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Journal of Ethnopharmacology 116 (2008) 2126

Acute and subchronic oral toxicity of Galega ofcinalis in rats


Hamid R. Rasekh , Pardis Nazari, Mohammad Kamli-Nejad, Leila Hosseinzadeh
School of Pharmacy, Shaheed Beheshti University of Medical Sciences, P.O. Box 14155-6153, Tehran, Iran
Received 6 August 2006; received in revised form 7 October 2007; accepted 20 October 2007
Available online 30 October 2007

Abstract
Galega ofcinalis L. (Papilionaceae) is widely used in folk medicine as antidiabetic or for increasing lactation. There is a little information
about its possible toxicity. In this study, acute and subchronic toxicity of aerial parts of Galega ofcinalis in Wistar rats have been evaluated. For
the acute toxicity study, the animals received orally four different single dose of plant suspension and were kept under observation for 14 days.
The results indicated that LD50 of Galega ofcinalis is higher than 5 g/kg. In the subchronic study, 48 rats were divided into four groups and were
fed a diet containing 0%, 0.15%, 1.5% and 3% (w/w) of Galega ofcinalis. After 90 days blood and tissue samples were taken for hematological,
biochemical and histopathological determinations. An increase in serum levels of cholesterol, creatine phosphokinase, lactate dehydrogenase and
total and conjugated bilirubin was observed. Some parameters such as calcium, albumin, albumin/globulin ratio, hematocrit, WBC and platelet
counts were decreased. In microscopic examination, sinusoidal congestion in liver and alveolar hemorrhage was observed. Other parameters
showed non-significant difference between treatment and control groups. Present data suggest that liver and lung could serve as target organs in
oral toxicity of this plant.
2007 Elsevier Ireland Ltd. All rights reserved.
Keywords: Galega ofcinalis; Acute toxicity; Subchronic toxicity

1. Introduction
Galega ofcinalis (Papilionaceae) is a native plant from
southeastern Europe and was used as a treatment for diabetes in
medieval times (Oubre et al., 1997). The discovery of the plants
active hypoglycemic agent led to the development of metformin,
a biguanide which has been used to treat type 2 diabetes mellitus (Oubre et al., 1997; Palit et al., 1999). In fact, the only
example of an approved antidiabetic drug that was developed
from an herbal source with a long history of use for diabetes
is metformin from Galega ofcinalis (Vuksan and Sievenpiper,
2005).
Also in traditional medicine, Galega ofcinalis (syn.: galega)
is well known as a galactagoguing plant to improve milk
secretion, both in human and in animals (Uncini Manganelli
et al., 2001; Leporatti and Ivancheva, 2003). Recent experimental investigations indicate that crude aqueous extracts and
gel-filtered fractions of the plant suppress platelet aggregation
(Atanasov, 1994; Atanasov and Spasov, 2000). Moreover, it was

Corresponding author. Tel.: +98 21 88773521; fax: +98 21 88795008.


E-mail address: hrasekh@gmail.com (H.R. Rasekh).

0378-8741/$ see front matter 2007 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/j.jep.2007.10.030

found that active platelet disaggregating fraction of the extract


appeared to be a polysaccharideprotein complex (Atanasov and
Tchorbanov, 2002). Alcoholic extract of Galega ofcinalis was
tested on gram-positive and gram-negative bacteria as the plant
was claimed to hasten skin healing after surgery. This ethanolic
(60%) extract exhibited significant inhibition on growth of both
gram-positive and gram-negative bacteria (Pundarikakshudu et
al., 2001). In addition to above established effects, the plant has
weight reducing action, the mechanism of which is unclear but
involves loss of body fat (Palit et al., 1999).
Previous studies found the plant Galega ofcinalis to be
toxic to sheep and are capable of causing death. The animals
usually presented with hydrothorax, accumulation of a frothy
fibrinous fluid in airways, and pulmonary congestion and edema
(Puyt et al., 1981; Keeler et al., 1986, 1988). In some cases,
subendocardial hemorrhage was present and the most consistent
microscopic changes were found in the lungs with congestion of
alveolar capillaries and eosinophilic protein-rich edema fluid. It
is believed that galegine is the major compound in the plant that
causes injury (Keeler et al., 1992).
Considering both the ethnobotanical and pharmacological
applications of the plant, the aim of this study was to investigate the possible toxic effects of the aerial parts of plant in rats

