Effect
of Root Extracts and Isolated Compounds of Xanthium
strumarium L. on Ehrlich Ascites Carcinoma in Mice
N.B. Sridharamurthy1*,
R. Yogananda2 and Riyaz Mahamood3
1Department of Pharmacology, Dayanandsagar College
of Pharmacy, Banglore, Karnataka
India
2Department of Pharmaceutics, SJM college of
Pharmacy, Chitradurga, Karnataka, India,
3Chairman,
Department of Biotechnology, Kuvempu University, Shimoga, Karnataka, India,
ABSTRACT:
A mixture of Xanthine, Xanthone and Xanthosine isolated
from the root of Xanthium strumarium
L exhibited antitumor activities against Ehrlich Ascites Carcinoma in Swiss
albino mice. Animals were pre-treated orally with the extract at a dose of
200mg/kg, 25 and 50mg/kg body weight for isolated compounds of chloroform extract and then after 24hrs
EAC (at a dose of 2x106 cells/mouse) administration
and following an 18hr fasting; mice were sacrificed for studying of antitumor
activity. The decrement of tumor volume, packed cell volume and viable cell
count were observed in both lignans and isolated compounds of chloroform
extract treated animals and compared with only EAC tumor bearing mice.
Treatment with test compounds increased the survival time and normal peritoneal
cell count. Hematological parameters, PCV which were altered by tumor volume
inoculation, were restored considerably. Thus, this study was an attempt to
evaluate the preventive and curative role of X. strumarium lignans in
tumor bearing mice.
KEYWORDS: Ehrlich
Ascites Carcinoma, Xanthium strumarium,
lignans, xanthone, xanthine and xanthosine.
INTRODUCTION:
Cancer is the leading cause of mortality and most of
the chemotherapeutic agents have been reported to exhibit severe normal tissue
toxicity, accompanied by undesirable side effects. More over these drugs are
highly expensive mutagenic and carcinogenic. Therefore the substitute of the
conventional chemotherapeutic agents to control the high mortality rate are
needed which will be highly effective at non toxic doses and inexpensive and
accessible to general people1. This can be achieved by in-depth
research and continuous screening of new molecules or natural agents. The
natural medicines have played a great role to treat various disorders in humans
including cancer2, 3. Recent surveys indicate that plants,
vegetables and herbs as folk and traditional medicine have been accepted widely
as major resources of chemo preventive agents4. Xanthium
strumarium L. is an herb
commonly known as, rough cocklebur belonging to the family Asteraceae,
and found practically throughout India, Pakistan and Afghanistan. This plant
has been widely reported to have several medicinal properties in traditional
form of medicine.
The beneficial properties are diuretic, astringent,
sedative, demulcent, diaphoretic, analgesic, sialagogue, styptic,
sudorific, tonic, anodyne, antibacterial, antifungal, antispasmodic,
bactericide, bitter, depressant, hemostat, laxative, refrigerant,
antirheumatic, appetizer, emollient, bitter tonic, febrifuge and the treatment
of scrofulous tumors. There are several reports on the pharmacological
activities like anti-inflammatory, anti-tussive, anti-bacterial, nematosidal
and anti-fungal activities of the plant5. But there are no reports
on the anti-tumor properties. Therefore the present study was undertaken to
evaluate the anti-tumor properties for both root extracts and isolated
compounds of chloroform extract.
MATERIALS
AND METHODS:
Test compounds:
Collection and authentication
of plant material:
The roots of Xanthium strumarium L. were collected from uncultivated lands of Chitradurga
District, Karnataka state, India. The taxonomic authentication of the plant was
done by Prof. Sidalingappa, Head of the Dept of Botany, S J M Science College,
and Chitradurga, Karnataka state.
Three compounds xanthine, xanthosine and
xanthone were isolated from chloroform extract of the root and used for the
further investigation.
Preparation of extracts:
Various extracts of the plant
material were prepared by successive solvent extraction method as described
below.
