ABSTRACT The effect of the extracts of Ruellia Praetermissa Schweinf. ex Lindau. on hemoglobin (Hb), Hematocrit (Hct), mean corpuscular hemoglobin (MCH), mean corpuscular volume (MCV). mean corpuscular hemoglobin concentration (MCHC), red blood cell count (RBC), was investigated in 50 Pregnant women attending prenatal clinic in Belo maternity. The women were assigned to 5 groups of 10 women per group. The first group was the control and the other 4 were the experimental groups. The control was administered daily, 0.5 ml of saline solution while the experimental groups were administered daily oral doses of the plant extract in concentrations of 200 mg/kg, 400 mg/kg, 800 mg/kg and 1,600 mg/kg respectively for 16 days. Blood samples were collected on the 17 th day and analyzed. The extracts contain flavonoids aglycones (luteolin and apigenin) and their respective, glycosides and a high concentration of triterpenes (campesterol, stigmasterol, -sitosterol, lupeol) and iridoid glycosides. It was also found to be rich in vitamins and minerals. The extracts increased the values in a dose dependent manner Hb (P &lt; 0.05), RBC (P &lt; 0.05), hematocrit (P &lt; 0.05). It however showed no remarkable effect on the values of MCH and MCHC (P &gt; 0.05) but with a dose depending decreasing effect on MCV (P &lt; 0.05). The active principles of this plant drug stimulate erythropoiesis which leads to increase in circulating RBCs with slightly microcytic sizes (MCV), normochromic weight, (MCH) normochromic hemoglobin contents (MCHC). The result of this study thus supports the traditional use of Ruellia Praetermissa in pregnancies threatened with miscarriage and as a remedy for anemia

Njunda L Anna

Salah A Martin

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Global Journal of Research on Medicinal Plants & Indigenous Medicine 
 
 
Original Research Article 
EFFECT OF RUELLIA PRAETERMISSA EXTRACTS ON ERYTHROPOIESIS 
IN PREGNANT WOMEN 
Salah A. Martin
1*
, Njunda L. Anna
2
 
 
1
Department of Zoology and Animal Physiology, Faculty of Science, University of Buea, P.O. Box 63 Buea, 
Cameroon.  
2
Department of Medical Laboratory Sciences, Faculty of Health Science, University of Buea, Cameroon. 
 
*
Corresponding Author: E-mail: salahmartin@yahoo.fr 
 
Received: 14/06/2012; Revised: 22/07/2012; Accepted: 24/07/2012 
  
ABSTRACT 
The effect of the extracts of Ruellia Praetermissa Schweinf. ex Lindau. on hemoglobin (Hb), 
Hematocrit (Hct), mean corpuscular hemoglobin (MCH), mean corpuscular volume (MCV). mean 
corpuscular hemoglobin concentration (MCHC), red blood cell count (RBC), was investigated in 50 
Pregnant women attending prenatal clinic in Belo maternity.  The women were assigned to 5 groups 
of 10 women per group.  The first group was the control and the other 4 were the experimental 
groups.  The control was administered daily, 0.5 ml of saline solution while the experimental groups 
were administered daily oral doses of the plant extract in concentrations of 200 mg/kg, 400 mg/kg, 
800 mg/kg and 1,600 mg/kg respectively for 16 days.  Blood samples were collected on the 17
th
 day 
and analyzed. The extracts contain flavonoids aglycones (luteolin and apigenin) and their respective, 
glycosides and a high concentration of triterpenes (campesterol, stigmasterol, -sitosterol, lupeol) and 
iridoid glycosides. It was also found to be rich in vitamins and minerals. The extracts increased the 
values in a dose dependent manner Hb (P < 0.05), RBC (P < 0.05), hematocrit (P < 0.05). It however 
showed no remarkable effect on the values of MCH and MCHC (P > 0.05) but with a dose 
depending decreasing effect on MCV (P < 0.05). The active principles of this plant drug stimulate 
erythropoiesis which leads to increase in circulating RBCs with slightly microcytic sizes (MCV), 
normochromic weight, (MCH) normochromic hemoglobin contents (MCHC). The result of this 
study thus supports the traditional use of Ruellia Praetermissa in pregnancies threatened with 
miscarriage and as a remedy for anemia.  
KEYWORDS: Ruellia Praetermissa, Pregnant Women, Blood 
 