22

H.R. Rasekh et al. / Journal of Ethnopharmacology 116 (2008) 2126

using histopathological and hematological examinations as well


as biochemical parameters.
2. Materials and methods
2.1. Plant material
Galega ofcinalis has been harvested in flowering period by
a pharmaceutical company (Zard Band Co., Tehran, Iran). The
company has supplied the dried aerial part of plant in powdered form. The plant was authenticated by M. Kamalinejad,
Department of Pharmacognosy and a voucher specimen (Registration Number 94) has been deposited in the Herbarium of
the School of Pharmacy (Shaheed Beheshti Medical University,
Tehran, Iran).
2.2. Evaluation of toxicity following single dose
administration
2.2.1. Animals
Four to 5 weeks old, Wistar rats of both sexes were purchased
from Razi Research Institute (Hesarak, Karaj, Iran) and acclimated to holding facilities for 2 weeks prior to dosing. Animals
were randomly assigned to control and four treatment groups
(5 rats/(sex group)) and were housed in clear plastic cages containing wood shavings for bedding. Each cage contained five
rats of the same sex and were fed on normal laboratory chow
(Pars Co., Tehran, Iran) and given tap water ad libitum throughout the study. Environmental conditions were maintained at a
temperature of 23 2 C and a relative humidity of 40 10%
with 12 h light/dark cycle. At the onset of dosing, males weighed
172 23 g and females weighed 135 11 g. The research was
conducted in accordance with the internationally accepted principles for laboratory animals use and care as found in the US
guidelines (NIH Publication no. 8523, revised in 1985).
2.2.2. Administration
Animals were fasted for 12 h prior to dosing on day 0.
Treatment rats were dosed by oral gavage, using a curved, balltipped stainless steel feeding needle, with aqueous suspensions
of ground very fine powder of Galega ofcinalis. The dosages
delivered were 0.5, 1, 2.5, and 5 g/kg body weight galega herb
in the constant volume of 10 ml/kg body weight. The controls
received tap water by gavage in the same volume.
2.2.3. Observations
All rats were monitored continuously for 10 h after dosing for
signs of toxicity. For the remainder of the 14 days study period,
animals were monitored daily for mortality and any changes in
food and water consumption, and any additional behavioral or
clinical signs of toxicity. Animals body weight were measured
prior to dosing and on days 7 and 14. On day 14, all animals were
killed and at the end of the study the number of dead animals
was expressed in percentage and, if possible, the LD50 value
was established using Probits method (Angelis Pereira et al.,
2003).