The powdered material of roots of Xanthium
strumarium L was
refluxed successively with the solvents petroleum ether (400-600,
E- Merck Mumbai, India), Chloroform (500 – 700, E-Merck
Mumbai, India) and Ethanol (E- Merck Mumbai, India) in a soxhlet extractor for
48 hrs in batches of 350g each. Every time, before extracting with the next
solvent the marc was dried. After extraction with ethanol lastly, marc was kept
in closed jar in distilled water for 48 hrs with occasional shaking, then
distilled water obtained by pressing the marc with tincture press.
All the extracts were concentrated in vacuum using
rotary flash evaporator (Buchi-Flawil, Switzerland). The solvents were removed
completely over the water bath and finally desiccator dried. The extracts so
obtained from each of the solvents were labeled, weighed and the yield was
calculated in terms of grams percent of the weight of the powdered roots
Animals:
Healthy Albino
mice of either sex, weighing and 20-30 g
respectively were procured from the animal house of S J M. College of Pharmacy, Chitradurga, India. The animal
house was well ventilated and animals had 12 ± 1 h day and night schedule. The
animals were housed in large spacious hygienic cages during the course of the
experimental period and room temperature was maintained at 25 ± 1°C. The
animals were fed with standard rat feed (Hindustan Lever Ltd; Bangalore) and
water ad libitum. The experiments
were conducted as per the guidelines of CPCSEA, Chennai, India (approval no.
SJMCP /IAEC /PhD/ PH.CHEM /05/2007-08).
Tumor
cells and Inoculation:
Earlich ascites carcinoma (EAC) cells were obtained
through the courtesy of Amla Cancer Research Centre, Thrissur, Kerala state,
India. These cells were maintained in Swiss albino mice by weekly
intraperitoneal inoculation of 1x106 cells/mouse i.e., the viable
EAC cells were counted (Trypan blue indicator)
under microscope and were adjusted at 2x106cell/ml.
Preparation
of test samples:
Suspensions of the crude extracts viz; petroleum ether,
chloroform, ethanol and distilled water extracts were prepared in 0.5 % tween 80 so as to obtain the
dosage forms in the concentration of 20mg/ml. These suspensions were
administered orally to the animals with the help of intra-gastric catheter at
the dose 200 mg/kg of body weight for extracts.
Preparation
of Standard drug formulation:
The standard drug 5-Fluorouracil (5-FU) 100mg was
suspended in 100 ml of 0.5% tween 80 and administered orally to the animals
with the help of intra-gastric catheter at the dose 20 mg/kg of body weight.
Procedure
for Anti tumor testing:
The Swiss albino mice divided into seven
groups of eight animals each. All animals were inoculated with 0.2 ml of EAC cells per 10 g body weight of mouse on
the zero day expect Group I animals which are served as normal control received
only 0.5ml of 5% tween 80. A day of incubation was allowed for multiplication
of the cells. Fourteen dose of the ether, chloroform, ethanol, water extracts
and 5-Fluorouracil were given orally to the animals belongs group III to VII
with the help of intra-gastric catheter respectively from the first day up to
the 14th day with 24 h intervals. Group II animals served as tumor
control and received 0.5 ml of tween 80. Food and water withheld 18 h before
scarifying the animals. On 15th day, half of the animals (n=4) in
each group were anaesthetized slightly with anesthetic ether and blood was
collected from retro-orbital puncture and kept at 370C for 30 min.
Immediately
after collecting the blood samples, the mice were killed by cervical
dislocation. The blood collected was immediately used for the estimation of
hemoglobin (Hb) content, red blood cell count (RBC) and white blood cell (WBC).
The mean survival time (MST), percentage increase in life span (%ILS) and
percent increase in the body weight were calculated out from the rest of the
animals6.
Antitumor
activity of the extracts was measured in EAC animals with respect to the
following parameters:
i.
Tumour volume: The mice were dissected and the ascetic fluid was
collected from the peritoneal cavity. The volume was measured by taking it in a
graduated centrifuge tube and packed cell volume was determined by centrifuging
at 1000 rpm for 5 min.
ii.