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INTRODUCTION 
Gestational problems are a potential source 
of mortality for both gestation mothers and 
their developing foetuses in the underdeveloped 
world in general and Africa in particular. There 
is great need for effective safe prenatal drugs 
from natural sources which are available and 
affordable with respect to modern medicines.  
Plant drugs have effects haematological 
parametrs (Nnamdi et. al., 2012) which 
influence reproductive health. 
Ruellia praetermissa Schweinf. ex Lindau., 
is a wild herb and indigenous to central and 
south eastern Asia and also widespread in 
tropical and subtropical Africa. In the North 
West Region of Cameroon it enjoys a folk 
reputation as blood and pregnancy medicine. In 
other south Eastern African countries; it is 
widely applied to relieve pain (Gelfand et al., 
1985).  The Plant drug exerts estrogenic and 
cholinergic effects (Salah et al., 2002). The 
extracts regularize pregnancies threatened with 
miscarriages in early stages. This is due to its 
ability to mimic 17β-estradiol. It stimulates the 
growth of the uterine endometrium.  This is by 
the proliferation and the development of the 
cells of the uterine endometrium as it up-
regulates estrogen, luteinizing hormone LH and 
progesterone receptors on the uterine muscles 
at the beginning of gestation and to excite the 
uterine myometrium at term. (Salah et al., 
2002). In addition, the extracts of this plant 
drug has a stimulatory effect on the motility of 
the gastrointestinal tract (Salah et al, 2000) and 
antihypertensive effect by the inhibition of 
Angiotensin-converting enzyme (ACE) activity 
(Salah et al, 2001). These biological effects are 
of particular interest since high blood pressure 
and indigestion frequently characterize 
gestation period. This herbal drug is rich in 
flavonoids such as luteolin, quercetin and 
apigenin. (Wagner and Bladt, 1998). The 
favonoids also have an antispasmodic effect on 
uterine smooth muscle. (Salah 2001). The plant 
extract has 5-Lipoxygenase inhibition and 
antisplasmodic effects (Salah, 1999).  The plant 
drug promotes implantation, and stabilizes the 
uterine endometrium in female rats (Salah and 
Wagner, 2009). The present study is aimed at 
finding out the scientific basis of the use of 
Ruellia praetermissa as a prenatal herbal drug 
in the North-Western region of Cameroon. 
MATERIALS AND METHODS 
Plant Material 
The plant material was collected in Belo, 
North West Province of Cameroon in 
September 2011. The specimen was verified 
and authenticated as the one earlier identified 
by Kofany of the Cameroon National 
Herbarium Obili Yaounde under the voucher 
specimen number 43596 deposited in 1996. 
Extraction and sample preparation 
Sun dried leaves of the plant were 
pulverized and 250 g was extracted using the 
sohxlet for 12 h in each case progressively with 
2 l of n-hexane, chloroform, ethyl acetate, and 
methanol. The extracts (160, 140, 250 and 
300 mg, respectively) were recovered by 
rotavaporization. The chaffs were boiled in 
water at 85
o
C for 6 h and 225 mg of extract was 
recovered by lyophilization. The procedure was 
repeated 20 times to obtain the plant drug 
necessary for the whole investigation. 
Plant Drug Analysis 
a) TLC 
The ethyl acetate extract (5 mg) was co-
chromatographed with flavonoid test samples 
(rutin, chlorogenic acid, hyperoside and 
isochlorogenic acid), luteolin and luteolin-7-
glucoside, quercetin, isoquercetin, delphinidin 
and caffeic acid using ethyl acetate-formic acid 
–glacial acetic acid-water (100:11:11:26) as the 
mobile phase and precoated silica gel 60 F254 
(20 x 20 thickness 0.25 mm Merck, Darmstadt, 
Germany) as the stationary phase. The plate 
was first observed at UV254 nm then with 
natural products-polyethylene glycol reagent 
(NP/PEG) and evaluated at UV366 nm. 
b) HPLC 
This was carried out with an HP 1090. A 
liquid chromatography and an HP 1040 
photodiode array with a Hewlett Packard 
detector as a LiChrospher 100 RP 18 (5 µm) 
column 125 x 4 mm (Merck, Darmstadt, 
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Germany), and a precolumn LiChrospher RP-
18 4-4 mm (5 µm) (Merck, Darmstadt, 
Germany) with detection at wavelength 
210 nm, 254 nm, 280 nm and 366 nm. The 
mobile phase used for the separation was 
HPLC graded water (Solvent A) and 
acetonitrile (Solvent B), all acidified with 
33 µL of phosphoric acid (85%). It was started 
at 10% of solvent B and a linear gradient of 
10–30% MeCN during 20 min for a total run of 
30 min, at a flow of 1 mL/min, and a volume of 
12.5 µL of 1 mg/mL of ethyl acetate extract of 
R. praetermissa was injected. 
Experimental Subjects 
The study was conducted in Boyo, and 
Bamenda in the North West Region of 
Cameroon from December 2011 to April 
2012.  A total of 50 pregnant women between 
20 and 40 years of age attending routine 
prenatal clinics in Belo Maternity Center were 
recruited for the study. They gave their 
informed consent to participate in the study 
which was approved by the ethics commission 
of our institution. All participants were 
interviewed to assess their physical wellness, 
nutritional factors, gestational history and 
hematological records. The women were 
assigned to 5 groups of 10 each.  The first 
group was the control which received daily 
doses of 0.5 ml of saline solution.  The second, 
third, fourth and fifth groups received 
200 mg/kg, 400 mg/kg, 800 mg/kg and 
1,600 mg/kg respectively in daily oral doses for 
sixteen days.  Blood samples (0.5 ml) were 
obtained on the 17
th
 day from each of the 
women. This was done with needles and 
syringes in 10 ml tubes using venous 
puncture by drawing specimens from a 
superficial vein in the antecubital fossa of 
the arm. Ethylenediamine tetraacetic acid 
(EDTA) was used to prevent blood from 
clotting. These samples were analyzed in the 
Diagnostic Laboratory in in Bamenda for the 
determination of blood parameters under 
investigation: Hematocrit (Hct), Hemoglobin 
Concentration (Hb), and Red blood Cell Counts 
(RBC) 
 
