There
was 100 per cent parasite clearance in the 400mg/kg extract and chloroquine groups, and 98.6
per cent clearance in the group that received 100mg/kg body weight of extract. There was no
parasite clearance in the artesunate group. There was 100 per cent mortality in the negative
control group; 40 per cent mortality in the artesunate, chloroquine and 100mg/kg body weight
extract group and 60 per cent mortality in the 400mg/kg extract group. The Mean Survival Time
for the control group was 9.0 days; artesunate 22 and chloroquine 19.8 days, while the
groups that received 100 and 400 mg/kg body weight of extract recorded 19.6 and 17.0 days
respectively.
442
Keywords: Antimalarial bioactivity, Enantia chlorantha, Stem bark, Plasmodium berghei, Mice.
Introduction
Malaria remains the major cause of morbidity and mortality in the tropical regions of the world
with over 300 million new cases reported annually (WHO, 2005). Almost 90 per cent of the deaths
from malaria occur in sub-Saharan Africa, where the vulnerable groups are children under 5 years
and the pregnant women (WHO, 1999).
WHO experts say that the number of people infected with malaria is still increasing at the rate of
about 5 per cent annually, and this has been attributed largely to increasing incidence of resistance to
antimalarial drugs formerly effective against the pathogen. Antimalarial drug resistance has become
one of the greatest challenges against malaria control. There is widespread multi-drug resistance to
common antimalarial drugs (Muregi et al., 2003; WHO, 2005).
In Africa, more than 80 per cent of the people use traditional herbal remedies for the treatment of
many ailments including malaria (Akerele, 1984; Wright and Phillipson, 1990).
Rodent plasmodia such as Plasmodium yoeli and Plasmodium berghei are commonly used as malaria
models in mice and have tremendous impact on the investigation of antimalarial activity of plant
extracts.
The need to search and develop more effective antimalarial drugs that are inexpensive and readily
available to people in the developing countries like Nigeria has necessitated this study.
Aqueous Extraction
Stem bark of the plant was air-dried to constant weight and ground into powdered form with an
electric blender (Blender/Miller III, model MS 223) Taiwan, China. Aqueous extract was prepared as
described previously (Akanji and Adesokan, 2005).
Phytochemical Analysis
A portion of the stem powder was subjected to phytochemical analysis, using standard chemical
tests as described earlier (Odebiyi and Sofowora, 1978; Trease and Evans, 1989).
Experimental Animals
Albino mice, weighing 20-25g, were obtained from the small Animal Holding Unit of the
Department of Pharmacology, College of Health Sciences, University of Ilorin, Ilorin, Nigeria. The
443
animals were housed in wire mesh cages under standard conditions, and the study conducted in
accordance with the recommendations from the declaration of Helsinki on guiding principles in the
care and use of animals.
Malaria Parasite
Plasmodium berghei (chloroquine sensitive NK 65 strain) was obtained from the Institute for
Advanced Medical Research and Training (IMRAT), College of Medicine, University of Ibadan, Ibadan,
Nigeria.
Animal Groupings
The animals were randomly divided into 5 groups of 5 mice each, after confirmation of
parasitaemia 72 h post-inoculation. Group A, (control) was left untreated but administered appropriate
volume of distilled water. Group B received artesunate orally at a dose of 1.75mg/kg body weight
daily for 4 days and group C 5mg/kg body weight of chloroquine base for the same period. Groups D
and E were administered aqueous extract of Enantia chlorantha through oropharyngeal canula at the
doses of 100 and 400mg/kg body weight respectively.
x 100
x 100
444
Results
Phytochemical analysis
Qualitative and quantitative screening of the components of the plant stem bark yielded the
phytochemicals shown in Table 29.1. The phytochemicals that were present included Alkaloids (46.26
per cent), Saponins (26.82 per cent), Phenolics (18.77), Flavonoids (6.71 per cent) and Glycosides (1.44
per cent).
Table 29.1: Qualitative and Quantitative Phytochemical Analysis of Enantia chlorantha
Phytochemical
Qualitative
Quantitative
Mg
Percentage
Phenolics
++
1.120.00
18.77
Flavonoids
0.400.02
6.71
Alkaloids
++
2.760.02
46.26
Glycosides
0.0860.01
1.44
Saponins
++
1.600.02
26.82
Tannins
Nd
0.00
0.00
Phlebotanins
Nd
0.00
0.00
Steroids
Nd
0.00
0.00
Percentage Parasitaemia
Estimation of percentage parasitaemia at the end of 28 days showed the results in Table 29.2.