Tumour cell count: The ascetic fluid was taken in a WBC
pipette and diluted 100 times. Then a drop of the diluted cell suspension was
placed on the Neubauer counting chamber and the number of cells in the 64 small
squares was counted.
iii. Viable / non-viable tumour
cell count: The
cells were then stained with trypan blue (0.4% in normal saline) dye. The cells
that did not take up the dye were viable and those that took the stain were
nonviable. These viable and nonviable cells were counted.
iv.
Percentage increase life span (% ILS): The effect of MEMP on
tumour growth was monitored by recording the mortality on daily basis for a period
of 6 weeks and percentage increase in life span7,8 27,28 (%ILS) was
calculated by using the formula;
v.
vi.
Body weight: Body weights of the experimental mice were recorded
both in the treated and control group at the beginning of the experiment (day
0) and sequentially on every alternative day during the treatment period
vii.
Heametological parameters: At the end of the experimental
period, all the mice were sacrificed after 18h of fasting by decapitation.
Blood was collected from freely flowing tail vein and used for the estimation
of Haemoglobin (Hb) content 930, red blood cells (RBC) 10,22count
and white blood cell (WBC)11,23 cell count. WBC differential count32
was carried out from Leishman stained blood smears12,24.
viii.
Estimation of Haemoglobin: 0.1ml of heparinised blood
was taken in Sahli’s hemoglobinometer and diluted with 0.1 NHcl until the colour matched with
standard. The reading was then taken from the graduated cylinder and expressed
as mg/100ml of blood.
ix.
Estimation of Erythrocyte
(RBC) count:
For the erythrocyte (RBC)
counts, blood samples were diluted 1:200 with the Rees-Ecker diluting fluid
containing Sodium citrate (3.8g), HCHO (0.2ml) and Brillint Cresyl Blue, volume
made up with 100ml of sterile distilled water.
Blood samples were drawn by
the fine tips (capacity20μl attached to an auto adjustable micropipette
transferred to the micro centrifuge tubes containing the dilution fluid. The
tubes were mixed thoroughly and a drop of the resultant suspension was
discharged under the cover glass of an improved Neubuer bright field
haemocytometer and the corpuscles were to settle. The number of erythrocyte in
80 small squares was counted under light microscope. The number of cells in
1μl of undiluted blood was calculated following the standard formula.
Erythrocyte count= N×
1/0.02×200.
N × 10,000. Where N = no.
of cells in 80 small squares i.e. 0.02μl
Short term toxicity studies:
For the determination of short term toxicity, healthy
Swiss albino mice were treated intra-peritonealy. The single doses of test
compounds viz., 100 to 200 mg/kg b.w of crude extracts and 25 and 50mg/kg b.w
of isolated compounds were administered to the experimental animals. The mice
which received 200mg/kg body weight of crude extracts and 50mg/kg body weight
of isolated compounds did not showed toxic effects. The symptoms were
inactiveness in general behavior, slow movement, loss of appetite, hypothermia,
erected hairs etc were absent. The weights of the vital organs (liver, kidney,
brain and spleen) were also not affected by these test compounds. Hematological
parameters like Hb, RBC and WBC, serum glutamate pyruvate transaminase (SGPT)
and serum glutamate oxaloacetate transaminase (SGOT) also remained unaltered.
RESULTS:
Among the four crude extracts of the root of X.
strumarium chloroform extract was found to be more potent against the EAC
tumor cell growth which can be indicated by the increase in mean survival time,
increase in lifespan and decrease in body weight 29.00±0.41, 31.81% and
15.42±0.78 respectively. When compare to EAC control 22.00±0.41 mean survival
time and 27.52±1.29 increase in body weight. Animals treated with known
standard drug 5-flurouracil showed maximum
increase in mean survival time 36.00±0.41, increase in lifespan up to 63.63% and decreased body weight
by whereas, the result of animals
treated with pet-ether, ethanol and aqueous extracts are shown in Table-1
Fig-1.