Blood Analysis 
The Hct was measured by the percentage of 
the total blood volume that is made up of 
RBCs. The height of the RBCs column was 
measured after centrifugation and compared to 
the column of the total whole blood. The ratio 
of the height of the RBC column compared 
with the original total blood column was 
multiplied by 100% and that was recorded as 
the Hct value. The haemoglobin value and the 
red blood cell count were done by an 
automated cell counter. The values of RBC, 
hematocrit, and haemoglobin test were used to 
calculate the RBC indices (MCV, MCH and 
MCHC). 
Statistical Analysis 
Data was analysed using Sigma Plot 11 for 
Windows and the significance was set at the 
5% level. The results were expressed as 
mean ± SEM, differences between means 
analysed using Student’s‘t’ test.  P values of 
0.05 or less and 0.001 were taken as being 
statistically significant. 
RESULTS AND DISCUSSION 
The results of the chemical analysis of the 
four extracts revealed that the plant drug 
contains flavonoids aglycones (luteolin and 
apigenin) and their respective, glycosides. The 
extract also contains a high concentration of 
triterpenes (campesterol, stigmasterol, β-
sitosterol, lupeol) and iridoid glycosides. 
The effect of various doses of the extract of 
R. Praetermissa on the mean hematological 
parameters are shown in Tables 1 & 2 below. 
Hematocrit (Hct):    Increasing the dosage 
concentration of the plant drug had an effect of 
increasing the Hematocrit (Hct) in all the 
treatment groups. Hematocrit (Hct) value at 
200 mg/kg 43.90 ± 2.06 % was however not 
significantly different from that of the control 
group 43.35 ± 1.82 %.  At high doses of the 
extract, 800 mg/kg and 1,600 mg/kg, there was 
a significant increase from 47.75 ± 1.5% to 
48.25 ± 1.49 % respectively (P < 0.05). 
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Hemoglobin Concentration (Hb): Increase in 
hemoglobin concentration occurred with 
increased concentration of applied extract.  The 
hemoglobin value of 13.26 ± 1.5 g/dL of the 
control group increased to 15.22 ± 0.19 g/dL at 
800 mg/kg and 16.10 ± 0.07 g/dL at 
1,600 mg/kg respectively.  All these values are 
significantly different from the control group 
(P < 0.05).  Significant change in hemoglobin 
concentration also occurred between the two 
high doses i.e. 800 and 1,600 mg/kg.  
Red Blood Cell Count: Women that received 
200 and 400 mg/kg body weight of the extract 
had significantly lower red cell value P < 0.05 
than those who received 800 and 1,600 mg/kg 
body weight of the extract.  There was no 
significant difference between the red cell 
count of those that received 200 mg/kg dose of 
extract and the normal saline in the control 
group. 
 
Table 1: Effect of the Extract of Ruellia Praetermissa on the Hemtaocrit (Hct) haemoglobin 
concentration and red blood cell count in the pregnant women blood 
Treatment with 
Ruellia Extract 
(Hct) (%) ± 
S.E. 
Hb(g/dL) ± S.E. RBC(million/mm
3
) 
± S.E. 
0.5 ml Saline  43. 35 ± 1.82 13.26 ± 1.5 4.10 ± 0.23 
200 mg/kg 43.90 ± 2.06 13.34 ± 1.7 4.34 ± 0.18 
400 mg/kg 46.62 ± 1.60   14.75 ± 0.14 4.80 ± 1.03 
800 mg/kg 47.75 ± 1.50   15.22 ± 0.19 5.33 ± 0.03 
1600 mg/kg 48.25 ± 1.49   16.10 ± 0.07 5.53 ± 1.1 
Table 2: Effect of the Extract of Ruellia Praetermissa on the Blood indices (mean corpuscular 
volume [MCV], Mean corpuscular haemoglobin [MCV], and Mean corpuscular haemoglobin 
concentration [MCHC]). 
Treatment with Ruellia 
Extract 
MCV ± S.E. 
(mm
3
) 
MCH ± S.E. 
(Pg) 
MCHC ± S.E. (g/dL) 
0.5 ml Saline 105.68 ± 2.92 32.34 ± 1.4 30.58 ± 1.1 
200 mg/kg 101.15 ± 0.50 30.73 ± 1.73 30.38 ± 1.7 
400 mg/kg 96.26 ± 1.06 36.56 ± 0.25 31.63 ± 1.5 
800 mg/kg 89.58 ± 1.72 28.55 ± 0.53 32.87 ± 2.43 
1600 mg/kg 87.25 ± 1.35 29.11 ± 1.39 33.36 ± 1.14 
 
