Hepatoprotective
Activity of Aqueous and Alcoholic Extracts from Corm’s of Amorphophallus paeonifolius
Against Carbontetra Chloride Induced Hepatotoxicity in Albino Rats
K Kathiresan*, S Tom, VV Venkatachalam and
H Penchalaiah
Department of Pharmacy,
ABSTRACT:
In the present study, the pre-treatment of
rats with Amorphophallus paeoniifolius. alcoholic and aqueous extracts
protected the animals against CCl4 induced hepatotoxicity. the treatment significantly reduced the
serum GOT, GPT, ALP, oxidation of GSH, GST levels and liver weight. The
hepatoprotective nature of aqueous and alcoholic extracts of this plant against
CCl4 induced hepatic oxidative stress may be attributed to the
presence of phenolic compounds. Further this was evidenced by free radical
scavenging activity, reduced lipid peroxidation, reduced glutathione (GSH)
radical scavenging activity and nitric oxide scavenging activity by both
extracts of Amorphophallus paeonifolius. The present study reveals that Amorphophallus
paeoniifolius possesses a hepatoprotective action.
KEYWORDS: Amorphophallus
paeonifolius, Aqueous and
Alcoholic extracts, hepatoprotective, hepatotoxity.
INTRODUCTION:
The liver is an organ of paramount
importance. It plays a pivotal role in the metabolism of xenobiotics. Herbal
medicines1-4 have received great attention as an alternative to
conventional therapy and the demand for these remedies has currently increased
(De Smet, 2002). Consumption of herbs and vegetables are known to benefit life
via the prevention of life style-related diseases including liver disorders.
This present study is on Aqueous and Alcoholic extract from corms of
amrophallus paeoniifolius to prevent against carbon tetra chloride induced
hepatotoxicity5-9 in Albino rats.
The dried corm of this plant was collected
and coarsely powdered. The powder was then successively extracted with ethanol
and distilled water using soxhlet
extractor. The aqueous and alcoholic extracts5 were dried under
reduced pressure using a rotary flash evaporator and they were kept under the
refrigeration till conducting further studies. The extracts thus obtained were
used for the preliminary photochemical screening and pharmacological studies10-20.
The extracts were administered to the
animals as suspension in 1 % aqueous gum acacia. The percentage yields of
aqueous and alcoholic extracts were 7 % and 6% respectively.
Wistar albino rats of either sex of 8 – 10
weeks old, weighing 180-200 gm, obtained from the experimental animal care
center, Central Animal house, (Reg No: 160/1999, CPCSEA, Proposal No:236,
Approved Date: 04.10.2004) college
of pharmacy, Annamalai
University, chidambaram, were
used.
The
animals21, 22, 23 were kept at a constant temperature (22±2°C),
humidity (55%) light-dark conditions (12/12 h light / dark ratio). The animals
were provided with purina chow and free access to drinking water ad libitum.
The conduct of experiments and the procedures of sacrifice were approved by the
Ethics Committee of the Experimental animal care society, college of pharmacy, Annamalai University, Chidambaram, India.
Phytochemical Screening
A phytochemical24, 25 analysis of the
color ofAmorphophallus Paeoniifolius was conducted for the detection of
alkaloids, cardiac glycsides, flavounoids, tannins,
anthraquinones, saponinns, volatile oil, cyanogenic glycosides,
coumarins, sterols, triterpenes and sulphur containing compounds (Farns Worth,
1966).
Study Protocol:
The animals were divided into ‘7’ groups consisting of
‘six’ rats in each group.
|
Group 1: |
Animals were received single daily dose of 1% aqueous
acacia on all 5 days (1 ml / kg P.O) and olive oil (1 ml / kg, S.C.) on days
2 and 3. |
|
Group 2: |
Animals were received single daily dose of 1% aqueous
acacia (1 ml / kg, P.O.) for 5 days. |
|
Group 3: |
Animals were treated with 200 mg / kg P.O. of aqueous
extract of Amorphophallus paeoniifolius corm on all 5 days. |
|
Group 4: |
Animals were treated with 400 mg / kg, P.I. of
aqueous extract of Amorphophallus paeoniifolius corm on all 5 days. |
|
Group 5: |
Animals were treated with 200 mg / kg P.O. of
alcoholic extract of this plant on all 5 days. |
|
Group 6: |
Animals were treated with 400 mg / kg P.O. of
alcoholic extract of this plant on all 5 days. |
|
Group 7: |
Animals were treated with 1 ml / kg P.O. of liv-52
syrup on all 5 days. |
For all groups (except group 1) commonly administered
carbon tetrachloride (2 ml / kg, S.C.) on day 2 and 3, 30 minutes after
administration of respective extract.
the serum levels of glutamic oxaloacetic transaminase (GOT)26,
glutamic–pyruvate transaminase (GPT)26,alkaline Phosphatase (ALP)27,
reduced Glutathione (GSH)28- 30, liver weight and
glutathione-stransferase (GST)31.
Group I: Dilated central vein,
normal hepatocytes.