Table 1: Effect of Extracts of
Xanthium strumarium L. roots treatment on the Survival Time and Body
Weight of the tumor bearing mice.
|
Group No.
|
Mean survival time (MST) in days
|
Increase in life span ILS(%)
|
Increase in body weight IBW (%)
|
|
Normal control
EAC control (1x106cells)
EAC + Ether extract
EAC + Chloroform extract
EAC + Ethanol extract
EAC + Aqueous extract
EAC + 5FU(20mg/kgbw)
5- Fluorouracil standard drug
|
--
22.00±0.41
23.25±0.48
29.00±0.41
26.25±0.48
22.25±0.48
36.00±0.41
|
--
--
08.64%
31.81%
15.68%
03.68%
63.63%
|
--
27.52±1.29**
25.1 5±0.88**
15.4 2±0.78**
19.38±0.32**
27.15±1.49**
09.12±0.64**
|
Values are mean S.E.M. where
n=6; *p<0.001 statistically significant when compared with EAC control
group.
Fig-1: Effect of Extracts of Xanthium
strumarium L. roots treatment on the
Survival Time and Body Weight of the tumor bearing mice.
Mean survival time (MST) in
days
Increase in life span ILS(%)
Increase in body weight IBW
(%)
The tumor cell volume, packed cell volume and viable
cell count showed significant increase in EAC bearing mice control compared to
normal group. Administration of chloroform, extract showed a significant
reversal of these changes towards the normal values. The same effect was also
exhibited by the standard 5-fluorouracil treatment. The treatment with other
extracts showed less significant effect as shown in (Table No. 2, and Fig -2).
Fig-2: Effect of Extracts of Xanthium strumarium
L. roots treatment on the Survival Time and Body Weight of the tumor bearing
mice.
Tumor volume(mL)
Packed cell volume(mL)
Viable cells
Non-viable cells
The tumor cell volume, packed cell volume and viable
cell count showed significant increase in EAC bearing mice control compared to
normal group. Administration of xanthone
xanthosine and xanthine showed a significant (p<0.001) reversal of these changes towards the normal
values. The same effect was also exhibited by the standard 5-Fluorouracil
treatment. The treatment with other constituent showed less significant effect
as shown in (Table No. 3 and Fig-3).
Fig-3:
Effect of Isolated Constituents of
Xanthium strumarium L. roots treatment on the tumour growth.
Similar results were obtained among the animals treated
with isolated constituents of chloroform extract Xanthone and Xanthosine which
exhibited maximum protection against EAC bearing animals. It seems that Xanthone and Xanthosine protect
the mice against EAC tumor cells by decreasing the body weight, increase in the
lifespan and increase in mean survival time. The graphical representation and
the values are shown in Table-4, Fig-4.
Table-2: Effect of extracts of X. strumarium treatment on the tumour growth
|
Particulars Groups
|
Tumor volume(mL)
|
Packed cell volume(mL)
|
Viable cells
|
Non-viable cells
|
|
Normal control
EAC control (1x106cells)
EAC + Ether extract
EAC + Chloroform extract
EAC + Ethanol extract
EAC + Aqueous extract
EAC + 5FU(20mg/kg b.w) 5- Fluorouracil
standard drug
|
3.58±0.14
3.50±0.04**
1.75±0.04**
2.50±0.09**
4.10±0.05**
0.90±0.04**
|
2.45±0.17
2.25±0.06**
1.05±0.03**
1.51±0.03**
2.30±0.04**
0.55±0.03**
|
8.75±0.25
8.15±0.25**
4.00±0.41**
5.75±0.25**
8.0±0.4**
1.25±0.48**
|
0.75±0.25
0.50±0.29**
1.25±0.25**
1.75±0.25**
0.75±0.25**
0.75±0.25**
|
Values
are expressed as mean ± SEM, n=six rats in each group, ** p<0.001 compared
to tumour control
Table-3:
Effect of Isolated Constituents of
Xanthium strumarium L. roots treatment on the tumour growth.