MCV, MCH AND MCHC 
Women in the control group who received 
only normal saline had significantly higher 
MCV values (P < 0.05) than those recorded for 
all the groups of women who received the 
extract in various doses.  The MCV values 
decreased in the dose dependent manner. The 
MCH and MCHC showed no significant effect 
(P > 0.05) as the plant drugs were administered 
in different concentrations.  
 
 
The result of this study showed that the 
aqueous extract of the leaf of Ruellia 
Praetermissa increased the overall hemoglobin 
concentration, red cell count and packed cell 
volume in Pregnant women in a dose dependent 
manner. Extract of sorghum has been 
demonstrated to present similar effects on rats. 
(Ogwumike, 2002).  However, during 
pregnancy, there is a decrease in hematocrit, 
red blood cell count and hemoglobin values due 
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to increase ECF volume that increase plasma 
volume resulting in hemodilution 
(Sembulingam and Prema, 2010).  This 
presents a potential source of anaemia for the 
gestation mother which can result in 
miscarriage. Gestational state also causes 
physiological polycythemia due to the 
increased emotional conditions and demand for 
proton buffering. Hypoxia due to the high 
demand of respiratory gases for both the 
gestation mother and the developing fetus also 
stimulates erythropoietin secretion (Strand, 
1998). Erythropoietin or erythrocyte 
stimulating factor is the main hormone 
responsible for erythropoiesis in bone marrow, 
liver and spleen. The plant extract inhibits 
Angiotensin converting enzymes (ACE) 
activity and consequently stimulates the 
secretion of erythropoietin by peritubular 
capillaries of the nephron.(Vander et. al., 
1994). Ruellia is a vegetable rich in Vitamin 
B12 (cyanocobalamin) and minerals such as 
Iron and copper. (Duke, J., 2010). 
Cyanocobalamin is the major maturation factor 
necessary for erythropoisis, iron is necessary 
for the formation of the heme part of the 
haemoglobin and copper is important for the 
absorption of iron from the gastrointestinal 
tract. (Sembulingam and Prema, 2010). The 
iridoid glycosides isolated from this plant drug 
exert an inotropic effect of the myocardial 
muscles (Salah, 1999) and hence increase the 
cardiac output during pregnancy to satisfy the 
demands of the developing fetus (Constanzo, 
2004). Increase cardiac output increases 
erythropoietic activity resulting in more than 
usual number of erythrocytes. The active 
principles of this plant drug stimulate 
erythropoiesis which leads to increase in 
circulating RBCs with slightly microcytic sizes 
(MCV), normochromic weight, (MCH) 
normochromic hemoglobin contents (MCHC). 
The increase in numbers of erythrocytes 
containing the same amount and concentration 
of haemoglobin will fulfil the physiological 
needs of the pregnant mother with the growing 
fetus. 
CONCLUSION 
This study suggests that administration of 
Ruellia praetermissa enhances the process of 
erythropoiesis in gestation mothers and hence 
counters the anemic effects of pregnancy in a 
dose dependent manner. In combination with 
our previous studies on the effects of Ruellia on 
ovulation, growth of the uterine endometrium, 
implantation and the estrogenic effects. These 
findings provide the pharmacological basis for 
the traditional use of this plant for prenatal care 
in the North West Region of Cameroon. 
ACKNOWLEDGEMENT 
The authors would like to express their 
gratitude to the Health Board of the Belo 
Maternity, the ethical committee for the 
administrative and ethical clearances and all the 
gestation mothers who accepted to participate 
in this study. 
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Source of Support: Nil                             Conflict of Interest: None Declared 
 


EFFECT OF RUELLIA PRAETERMISSA EXTRACTS ON ERYTHROPOIESIS IN PREGNANT WOMEN

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EFFECT OF RUELLIA PRAETERMISSA EXTRACTS ON ERYTHROPOIESIS IN PREGNANT WOMEN

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