2.3. Evaluation of toxicity following subchronic treatment


2.3.1. Animals
Four weeks old Wistar rats of both sexes were obtained from
Razi Research Institute and acclimated for 4 weeks prior to the
start of study. They were randomly divided into control and
three treatment groups (6 rats/(sex group)). Animals were caged
in pairs of the same sex in clear plastic cages containing wood
shavings for bedding. At the time when dosing was initiated, rats
were 8 weeks old, and males weighed 190 14 g and females
weighed 153 16 g.
2.3.2. Treatment
For the purposes of this study, the ground plant material
was incorporated into the standard laboratory rat diet (Pars
Co., Tehran) at constant concentrations of 0.1%, 1% and 2%
(w/w), by mixing with powdered standard diet. Our pilot study
indicated that based on daily food consumption of animals,
these concentration would achieve daily doses of 0.1, 1, and
2 g/kg body weight, respectively. Finally, the galega containing diet was repelleted before being supplied to rat. Control
diet was prepared similarly but without the addition of plant
material.
The prepared pellets were given to rats as their daily diet
(instead of standard diet) for 13 weeks, to determine the adverse
effects of Galega ofcinalis.
Six weeks after the onset of treatment, due to a reduction
in food consumption by animals, the concentration of plant
material in diet was increased to reach 0.15%, 1.5% and 3%
(w/w) of the prepared diet (Keyler et al., 2002). The animals
had free access to food and tap water ad libitum throughout the
test.
2.3.3. In life evaluations
Observations of mortality and toxicological signs were made
daily for 13 weeks. The time of onset, intensity, and duration of
these symptoms, if any, was recorded. The weight of each rat
was recorded on day 0 and at weekly intervals throughout the
course of the study. Food and water consumption were measured
three times a week.
2.3.4. Biochemical and hematological analyses
At the end of the study, all animals were fasted for 12 h and
then anaesthetized with an ip injection of a mixture containing
ketamine (40 mg/kg) and xylazine (10 mg/kg). The jugular vein
was exposed, and blood samples were taken by jugular vein
puncture (Wilson et al., 2001).
Blood samples for biochemical analyses were centrifuged at
3000 g for 5 min and the plasma collected and analyzed for
glucose, blood urea nitrogen, creatinine, total protein, albumin,
albumin/globulin ratio, phosphorus, calcium, sodium, potassium, chloride, bilirubin (total and conjugated), cholesterol,
triglycerides, alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate
dehydrogenase (LDH) and creatin phosphokinase (CPK). These
levels were determined by COBAS Mira S chemistry analyzer
(Roche Diagnostic Systems).

H.R. Rasekh et al. / Journal of Ethnopharmacology 116 (2008) 2126

23

Hematological analyses were done using standard techniques. Erythrocyte (RBC), total and differential leukocyte
(WBC), hematocrit (Hct), hemoglobin (Hb), platelet count,
mean corpuscular volume (MCV), mean corpuscular
hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), mean platelet volume (MPV), platelet
distribution width (PDW), and red distribution width (RDW)
of blood samples were determined using Sysmex K-1000 fully
automated hematology analyzer.
2.3.5. Necropsy
Following blood collection, rats were sacrificed by decapitation and liver, heart, lung, and kidney of all animals and ovary in
female rats were removed. The macroscopic appearance of the
organs was noted and their weights were recorded. Livers and
lungs of the half of animals in each group (3 rats/(sex group))
were preserved in 10% neutral buffered formalin for microscopic
histopathological examinations. The tissues were embedded in
paraffin, and then sectioned, stained with haematoxylin and
eosin and were examined microscopically.
2.3.6. Statistical analysis
Analysis of one-way variance (ANOVA) and Dunnett test
was used to determine statistical significance between tests and
controls. Only the values with a p < 0.05 were considered statistically significant (Yamakoshi et al., 2002; Bidhe and Ghosh,
2004).
3. Results
3.1. Acute study
No mortalities had occurred during the study and clinical
observations and measurements did not indicate evidences of
substance-related toxicity. On gross histopathological observation, rare red spots indicating lung hemorrhage in some of
animals in the highest dose group of both sexes were found. The
oral acute toxicity of Galega ofcinalis was, therefore, considered as unclassified, since a dose of 5 g/kg did not induce deaths
or toxic symptoms.
3.2. Subchronic study

Fig. 1. Changes in female rats body weight with duration of treatment. Each
point represents mean S.D. of N = 6.