|
Particulars
|
Tumor volume(mL)
|
Packed cell volume(mL)
|
Viable cells
|
Non-viable cells
|
|
Normal control
EAC control (1x106cells)
EAC + xanthone
EAC + xanthosine
EAC + xanthine
EAC + 5FU (20mg/kgbw) 5- Fluorouracil standard drug
|
3.85±0.14
1.20±0.11**
1.51±0.06**
2.51±0.06**
0.90±0.04**
|
2.35±0.17
0.75±0.12**
1.08±0.05**
2.00±0.04**
0.55±0.03**
|
8.25±0.25
3.50±0.29**
.00±0.41**
5.25±0.48**
1.25±0.48**
|
0.75±0.25
1.20±0.25**
1.15±0.25**
1.28±0.25**
0.75±0.25**
|
Values
are expressed as mean ± SEM, n=six rats in each group, ** p<0.001 compared
to tumour control
Table-4: Effect of Isolated
Constituents from chloroform extracts of Xanthium strumatrium L.
roots treatment on the Survival Time and Body Weight of the Tumor bearing mice.
|
GROUP NO
|
Mean survival time (MST) in days
|
Increase in life span ILS(%)
|
Increase in body weight IBW (%)
|
|
Normal control
EAC control (1x106cells)
EAC + Xanthone
EAC +Xanthosine
EAC + Xanthine
EAC + 5FU (20mg/kgbw)
5- Fluorouracil standard drug
|
--
22.00±0.41
33.31±0.25
34.40±0.34
24.25±0.24
36.00±0.41
|
--
--
51.40
56.36
15.68
63.64%
|
--
28.00±1.29
9.42±0.21**
9.35±1.39**
19.28±0.57**
9.12±0.64**
|
Values are mean S.E.M. where
n=6; *p<0.001 statistically significant when compared with EAC control
group.
Fig-4: Effect of
Isolated Constituents from chloroform extracts of Xanthium strumatrium
L. roots treatment on the Survival Time, Body Weight of the Tumor bearing mice.
Mean survival time (MST) in
days
Increase in life span ILS(%)
Increase in body weight IBW
(%)
The anti-tumor property of the four extracts and
isolated constituents were also assessed by studying the different
hematological parameters. Hematological parameters of EAC infected mice were
found to be remarkably altered compared to normal mice. The WBC count was
increased but the hemoglobin percentages which were found to be decreased with
modest change of the total number of RBCs. Treatment with chloroform extract
and its isolated constituents at the doses of 200mg/kg b. w and 25mg/kg b.w
respectively changed these altered parameters to the normal values. The altered
hematological parameters for both crude extracts and isolated constituents are
given in Tables-5, 6 and Figs-5, 6.
Table-5: Effect of Extracts of Xanthium strumarium
L. roots treatment on Haematological parameters of EAC bearing mice
|
Groups
|
Hb content
(g/dl)
|
Total RBC
count
106cells/mm3
|
Total WBC
count
103cells/mm3
|
Lymphocytes
(%)
|
Neutrophils
(%)
|
Monocytes (%)
|
|
Normal control
EAC control (1x106cells)
EAC + Ether
extract
EAC + Chloroform
extract
EAC + Ethanol
extract
EAC + Aqueous
extract
EAC + 5FU
(20mg/kgbw)
5- Fluorouracil
standard drug
|
13.15±0.10
09.20±0.11*
09.13±0.19**
12.58±0.16
10.10±0.19**
09.12±0.12**
13.12±0.05
|
6.00±0.09
3.85±0.14*
5.16±0.16
5.92±0.12
4.38±0.11**
3.90±0.15**
5.85±0.10
|
5.32±0.09
9.72±0.13*
9.25±0.16**
5.80±0.19
8.17±0.15**
8.80±0.11**
5.30±0.09
|
73.50±1.71
24.25±1.31*
41.75±1.21**
69.50±1.53
53.25±1.15**
49.25±1.55**
75.75±0.85
|
25.00±1.68
74.25±1.55*
68.15±1.08**
31.25±2.19**
72.00±1.18**
69.50±1.18**
23.00±0.82
|
1.50±0.29
1.40±0.29
1.48±0.29
1.35±0.25
1.45±0.25
1.52±0.29
1.25±0.25
|
Values are mean S.E.M. where n=6
*p< 0.001statistically significant when compared
with normal group;
**p < 0.001statistically significant when compared with
EAC control group.