Histopathological Study of
Group II: Perivenular inflammatory infiltration and
hepatocytic fatty change, diffuse mild hepatocellular vacuolation
Histopathological Study of
CCl4 toxic animal (H and E X100)
Aqueous and alcoholic extracts of Amorphophallus
paeoniifolius corm inhibited ferrous sulphate induced lipid per oxidation
in a dose dependent manner. The IC50 values of aqueous and alcoholic
extract were found to be 539.82 mg/ml (r=0.90) and 527.73 mg/ml (r =
0.90).
Group III :Change central
vein, mild fatty change.
Histopathological Study of
Aqueous extract 200 mg/kg and CCl4 treated animal (H and E X100)
Group IV: Dilated central vein, mild sinusoidal
dilation - No hepatocellular damage. (Near normal)
Histopathological Study of
Aqueous extract 400 mg/kg and CCl4 treated animal (Group IV) (H and
E X100)
Group V: Perilobular hepatocellular fatty change, (mild
fatty change), Peripherral lobule Histopathological
Study of Alcoholic extract 200 mg/kg and CCl4 treated animal (H and
E X100)
|
S. No |
Parameter |
GROUP |
Significance |
|||||||
|
I |
II |
III |
IV |
V |
VI |
VII
|
|
|||
|
1. |
Body weight
(mg/g) |
Liver weight |
33.15 ±0.671 |
46.49 ±0.376a |
41.53 ±0.442b |
36.98 ±0.461b |
38.17 ±0.497b |
37.24± 0.519b |
35.41± 0.269b |
P value – a < 0.001 Vs group I, b
< 0.001 Vs group II, values are mean ± SE of 6 animals in each group |
|
2. |
Glutamate oxaloactetate transaminase (GOT/AST) |
Serum (U/ml) |
28.83 ±0.87 |
179.16 ±0.364a |
122.83 ±2.23b |
34.00 ±1.53b |
123.83 ±3.66b |
43.20± 3.66b |
31.33 ±1.28b |
P value – a < 0.001 Vs group I, b
< 0.001 Vs group II, values are mean ± SE of 6 animals in each group |
|
Liver (mmol of pyruvate liberated/mg protein /min) |
30.66 ±0.76 |
171.83 ±0.96a |
107.33 ±2.82b |
42.20 ±3.39b |
107.83 ±3.18b |
37.66 ±1.45b |
34.00 ±1.29b |
|||
|
3. |
Glutamate pyruvate
transaminase (GPT/ALT) |
Serum (U/ml) |
16.66 ±0.61 |
124.33 ±216a |
74.33 ±2.66b |
24.83 ±1.28b |
65.66 ±2.90b |
22.33 ±0.80b |
19.66 ±1.20b |
P value – a < 0.001 Vs group I, b
< 0.001 Vs group II, values are mean ± SE of 6 animals in each group |
|
Liver Umol of pyruvate liberated/mg protein/min |
14.83 ±0.40 |
99.33 ±1.12a |
67.5± 1.12b |
27.83 ±0.71b |
73.33 ±2.16b |
19.16 ±0.98b |
18.5 ±1.18b |
|||
|
4. |
Alkaline phosphatase (ALP) |
Serum |
115.43 ±1.66 |
234.66 ±5.35a |
208.21 ±3.01b |
157.42 ±3.01b |
2.2.54 ±4.92b |
160.72 ±1.43b |
146.74 ±2.36b |
P value – a < 0.001 Vs group I, b
< 0.001 Vs group II, values are mean ± SE of 6 animals in each group |
|
5. |
Alkaline phosphatase (ALP) |
0.30±0.01 |
1.919 ±0.12a |
0.832 ±0.04b |
0.832 ±0.04b |
0.624 ±0.01b |
0.820 ±0.10b |
.0597 ±0.10b |
0.493 ±0.01b |
P value – a < 0.001 Vs group I, b
< 0.001 Vs group II, values are mean ± SE of 6 animals in each group |
|
6. |
Total protein |
Serum (mg/dL) |
7.854±0.3 |
5.12±0.02a |
5.506±0.02b |
6.879±0.03b |
5.736±0.04b |
6.69±0.13b |
7.376±0.050b |
P value – a < 0.001 Vs group I, b
< 0.001 Vs group II, values are mean ± SE of 6 animals in each group |
|
Liver (mg/g tissue) |
0.7817 ±0.001 |
0.5164±0.004a |
0.5748±0.003b |
0.6529±0.022b |
0.5963±.0015b |
0.6963±0.004b |
0.7290±0.004b |
|||
|
7. |
Glutathione peroxide (GPx) (n moles of GSH oxidised/
min /mg protein) |
Liver |
314.11±0.06a |
190.85±4.941a |
2.38.44 ±10.063n |
270.33±5.069b |
214.38±7.89c |
2.6921±6.839b |
295.12±5.796b |
P value – a < 0.001 Vs group I, b
< 0.01 Vs group II, values are mean ± SE of 6 animals in each group |
|
8. |
Glutathione-S- transferase (GST) (n moles of CDNB
conjugate formed /min /mg protein) |
Liver |
0.334±0.001 |
0.223±0.008a |
0.256±0.001b |
0.298±0.001b |
0.251±.006c |
0.271±0.001b |
0.309±0.003b |
P value – a < 0.001 Vs group I, b
< 0.001 Vs group II, c < 0.01 Vs group V values are mean ± SE of 6
animals in each group |
|
9. |
Glutathione reductase (GRD) (n moles of GSSG utilized
/ mm/ mg protein) |