PDW levels for control rats were not significantly different from
those in treatment groups. However, WBC counts and MPV were
decreased significantly (p < 0.05) in 1.5% group of male rats.
A statistically significant decrease of platelet count in 0.15%
(p < 0.05) and 1.5% (p < 0.01) groups was also observed. In
females, a reduction in hematocrit levels in 1.5% group (p < 0.05)
had occurred.
3.2.3. Biochemical analysis
A slight, but significant reduction in urea was noted in
high dose male rats (Table 2). Cholesterol levels were elevated significantly among middle and high dose groups of
both sexes (p < 0.01). Calcium ion concentrations were significantly reduced in all treatment groups of females (p < 0.01). This
parameter in males had a reduction in the 0.15% (p < 0.05) and
elevation in the 3% (p < 0.01) groups. Phosphorous concentrations in the 0.15% (p < 0.01) and 3% (p < 0.05) groups of males,
albumin levels in the middle dose group (p < 0.05) and albumin/globulin ratio in middle and high dose groups (p < 0.01)
of females were decreased significantly. The mean LDH levels in the middle (p < 0.01) and high dose (p < 0.05) females
were significantly increased whereas, this parameter shows a
reduction in the 3% group (p < 0.01) in males. There was a statistically significant increase in CPK level in all female treatment
groups (p < 0.01). Total bilirubin in 1.5% females and conjugated bilirubin in 1.5% males and 3% females were increased
(p < 0.05).

3.2.1. Body weight and food and water consumption


There was no significant difference in body weights between
control and treatment groups (Figs. 1 and 2). There was no
significant difference between food and water consumption of
galega treated animals compared with control. The intakes of
Galega ofcinalis at the 0.15%, 1.5%, and 3% dietary levels were approximately equal to intakes of 89.0 14.2 mg,
0.91 0.13 g, and 1.84 0.24 g/kg body weight in females
and 92.9 13.2 mg, 0.94 0.17 g, and 1.89 0.31 g/kg body
weight in males, respectively.
3.2.2. Hematological analysis
Results of the hematological studies are presented in Table 1.
The data show that RBC, Hb, MCV, MCH, MCHC, RDW, and

Fig. 2. Changes in male rats body weight with duration of treatment. Each point
represents mean S.D. of N = 6.

24

H.R. Rasekh et al. / Journal of Ethnopharmacology 116 (2008) 2126

Table 1
Hematological parameters at termination of treatment
Sex

Dose (%)

WBC (103 /l)

Male

Control
0.15
1.5
3

9.25
5.02
3.30
7.83

1.79
1.03
1.34*
0.80

8.50
7.94
6.99
8.15

0.09
0.17
1.18
0.22

15.47
14.38
12.12
15.15

0.16
0.58
2.86
0.26

47.17
43.52
38.00
44.60

0.60
1.03
6.70
0.84

55.67
54.83
54.00
55.00

0.56
0.70
0.71
1.73

Female

Control
0.15
1.5
3

5.37
4.87
4.55
5.73

0.65
0.83
0.69
0.63

7.72
7.54
6.54
7.35

0.14
0.17
0.88
0.14

14.30
13.95
11.97
13.80

0.22
0.38
1.49
0.65

43.40
40.15
34.7
40.35

0.60
0.95
3.96*
1.17

56.17
53.33
54.83
54.63

0.54
0.67
2.70
0.86

MCH (pg)
18.17
18.00
16.25
18.50
18.50
18.67
18.83
18.67

0.17
0.45
2.10
0.50
0.34
0.21
0.91
0.56

MCHC (%)
32.83
33.17
30.00
34.00
32.83
34.83
34.33
34.00

0.17
0.91
3.34
0.26
0.40
0.48
1.02
0.86

RBC (106 /l)

MPV (fl)
8.28
8.30
7.65
8.22
7.55
7.68
7.80
7.53

0.10
0.10
0.06*
0.22
0.08
0.09
0.12
0.11

Hb (g/dl)

HCT (%)

Platelets (105 /l)


9.10
4.87
3.13
9.19
11.05
9.52
8.15
9.52

1.00
0.88*
1.08**
1.03
0.33
0.61
1.05
1.38

MCV (fl)

RDW (%)
16.62
15.47
16.52
16.37
15.48
15.50
14.62
14.73

0.21
0.53
0.26
0.26
0.41
0.07
0.35
0.34

PDW (%)
9.90
10.93
10.17
9.62
8.68
9.03
9.03
8.95

0.15
0.78
0.51
0.38
0.09
0.23
0.16
0.23

Data presented as mean S.E.M. for N = 6. Significantly different from control: *p < 0.05, **p < 0.01.