Table-6 Effect of Isolated Constituents from chloroform
extracts of Xanthium strumarium L. roots treatment on Haematological
parameters of EAC bearing mice
|
GROUP NO
|
Hb content
(g/dl)
|
Total RBC
count
106cells/mm3
|
Total WBC
count
103cells/mm3
|
Lymphocytes
(%)
|
Neutrophils
(%)
|
Monocytes
(%)
|
|
1: Normal control
2: EAC control
(1x106cells)
3: EAC + Xanthone
4: EAC +
Xanthosine
5: EAC + Xanthine
6: EAC +
5FU(20mg/kgbw)
5- Fluorouracil
standard drug
|
13.15±0.10
09.20±0.11*
12.48±0.19**
12.54±0.29**
10.55±0.14*
13.12±0.05
|
6.00±0.09
3.85±0.14
5.49±0.18
5.11±0.13
4.47±0.17*
5.85±0.10
|
5.32±0.09
9.72±0.13*
5.13±0.05
5.19±0.16
6.10±0.12*
5.30±0.09
|
73.50±1.71
24.25±1.31*
72.32±0.45
64.15±1.42*
70.30±0.45*
75.75±0.85
|
25.00±1.68
74.25±1.55*
26.12±1.32
26.51±1.21*
34.00±0.31*
23.00±0.82
|
1.50±0.29
1.50±0.29
1.45±0.26
1.58±0.28
1.46±0.21
1.25±0.25
|
Values are mean S.E.M. where n=6
*p< 0.001statistically significant when compared
with normal group.
**p < 0.001statistically significant when compared
with EAC control group.
Fig-5: Effect of Extracts of Xanthium strumarium
L. roots treatment on Haematological parameters of EAC bearing mice
Fig-6: Effect of Isolated
Constituents from chloroform extracts of Xanthium strumarium L. roots
treatment on Haematological parameters of EAC bearing mice
In liver cell injury damage to the membrane of cells
and organelles allows intracellular enzymes to leak in to the blood. During
liver injury liver produces a large amount of aspartate amino transaminase
(AST) and alanine amino transaminase (ALT) which is secreted to the
circulation. Alanine amino transaminase (ALT) is a cytosol enzyme more abundant
in liver cells than any other cells in the body and is a more specific enzyme
for liver. Aspartate amino transaminase is a mitochondrial enzyme present in
large quantities in liver, heart, kidney and skeletal muscles. The elevated
activities of AST and ALT in the serum
indicates the possible occurrence of liver injury. The normal values of AST and
ALT ranges from 6-38 IU/L and 8-35 IU/L at 37° respectively. Administration of various extracts of the
root of X. strumarium and isolated constituents of chloroform extracts
alters the level of these enzymes and the level of bilirubin. Chloroform
extract and isolated constituents of chloroform extract exhibits a fall in the
level of AST ALT and bilirubin content Table- 7, 8. Fig-7, 8.
DISCUSSION
The present experiment was evaluated to investigate the
anti-tumor activity of the chloroform extract and isolated compounds of
chloroform extract of X. strumarium in EAC tumor infected
mice. The extract treatment of mice at the doses of 200mg/kg and 25mg/kg
pronouncedly inhibited the tumor volume, packed cell volume, total cell count
and also regained the hematological parameters near to normal condition.
