PROTECTIVE EFFECT OF CURCUMIN ON HEMATOLOGICAL
ALTERATIONS INDUCED BY REPEATED ADMINISTRATION OF
FLUORIDE AND λ-CYHALOTHRIN ALONE AND IN COMBINATION
IN WISTAR RATS
Imtiyaz Ahmad,Rajinder Raina*,Pawan Kumar Verma and Sandeep Pandita
Division of Veterinary Pharmacology and Toxicology, Faculty of Veterinary Sciences and
Animal Husbandry, SKUAST-J, R.S. Pura, Jammu – 181102, India.
ABSTRACT
Present study was aimed to investigate protective effect of curcumin on
oral exposure of λ-cyhalothrin (LCT) and fluoride (F) on
haematological parameters in wistar rats. Forty eight rats were divided
into 8 groups with 6 rats in each. Animals in the Group I and II served
as control and received only normal tap water and curcumin
(100mg/kg, BW) respectively. The animals of Group III and IV were
provided drinking water containing F (10 ppm) and LCT at the dose
rate of 10 mg/kg BW through oral gavage respectively. Group V
received both the toxicants simultaneously. The animals of group VI
were treated with curcumin and provided water containing F and group
VII received curcumin along with LCT. Group VIII received curcumin
and both F in their drinking water and LCT daily for 14 days. Oral
administration of F or LCT alone for 14 days didn’t significantly alter
the haemoglobin Hb, packed cell volume (PCV), total erythrocyte count (TEC) levels but
co-exposure of toxicants for 14 days significantly (P<0.05) alters hematological parameters.
Co-exposure of toxicants also significantly (P<0.05) induced leucocytosis as compared to control
animals. Treatments with curcumin protect the hematological alterations induced by repeated
oral exposure of F and LCT for 14 days.
KEYWORDS: λ-cyhalothrin; haematological parameters; fluoride, curcumin.
Volume 4, Issue 3, 1231-1238. Research Article ISSN 2277– 7105
Article Received on 24 Dec 2014,
Revised on 18 Jan 2015, Accepted on 12 Feb 2015
*Correspondence for
Author
Prof. Rajinder Raina
Division of Veterinary
Pharmacology and
Toxicology Faculty of
Veterinary Sciences and
Animal Husbandry R.S.
Pura, Jammu - 181 102,
INTRODUCTION
λ-cyhalothrin [α-cyano-3- phenoxybenzyl - 3- (chloro-3, 3, 3-trifluoro - 1- propenyl)-2,
2-dimethylcyclopropanecarboxylate] a new synthetic type II pyrethroid used worldwide in
agriculture, pest control, protection of foodstuff and vector control.[1] Pyrethroids are reported
to generate free radicals through hydrolytic ester cleavage and oxidative pathways by the
CYP-450 enzymes.[2-5] Alteration of haematological parameters has been reported with
pyrethroids such as deltamethrin, bifenthrin, fenvalerate, chlorpyrifos and λ-cyhalothrin
(LCT).[6-10] Exposure to toxic metals and metalloids remains a widespread problem. High
accumulation of fluoride (F) in bones affects the formation of haemopoietic cells in bone
marrow cavities. It has been reported that F toxicity has adverse effects on blood and blood
constituents in different animal species. It causes anaemia due to decreased levels of Vitamin
B12, folic acid and iron. The decrease in Hb, total erythrocyte count (TEC), packed cell
volume (PCV) and mean corpuscular volume (MCV) has been reported in experimental
animals.[7-11]
Turmeric (Curcuma longa L.) is a medicinal plant extensively used as home remedy for
prevention and treatment of various diseases[12] and it is commonly used as a food additive
(spice) preservative and coloring agent in Asian countries. The yellow colour of turmeric
powder is due to curcumin which is chemically diferuloylmethane. Curcumin is also reported
to have antibacterial,[13] anti-tumor,[14] antioxidant[15] and anti-carcinogenic effects.[16-18]
Curcumin also scavenges the free radicals or inhibit the generation of reactive oxygen species
likesuperoxide anions, H2O2 and nitrite radical.[17] Due to presence of tremendous antioxidant
potential[15] its application in preventing diseases associated with oxidative damage has been
established in experimental animals and human trials.[19-20]
Increasing use of LCT and ever increasing F levels in drinking water, simultaneous exposure
of human and animals to these chemicals is a reality. Studies have shown that LCT increases
the F toxicity on simultaneous exposure in experimental animals primarily due to reduced
antioxidant defense or increased production of reactive oxygen species (ROS) due to
metabolism of xenobiotics [21-24]. Therefore present study was aimed to investigate protective
effect of curcumin on haematological alterations induced by repeated exposure of
MATERIALS AND METHODS
The effects of LCT and F alone and in combination were observed on apparently healthy
wistar rats of either sex weighing 150-200 g procured from Indian Institutes of Integrative
Medicine (Council of Scientific & Industrial Research, Lab), Jammu. The animals were
provided standard pelleted ration and clean drinking water ad libitum. All the animals were
maintained under standard managemental conditions. A daily cycle of 12 h of light and 12 h
of darkness was provided to animals. Prior to start of experiment, the rats were acclimatized
in the laboratory conditions for a period of more than 3 weeks. All the experimental animals
were kept under constant observation during entire period of study. The experimental design
was approved by the Institutional Animal Ethical Committee.