3.2.4. Organ weights and histopathology


Relative organ weights measurements data are presented
in Table 3. There were no remarkable differences in relative
weights of kidney, lung, heart, and ovary. An increase in relative liver weight was observed in the middle and high dose males
(p < 0.01) and high dose females (p < 0.05). This elevation ranges
to 1520% above that of controls.
During necropsy, one male rat of low dose group had lung
hemorrhage, but no gross lesions were found in the rest of animals. The results of microscopic examination of control and
treatment animals show intraalveolar hemorrhage, intraalveolar
neutrophils, and alveolar edema in lung and sinusoidal congestion and kupffer cells hypertrophy in liver. These lesions
were observed in treatments but not control animals. However,
since incidence and severity of intraalveolar neutrophils, alveolar edema, and Kupffer cells hypertrophy is rare and mild,
they are not toxicologically significant. Determinations of other
parameters including alveolar collaps, septal thickness, interstitial infiltrate, and congestion of alveolar capillaries in lung and
fatty changes, hepatocyte regeneration/degeneration, architectural distortion, necrosis, interface hepatitis, portal infiltration,
and obstruction/dilatation of the bile duct in liver demonstrated
no treatment related lesions in organs examined.
4. Discussion and conclusions
The oral LD50 value in this study suggest that Galega ofcinalis is a relatively nontoxic plant.
The feeding of galega at dietary levels up to 3% to male
and female Wistar rats for 90 days was not associated with
any mortalities and abnormalities in general conditions, behavior, growth, and food and water consumption of animals. Body
weight gain and/or food consumption levels were similar in
both control and treated animals. The weight reducing effect
of galega in mice had been investigated previously. It was estab-

lished that, this plant has a weight reducing action in both normal
and genetically obese mice (Palit et al., 1999).
In an earlier toxicity study in sheep, no significant alterations in blood constituents were observed in treated animals
(Keeler et al., 1986). However, in present study a reduction in
Hct in females and WBC, platelet counts and MPV in males
were seen. The reduction of Hct is a result of slight and nonsignificant reduction in RBC, WBC, and platelets in females
and it does not appear to be toxicologically significant. It seems
that galega has toxic effects on bone marrow which leads to
reduce platelets count and volume and WBC count. Although,
it is not an established conclusion and surely needs more
investigations.
The results of the present study indicate that significant alterations in liver function have occurred. Urea reduction is a result
of liver damage. Hypercholesterolemia, another significant incident, could be an indicator of hepatic function failure. Although,
usually liver damage occurs along with fat accumulation in hepatocytes, this study did not show any remarkable pathological
changes including fatty changes in liver. Moreover, some other
parameters like bilirubin elevation and albumin reduction are
confirmations to varying liver function. A small elevation in
plasma bilirubin is an important indicator of liver damage in
laboratory animals or could be a sign of biliary duct obstruction. In liver microscopic examination, there is no obstruction
in intrahepatic bile duct, so it could be a result of problems
in extrahepatic bilirubin pathway. If, this problem continues,
hypercholesterolemia and ALP increase will be developed. It
should be noted that in this study any elevations in ALP or other
liver enzymes did not occur.
Determination of plasma proteins like albumin can act as a
criterion for assessing synthetic capacity of the liver, since nearly
all of them synthesized in hepatocytes. Decrease in plasma proteins therefore tend to reflect chronic damage. The common
pattern seen following significant hepatocellular damage is a

H.R. Rasekh et al. / Journal of Ethnopharmacology 116 (2008) 2126

25

Table 2
Biochemical parameters at termination of treatment
Sex

Dose (%)

FBS (mg/dl)