Liver |
25.56±0.348 |
14.38±0.335a |
16.16±0.187b |
18.54±0.120b |
15.99±0.353b |
18.29±0.239b |
20.36±0.295b |
P value – a < 0.001 Vs group I, b
< 0.01 Vs group II, values are mean ± SE of 6 animals in each group |
|
10. |
Superoxide dismutase (SOD) (Kat/g protein) |
Liver |
9.62± 0.261a |
6.79±0.229 |
7.50± 0.227b |
5.90± 0.483NS |
7.24± 0.289b |
8.56± 0.156b |
8.56± 0.156b |
P value – a < 0.001 Vs group I, b
< 0.001 Vs group II, c < 0.01 Vs group III, NS
Non-significant Vs group V values are
mean ± SE of 6 animals in each group |
|
11. |
Catalase (CAT) (n moles of H2O2
decomposed / mm/ mg protein) |
Liver |
77.51±0.257 |
52.22 ±0.272a |
60.88±0.579b |
69.69±0.410b |
60.71±1.545b |
69.37±0.452b |
74.29±0.389b |
P value – a < 0.001 Vs group I, b
< 0.01 Vs group II, values are mean ± SE of 6 animals in each group |
|
12. |
Lipid peroxidase (LPO) (n moles of nDA/mg protein) |
Liver |
3.51±0.029 |
1.65±0.042a |
1.89±0.023b |
2.19± 0.017b |
1.88± 0.021b |
2.29± 0.026b |
2.63± 0.017b |
P value – a < 0.001 Vs group I, b
< 0.01 Vs group II, values are mean ± SE of 6 animals in each group |
Group VI: Sinusoidal dilation
and peripheral hepatocytic fatty change. Histopathological
Study of Alcoholic extract 400 mg/kg and CCl4 treated animal (H and
E X100)
Aqueous and alcoholic extracts of Amorphophallus
paeoniifolius corm inhibited the oxidation of reduced glutathione in a dose
dependent manner. The IC50 values of aqueous and alcoholic extracts
were found to be 725.86 mg/ml (r=0.95) and 572.99 mg/ml (r=0.89).
CCl4 treated animals (Group II) showed a
significant increase (P<0.001) in wet weight of the liver compared to
control. There was a significant decrease in liver wet weight of animals
treated with 200 mg / kg and 400 mg / kg of aqueous and alcoholic extracts
(P<0.001), when compared with group II.
Glutamate oxaloacetate transminase (GOT) and
Glutamate pyruvate transaminase (GPT):
GOT and GPT levels of serum and liver homogenate were
significantly increased (P < 0.001) in group II animals challenged with CCl4,
when compared to control. A dose dependent reduction of (P < 0.001) GOT and
GPT levels were observed in animals treated with (200 mg / kg and 400 mg/kg) and alcoholic extracts (200 mg/kg and 400 mg/kg) when
compared to group II. Liv-52 (group VII) produced a significant reduction (P
< 0.001) at the dose of 1 ml / kg body weight / P.O. in both indices when
compared to group II.
GroupVII:
Histopathological study of Liv-52 syrup (1
ml/kg) &
CCl4 treated animal (H & E X100)
Serum Alkaline Phosphatase (ALP):
The serum ALP level was significantly increased (P <
0.001) in CCl4 challenged rats (group II) when compared to control
rats (group I). Treatment with Aqueous (200 mg/kg and 400 mg/kg) and alcoholic (200 mg/kg and 400 mg/kg) extracts
showed a significant (P < 0.001) reduction in ALP level when compared to
group II animals. The Liv-52 (1 ml / kg) treated group also showed a
significant decline of ALP (P < 0.001) when compared to group II animals.
Glutathione – S – Transferase (GST):
Liver Glutathione –S – transferase level was
significantly reduced (P < 0.001) in CCl4 treated animals, when
compared with normal animals. Both aqueous and alcoholic extracts of
Amorphophallus paeoniifolius corm at 200 mg/kg and 400 mg/kg dose levels showed significant increase
in GST level when compared to CCl4 treated group. Liv-52 1 mg/kg) treated
animals also showed significant increase of GST levels when compared to group
II.
GLOSSARY:
GOT : Glutamic oxaloacetic transaminase
GPT : Glutamic–pyruvate transaminase
ALP : Alkaline Phosphatase
GST : Glutathione –S – transferase
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Received on 19.01.2010
Accepted on 17.02.2010
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Research J. Pharmacology and
Pharmacodynamics 2(2): March –April 2010: 190-194