Iraqi Journal of Veterinary Sciences, Vol. 37, No.

4, 2023 (775-780)

Iraqi Journal of Veterinary Sciences

Ameliorative effect of nano chromium on some hematological values and

GLUT-2 in rabbits fed a high- Fat diet
H.M. Hameed
Department of Physiology, Biochemistry and Pharmacology, College of Veterinary Medicine, University of Mosul, Iraq

Article information Abstract

Article history: Nano-chromium has an anabolic effect when used in low concentrations and boosts
Received 25 October, 2022
Accepted 05 May, 2023 nucleic acid synthesis in the liver; it is also termed a glucose tolerance factor (GTF)
Available online 23 August, 2023 because it regulates the ' 'insulin's metabolic action. It is used in the metabolism of
carbohydrates,
proteins, and fats. This study was carried out on 20 local white male rabbits (aged two
Keywords: months), which were randomly assigned into four groups: the first was a control group fed
Nano-chromium
Rabbits
on a standard diet, the second was a group fed on a high-fatty diet (HFD), the third was a
GLUT-2 group fed on a standard diet which has was given Chromium at a concentration of 2 mg/L
Insulin with drinking water, and the fourth was a group of nano chromium fed on a fatty diet who
Glucose were given chromium 2 mg/ L of drinking water. The fatty diet group had higher levels of
Correspondence:
erythrocytes, leukocytes, Hb, hematocrit, lymphocytes, neutrophil, glucose, cholesterol,
H.M. Hameed triglyceride, GLUT-2, insulin, and insulin resistance with a decrease in HDL. In
hadeel.mohammad@uomosul.edu.iq comparison, the two groups of chromium nanoparticles showed lower levels of erythrocytes,
leukocytes, Hb, hematocrit lymphocytes, neutrophil, glucose, cholesterol, triglyceride
GLUT-2, insulin, and insulin resistance but higher levels of HDL. Our objective is to
investigate the deleterious effects of the HFD on hematological values, GLUT-2 levels,
and insulin levels, as well as the potential for nano-chromium to enhance these qualities.
DOI: 10.33899/ijvs.2022.136659.2604, ©Authors, 2023, College of Veterinary Medicine, University of Mosul.
This is an open access article under the CC BY 4.0 license (http://creativecommons.org/licenses/by/4.0/).

Introduction greater material sizes, enhancing biocompatibility thus,

nanotechnology can be used to feed animals to provide better
Over several years, many materials have been used as nutrients improving the immune status of animals and raising
feed additives in animal diets, including enzymesprobiotics, the efficiency of animal production (7-9). Nanotechnology
and organic acid (1,2). However, as a result of the rapid also provides a wide field for veterinarians for treatment,
development in the field of animal production in recent diagnosis, tissue engineering, and the production of modern
years, nanoscale has gotten great attention within the vaccines and disinfectants, as it can already be used in
scientific communities in many countries, where many of animal care, production, breeding, and reproduction (10).
the companies specializing in the manufacture of new Nanocomposites are manufactured in different forms,
shapes of nanoscale resources that concern animal systems including the production of metallic nanoparticles, as their
of manufacturing to boost production efficiency (3,4). The production provides many advantages for the production of
word nano is derived from the Latin phrase Nanos, which animal feed at a lower cost and lower concentrations to be
implies midget the sizes of nanoparticles range from 1- used as potential alternatives to feed additives instead of
100nm (5,6). As a result of the special properties of antibiotics, they can also help in controlling the microbes
nanoparticles represented by their small size, which allows sitting in the meal and regulating the digestion and
them to pass easily through the walls of the alimentary absorption process (11). One of these minerals is nano-
canal and into the cells of the body faster than usual chromium, using nanocomposites as feed additives instead
molecules with
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 Iraqi Journal of Veterinary Sciences, Vol. 37, No. 4, 2023 (775-780)

of large molecules reduces the excretion of minerals (12). heparinized tubes to conduct blood analyses, and the other
Chromium is a compound with a small molecular weight
and low toxicity that can increase insulin activity and slow
the progression of diabetes (13,14). Cr is a crucial trace
mineral required for most vital metabolic processes. It is
used in the metabolism of carbohydrates, protein, and fats
and has anabolic effects during animal exposure to various
types of stress (15,16). Chromium is also an important
micronutrient required for glucose metabolism, and its
deficiency causes high blood sugar and insulin resistance
(17-18). Chromium supplementation has been confirmed to
enhance carbohydrate metabolism by potentiating and
lowering insulin resistance signifiers, lipid variables, and
blood insulin levels (19,20).
Our objective is to investigate the deleterious effects of
the HFD on hematological values, GLUT-2 levels, and
insulin levels, as well as the potential for nano-chromium to
enhance these qualities.