Male

Control
0.15
1.5
3

124.2
179.2
177.2
163.7

12.7
9.1
31.7
14.7

0.72
0.63
0.73
0.72

0.04
0.02
0.03
0.02

47.17
42.17
40.75
39.00

1.92
1.97
1.65
1.79*

48.83
53.17
55.25
58.33

4.12
3.66
11.83
10.63

Female

Control
0.15
1.5
3

146.7
130.3
132.0
111.3

11.9
10.0
10.9
8.1

0.75
0.65
0.73
0.60

0.08
0.08
0.12
0.09

50.50
48.33
42.67
48.67

3.75
3.36
5.43
4.89

65.00
84.50
36.00
74.00

13.58
14.81
4.13
17.09

Cholesterol (mg/dl)
68.33
76.00
89.25
84.50
106.50
128.17
145.33
141.33

Na (mEq/L)

2.03
2.71
4.70**
3.95**
1.73
11.60
5.23**
4.69**

140.0
138.0
138.5
144.5
140.3
136.3
138.0
137.7

Creatinine (mg/dl)

K (mEq/L)

0.63
0.45
1.32
4.17
1.23
1.45
1.46
0.92

5.48
5.42
6.07
5.42
7.77
7.55
7.22
7.43

Urea (mg/dl)

Cl (mEq/L)

0.27
0.14
0.39
0.29
0.07
0.16
0.20
0.11

94.83
94.50
99.75
90.33
95.67
89.33
91.50
96.17

Ca (mg/dl)

4.33
3.84
5.76
3.61
4.22
2.72
3.81
2.89

8.53
6.88
8.90
10.43
10.57
5.35
3.35
3.25

0.31
0.11*
0.92
0.36**
0.31
1.17**
0.26**
0.15**

11.10
8.07
9.47
8.57
11.92
13.82
11.93
12.00

0.81
0.47**
0.52
0.53*
0.94
1.37
0.85
0.92

Total protein (mg/dl)

Male

Control
0.15
1.5
3

8.53
7.97
8.47
8.30

0.24
0.16
0.22
0.21

4.52
4.35
4.37
4.17

0.15
0.10
0.09
0.10

1.12
1.22
1.05
1.02

0.08
0.10
0.05
0.03

219.0
212.7
181.3
183.7

17.6
23.8
6.3
24.4

66.00
65.33
52.33
65.83

4.19
7.61
6.74
6.34

Female

Control
0.15
1.5
3

8.73
9.05
8.93
9.53

0.29
0.54
0.17
0.55

4.67
4.57
3.40
3.80

0.26
0.33
0.17*
0.38

1.17
1.05
0.62
0.65

0.11
0.10
0.06**
0.06**

204.5
220.2
224.3
221.0

1.4
28.0
10.1
23.2

65.50
63.83
42.67
62.67

12.65
9.50
1.98
10.52

2209
1965
2150
1033
1713
2702
3392
3013

109
107
72
324**
325
355
430**
210*

1212
1092
1261
864
954
2277
2771
2465

ALP (IU/L)

28
146
97
237
185
350**
103**
167**

289.3
250.8
317.7
267.2
153.5
250.4
114.0
252.7

AST (IU/L)

Dose (%)

Total CPK (U/I)

Al/G ratio

Phosphorous (mg/dl)

Sex

LDH (U/I)

Albumin (gr/dl)

Triglycerides (mg/dl)

T. bilirubin (mg/dl)

28.0
44.1
45.7
46.3
65.9
49.9
35.8
37.7

0.35
0.33
0.45
0.48
0.58
0.72
1.13
0.93

ALT (IU/L)

Conj. bilirubin (mg/dl)

0.15
0.03
0.17
0.08
0.08
0.13
0.12*
0.22

0.08
0.10
0.15
0.10
0.12
0.35
0.35
0.55

0.02
0.00
0.03*
0.00
0.02
0.09
0.05
0.22*

FBS = fasting blood sugar, Na = sodium ion, K = potassium ion, Cl = chloride ion, Ca = calcium ion. Al/G = albumin/globulin Ratio, T. = total, Conj. = conjugated.
Data presented as mean S.E.M. for N = 6. Significantly different from control: *p < 0.05, **p < 0.01.