Table-7: Effect
of Extracts of Xanthium strumarium L. roots treatment on marker enzymes
of EAC bearing mice.
|
Group (n)
|
ALT (IU/l)
|
AST (IU/l)
|
ALP (IU/l)
|
Total Bilirubin
|
|
Normal control
EAC control (1x106cells)
EAC + Ether
extract
EAC + Chloroform
extract
EAC + Ethanolic
extract
EAC + Aqueous
extract
EAC + 5FU
(20mg/kgbw) 5- Fluorouracil standard drug
|
181.5±5.06
292.00±8.75*
242.00±4.45**
228.75±7.47**
261.15±5.42**
267.72±3.43**
208.75±8.26*
|
63.42±3.44
202.62±9.30*
189.50±6.08**
114.50±4.95**
186.00±5.31**
194.32±3.56**
98.75±4.27*
|
221.50±5.45
271.50±6.30*
249.23±4.54**
202.31±9.21**
224.50±6.14**
263.5±4.13**
190.00±11.37*
|
0.56±0.02
4.88±0.10*
4.45±0.15**
1.25±0.17**
2.66±0.16**
4.45±0.19**
1.05±0.06*
|
|
ANOVA
f
df
p
|
35.8
6,21
0.0001
|
90.6
6,21
0.0001
|
19.1
6,21
0.0001
|
481.9
6,21
0.0001
|
*p values are expressed as mean ± SEM when n=6 rats in
each group.
*P< 0.001 compared to normal; **p < 0.001
compared to tumor control
Table-8: Effect of Isolated Constituents from
chloroform extracts of Xanthium strumarium.L roots treatment on marker
enzymes of EAC bearing mice
|
Groups (n)
|
ALT (IU/l)
|
AST (IU/l)
|
ALP (IU/l)
|
Total Bilirubin
|
|
Normal control
EAC control (1x106cells)
EAC + Xanthone
EAC + Xanthosine
EAC + Xanthine
EAC +
5FU(20mg/kgbw)
5- Fluorouracil
standard drug
|
181.5±5.06
292.00±8.75*
214.00±4.93**
224.50±5.19**
214.00±7.16**
208.75±8.26*
|
63.42±3.44
202.62±9.30*
102.00±3.12**
104.13±7.19**
105.54±5.14**
98.75±4.27*
|
221.50±5.45
271.50±6.30*
186.25±9.22**
211.25±11.21**
202.25±5.26**
190.00±11.37*
|
0.56±0.02
4.88±0.10*
1.32±0.17**
1.22±0.16**
1.73±0.13**
1.05±0.06*
|
|
ANOVA
f
df
|
39.3
5,18
0.0001
|
72.9
5,18
0.0001
|
12.3
5,18
0.0001
|
512.4
5,18
0.0001
|
*p values are expressed as mean ± SEM when n=6 rats in
each group.
*P< 0.001 compared to normal; **p < 0.001
compared to tumor control
Fig-7: Effect of Extracts of Xanthium strumarium L.
roots treatment on marker enzymes of EAC bearing mice.
ALP (IU/l)
Total Bilirubin
ALT (IU/l)
AST (IU/l)
Fig-8: Effect of Isolated Constituents from chloroform
extracts of Xanthium strumarium. L roots treatment on marker enzymes of
EAC bearing mice
ALT (IU/l)
AST (IU/l)
ALP (IU/l)
Total Bilirubin
The best parameters for gauging the effectiveness of
any anticancer agents are the prolongation of life span of experimental animals
13, 14. The reduction of
tumor volume viable total cell count finally reduces the burden and amplified the
life span of EAC bearing mice.
A regular rapid increase in ascetic fluid tumor volume
was observed in EAC tumor bearing mice. The tumor cells directly draw the
nutrition from ascetic fluid; it does mean the ascetic fluid continuously
supply the nutritional requirement to tumor cells15. Treatment with
chloroform extract suppressed the tumor volume, viable cell count and enhanced
the life span of the tumor bearing mice. It can be concluded that the
chloroform extract and isolated compounds of chloroform extract reducing the
nutritional fluid volume and inhibiting the tumor growth increases the life
span of EAC-bearing mice.
The major problems in the cancer chemotherapy are
myelosupression and anemia 16. Mainly the reduction of RBC and Hb% in
tumor bearing mice lead to the formation anemic condition and this may occur
either due iron deficiency or due to
hemolytic or myelopathic conditions17. The treatment with X. strumarium lignans restored the
hemoglobin (Hb) count, RBC and WBC count more or less to normal levels. This
finding clearly indicates that root extracts of X. strumarium posses effective protective properties on the
hemopoietic system.
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