Experimental design: After acclimatization forty eight wistar rats were randomly divided into
eight groups with six rats in each. Animals in the Group I and II served as control and
received only normal tap water and curcumin (100mg/kg, BW, orally) respectively. The
animals of Group III and IV were provided drinking water containing F (10 ppm) and LCT at
the dose rate of 10 mg/kg bw through oral gavage respectively. Group V received both the
toxicants simultaneously i.e. F (10 ppm) and LCT (10 mg/kg, BW). The animals of group VI
were treated with curcumin and provided water containing F (10ppm) and group VII received
curcumin along with LCT (10mg/kg) through oral gavage. Group VIII received curcumin and
both F (10ppm) in their drinking water and LCT (10mg/kg) daily for 14 days. In order to
minimize the possible instability, both toxicants were prepared freshly in water. All the rats
were weighed weekly to make necessary corrections in the LCT dosage as per body weight.
Sample collection and analysis: After 14 days of daily treatment with curcumin, F and LCT
alone and in combination group blood samples were collected from retro orbital plexus using
capillary tubes in aliquots containing heparin as anticoagulant. TEC, TLC and PCV were
estimated by using the standard procedure of Rodak[25] and Hb was estimated by the methods
described Van kampen and Zilista.[26] The results are presented as mean ± standard error of
mean and to measure the level of significance, paired‘t’ test was applied.[27]
RESULTS
The effects of curcumin on hematological alterations (hemoglobin, PCV, TLC and TEC)
induced by repeated administration of LCT and F alone and in combination in wistar rats
were presented in table. Administration of either F or LCT for 14 days did not significantly
(P<0.05) reduced the hemoglobin concentration as compared to control and curcumin alone
exposed group. Repeated treatment with curcumin along with exposure of both the toxicants
restored the level of hemoglobin i.e. significantly (P<0.05) high from the toxicants exposed
group. Repeated treatment with F and LCT alone and in combination significant (P<0.05)
decreases the PCV (%) in rats. However, co-administration of both the toxicants induced
more significant decrease as compared to control group. Simultaneous exposure of curcumin
and toxicants in alone and in combination group significantly (P<0.05) restored the level of
PCV and TLC. A significant (P<0.05) reduction in mean values of total erythrocyte count
(x106/mm3) in all groups of animals were observed. The reduction in TEC was more
pronounced in co-toxicants exposed group as compared to exposure of either toxicant.
Treatment with curcumin fails to restored the reduced levels of TEC and remain significantly
(P<0.05) lover from the control group.
Table: Effect of repeated oral administration of curcumin on hematological alterations
(Hb, PCV, TEC and TLC) induced by repeated administration of F (fluoride) and LCT
(λ-cyhalothrin) alone and in combination in wistar rats.