reduction in albumin accompanied by a relative increase in globulins which leads to albumin/globulin ratio reduction, often with
little change in the level of total protein (Woodman, 1996). This
pattern was seen in the present study, although microscopic

examination showed normal hepatocytes without any lesions


in liver. Therefore, it is suggested that galega causes liver function changes which cause fluctuation in above parameters but
the types and mechanisms of changes are unclear. More investi-

Table 3
Relative organ weight at termination of treatment (g % body weight)
Sex

Dose (%)

Liver%

Male

Control
0.15
1.5
3
Control
0.15
1.5
3

2.69
2.79
3.09
3.13
3.13
3.01
3.13
3.77

Female

Kidney%
0.03
0.07
0.11**
0.07**
0.09
0.18
0.11
0.27*

0.29
0.32
0.35
0.35
0.35
0.36
0.35
0.36

0.01
0.01
0.04
0.01
0.01
0.01
0.01
0.02

Heart%
0.32
0.29
0.35
0.35
0.37
0.38
0.36
0.37

Data presented as mean S.E.M. for N = 6. Significantly different from control: *p < 0.05, **p < 0.01.

Lung%
0.01
0.01
0.02
0.01
0.01
0.01
0.01
0.02

0.50
0.64
0.50
0.52
0.70
0.66
0.72
0.70

Ovary%
0.02
0.08
0.03
0.02
0.04
0.02
0.03
0.02

0.07
0.05
0.06
0.06

0.00
0.00
0.01
0.01

26

H.R. Rasekh et al. / Journal of Ethnopharmacology 116 (2008) 2126

gations on liver metabolic enzymes and/or electron micrograph


determinations of liver are needed.
Another significant factor is galega-induced hypocalcemia in
females. Albumin binds to calcium and reduced plasma albumin
levels lead to reduced levels of this cation in plasma. Remarkable
elevation in CPK and LDH, initial indicators of myopathy, in
females represents potentials for muscular damage. These two
enzymes have diagnostic value in myocardial or skeletal muscle
injuries. Hence, it could be suggested that Galega ofcinalis may
cause muscular damage in females.
A dose-dependent increase in relative liver weight observed
in animals could also be significant. Sinusoidal congestion by
reserving blood in liver may cause the relative liver weight elevation.
A compared analysis of lung microscopic findings from the
present study with those performed previously indicates similar toxicosis in respiratory system. Puyt (Puyt et al., 1981)
have reported some cases of poisonings due to galega herb
ingestion in sheep. The poisoning was characterized by an
asphyxic syndrome which leads to death within hours. Moreover, other studies confirmed fatal poisoning accompanied by
severe pulmonary problems (Keeler et al., 1986, 1988, 1992). In
another experiment (Lopez et al., 1996), toxicity of Verbesina
encelioides which has the same active substance as Galega
ofcinalis, galegine, was evaluated. Microscopically, severe
glomerulonephrosis and congestion in the liver, with cellular
degeneration and fatty changes were observed in sheep received
6.3 g/kg body weight of dried plant material. Additionally, hemorrhagic lymph nodes, and hemorrhagic and edematous lungs
were noted.
In conclusion, a number of significant clinical and pathological changes were associated with the subchronic oral
administration of Galega ofcinalis to Wistar rats. Several of
these observations support the conclusion that lung and liver
are plausible target organs for both sexes, although the mechanism of toxic effect on liver is unclear. Based on the results of
this study it can be concluded that the risk of oral toxicity to
mammals is not negligible.
Acknowledgements
The authors would like to express their appreciation to Dr.
Sarang Younesi, for his initial assistance in blood analysis and
Dr. Bahram Shahgoli, for examining the histological slides.
The authors are grateful to Majid Aslani and Farahnaz Tanbakousazan, for their technical assistance.
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