There was no parasitaemia in the blood of mice in the chloroquine group and the group that received
400mg/kg body weight of extract of Enantia chlorantha. Less than 2 per cent parasitaemia was found in
the blood of mice that received 100mg/kg body weight of the extract. There was high parasitaemia of
55 per cent in the artesunate group.
Table 29.2: Percentage Parasitaemia (Day 28) in Experimental Groups Following Administration of
Standard Antimalarial Drugs and Extracts of Enantia chlorantha
Treatment Groups
Percentage Parasitaemia
Artesunate (1.75mg/kg)
55
Chloroquine (5mg/kg)
E. chlorantha (100mg/kg)
<2
E. chlorantha (400mg/kg)
Percentage Mortality
At the end of the observational period, 100 per cent mortality was recorded for the untreated
control group (Table 29.3). Forty percent (40 per cent) mortality was recorded for the artesunate and
445
chloroquine groups and the extract group that received 100mg/kg body weight dose, but 60 per cent
in the group that received 400mg/kg body weight of the extract.
Table 29.3: Percentage Mortality (Day 28) in Experimental Groups Following Administration of
Standard Antimalarial Drugs and Extracts of Enantia chlorantha
Treatment Groups
Percentage Mortality
Control
100
Artesunate (1.75mg/kg)
40
Chloroquine (5mg/kg)
40
E. chlorantha (100mg/kg)
40
E. chlorantha (400mg/kg)
60
MST (Days)
Control
9.00.8
Artesunate (1.75mg/kg)
22.02.1
Chloroquine (5mg/kg)
19.81.8
E. chlorantha (100mg/kg)
19.61.6
E. chlorantha (400mg/kg)
17.01.3
Discussion
Results from this study showed that aqueous extract of Enantia chlorantha possess potent
antimalarial activities that were comparable to that of chloroquine, while the observed antimalarial
activities were not dose dependent.
The stem bark of E. chlorantha consisted of preponderant alkaloids and phenolics, both of which
may be responsible for the pharmacologic activity of the extract. Earlier workers have shown that
isolated alkaloid, 9-methoxycanthin-6-one displayed higher antimalarial activity against Plasmodium
falciparum Gombak A isolate, when compared with chloroquine (Chan et al., 2004). In addition, potent
antimalarial agents, raphidecurperoxin and polysyphorin, were isolated from the Vietnamese medicinal
plants, Rhaphidophora decursiva (Zhang et al., 2001). The extracts of Nigella sativa (Black seed), contained
different classes of alkaloids that were believed to block protein synthesis in Plasmodium falciparum
(Abdulelah and Zainal-Abidin, 2007).
In addition, phenolics, which are known to possess antiparasitic, anticarcinogenic,
antiinflammatory and immunomodulatory effects, may also play a significant role in the antimalarial
activity of the extract (Abdulelah and Zainal-Abidin, 2007).
446
Consistent with this concept, different extracts of Enantia chlorantha have been reported to exert
antimicrobial activities, including antibacterial (Agbaje and Onabanjo, 1991; Adesokan et al., 2007).
The antioxidant effect of plant alkaloids may represent another mechanism that contributed to its
antimalarial activity. Antioxidant components might inhibit nitric oxide (NO) production in
macrophages which will lead to increased degradation of tryptophan and thereby starve the parasite
of an essential amino acid leading to its death (Daubener, 1999; Mahmoud et al., 2003).
The antimalarial activity as demonstrated by the percentage parasitaemia in the groups that
received the extracts compared favourably with that of chloroquine. The percentage mortality of the
animals in the group that received 100mg/kg body weight of the extract was similar to those of the
artesunate and chloroquine groups and better than the negative control group.
Further more, the mean survival time of 19.6 days in the group that received 100mg/kg body
weight was similar to 19.8 days for the chloroquine group and compared well with 22 days of the
group that received artesunate. Even MST of 17 days in the group that was administered 400mg/kg
body weight of the extract has proven that the extract possess potent antimalarial activity. Survival of
experimental animals beyond 12 days is regarded as significant activity (Peters, 1980; Obih and
Makinde, 1985; Abosi and Raseroka, 2003; Ajaiyeoba et al., 2006).
The active principles responsible for these antimalarial activities are yet to be identified but the
results from this study have largely justified its use in folklore medicine for malaria treatment in
Africa.
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