Groups Hemoglobin
(gm/dl) PCV (%)
TEC (×106/mm3)
TLC (×103/mm3) Control 11.32b ± 0.22 37.98b ± 0.63 8.49b ± 0.20 9.39a ± 0.04 Curcumin 12.93b ± 0.44 41.11b ± 0.80 9.01c ± 0.10 9.19a ± 0.06 F (10 ppm) 10.87b ± 0.36 37.74b ± 0.32 7.92b ± 0.11 12.70c ± 0.24 LCT (10mg/kg) 10.84b ± 0.25 36.61b ± 0.79 7.46a ± 0.19 13.22c ± 0.17 F (10ppm) + LCT (10mg/kg) 9.76a ± 0.27 34.99a ± 0.66 7.15a ± 0.08 11.57b ± 0.34 Curcumin + F (10 ppm) 11.00b ± 0.41 39.69c ± 0.35 8.21b ±0.06 9.62a ± 0.17 Curcumin + LCT (10mg/kg) 10.96b ± 0.33 37.19b ± 0.60 8.19b ± 0.05 9.89a ± 0.37 Curcumin + F (10 ppm)
+ LCT (10mg/kg) 10.80
b
± 0.27 36.28b ± 0.19 7.92b ± 0.07 11.01b ± 0.14
Values given are mean ± SE of the results obtained from 6 animals.
Means with common superscript in a column do not differ significantly at 5% (P<0.05) level of significance.
Treatment with either toxicant alone significantly (P<0.05) increased the level of TLC in
animals as compared to control animals. Treatment with curcumin following repeated
exposure of either toxicant significantly restored the levels of TLC but co-exposure of both
the toxicant along with curcumin failed to restored the levels of TLC.
DISCUSSION
Production of excessive free radicals or reactive oxygen species (ROS) is responsible for the
capacity by scavenging free radicals or ROS at neutral and acidic pH.[20, 28-29] Indiscriminate
use of pesticides and increasing levels of metal and metalloids in soil and water are main
source of contamination to human and animals. Ingestion or inhalation of these contaminants
might be responsible for subtle health effects due to disturbance in oxidant and antioxidant
status of the mammals. Curcumin alone treatment didn’t induced any significant difference in
the values of various haematological parameters as compared to control values following 14
days of exposure. Significant reduction in Hb and TEC on co-exposed group may be due to
formation of Heinz bodies during pesticide exposure resulting from the conversion of
hemoglobin to methemoglobin. The attachment of Heinz bodies to the plasma membrane
increases membrane rigidity leading to increased red blood cell (RBC) lysis or its premature
removal from circulation.[30] Additionally the significant reduction in the TEC may be
contributed by the effect of these (F or LCT) toxicants in haemopoietic tissue. This is
especially in the light of studies that have shown the ability of pesticides to be toxic to the
immune cells via the induction of necrosis and apoptosis.[31] Similar observations were also
reported by following repeated λ-cyhalothrin exposure in rats.[32]
Reduction of Hb, TEC, PCV and increased TLC levels in co-exposed group indicative of
anaemia observed in rats exposed to toxicants may be due to ability of these to decrease
tissue iron concentration,[30] interference with Hb biosynthesis and shortening of RBC
lifespan or even increase in erythrocyte fragility. Further significant reduction in PCV on
exposure of F and LCT in present study indicates sign of anemia.[30] These changes may be
due to decrease in activities of bone marrow and myelosuppression.[33]
Similarly, treatment with curcumin along with toxicants (F and LCT) showed no significant
difference on Hb concentration as compared to control following 14 days of exposure in rats.
The pretreatment with curcumin may protect Hb from oxidation which was depressed by
toxicants.[34] Glutathione is one of the most free radical fighters in the human body and
curcumin increases the expression of intracellular glutathione. Glutathione interact with ROS
or free radicals and get converted into oxidized form thus protect from damaging effect.[35]
Furthermore the ability of curcumin to improve the absorption of iron from the gut or by
facilitating the reduction of oxidized iron to its reduced form.[8-10,36]
CONCLUSION
Oral administration of F or LCT alone for 14 days didn’t significantly alter the Hb, PCV,
parameters. Treatments with curcumin protect the hematological alterations induced by
repeated exposure of F and LCT for 14 days.
ACKNOWLEDGMENTS
Authors thanks to the Dean, Faculty of Veterinary Sciences and Animal Husbandry,
SKUAST-Jammu for providing necessary facilities for conducting the research.
Conflict of interest: Nil
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