Materials and

methods Ethical

approve
The University of Mosul, College of Veterinary
Medicine's Animal Care and Use Committee evaluated and
approved each step of the experiment's protocols.

Rabbits and diets

In most developing countries, where animal protein
supply is insufficient to fulfill the population's protein
needs, rabbit offers much potential to assist meet these
needs; this is because it has a clear edge over other animals,
rabbits are known to reproduce swiftly and have the
capacity to eat a variety of foods (21). This treatise was
conducted in the experimenter animal house of the College
of the Veterinary Medicine / University of Mosul; 20 male
rabbits were used at the age of 2 months. The current study
lasted for two months, during which the animals were fed
standard and provided free feed and water. The nano-Cr
powder was provided by 3302Twing Leaflane, Houston,
TX77084, USA.

Experimental design
In this reserch, 20 male rabbits were used, which were
divided random into four groups (5 rabbits/group) as
follows; The control fed a standard diet (control), the group
fed a high fatty diet (HFD), a group of nano-chromium fed
on a standard ration with nano-Cr two mg/L with drinking
water and a group of nano-chromium fed on a high fatty
meal with nano-Cr 2 mg/ml with drinking water
(HFD+Nano-Cr) (22).

Blood sample collection

Blood specimens were obtained across the marginal ear
vein after the animals have fasted overnight, and the
samples were split into two groups; the first was situated in
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 Iraqi Journal of Veterinary Sciences, Vol. 37, No. 4, 2023 (775-780)

section has been put in tubes without anticoagulants

which were left to clot at room temperature and
discarded in a centrifuge with a capacity of 3000 rpm to
obtain serum, the samples were kept at -20°C until
laboratory analyzes were carried out.

Studied standards
Blood samples containing anticoagulants were used
to conduct hematological tests represented in the number
of erythrocytes (RBC) and leukocyte(WBC), differential
leukocyte count(DLC) hemoglobin (Hb),Hematocrit%,
MCV, MCH and MCHC (23), while serum samples were
used to measure glucose level using the ready-made
analysis kit manufactured by (Randox Company) and the
estimation of the level of cholesterol, triglycerides, high-
density lipoproteins – cholesterol (HDL-C), using the
ready-made kit manufactured by a French company
(Biolabs) and calculating the atherogenic index in
addition to estimating the level of glucose-2
transporter(GLUT-2) and insulin level in the blood
serum using ELISA (24).

Statistical analysis
Statistical analysis of experimental data was carried
out using a one-way analysis of variation, and
distinctions among groups were determined using
Duncan's test for all criteria covered in the study (25).

Results

It is noticed from the data obtained that there was a

stastical rise (P≤0.05) in the No of erythrocytes,
leukocytes, hemoglobin, and hematocrit for the group of
fat diet compared with the control. The treatment with
nano-Cr led to a decline in the blood parameters
compared with the control, while the group of nano-Cr
with the fatty diet showed a decrease in the above
parameters compared with the fatty diet. The treatment
with nano-Cr improved the negative effect of the fatty
diet by restoring the hematological parameters to their
normal level, which is close to the control (Table 1).
Giving the rabbit a fatty diet led to a significant rise
in lymphocytes, neutrophils, and basophils, with a
reduction in eosinophils and monocyte compared to the
control. The chromium nanoscale group showed a
decrease in lymphocytes and monocyte accompanied by
an enhance decrease in lymphocytes and an enhance
decrease in lymphocytes and an enhancement decrease in
lymphocytes and monocyte accompanied by an
enhancement in eosinophils in comparison to the control
group, The addition of chromium nanoform to the fatty
diet, led to a decline in lymphocytes and neutrophils with
a rise in eosinophils and monocyte in comparison to the
fatty diet group. The use of nano-Cr improved the
unwanted effect of the fatty diet (Table 2).

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 Iraqi Journal of Veterinary Sciences, Vol. 37, No. 4, 2023 (775-780)

The study indicates a significant reduction in MCV and nanoform improved the negative aspects of the fatty diet by
MCH with a rise in MCHC for the fatty diet group returning the parameters to their normal level, which was
compared with the control. Chromium nanoparticles close to the control group (Table 4).
showed a rise in MCV and a decrease in MCHC in The data indicated a significant rise in LDL, VLDL, and
comparison to the control. The addition of chromium atherogenic index, accompanied by a reduction in HDL for
nanoparticles to the fatty diet led to an increase in MCV the fatty diet group in comparison with the control group.
and MCH with a decrease in MCHC compared to the fatty The Chromium nanosized showed a decrease in LDL,
diet group (Table 3). VLDL, and atherogenic index with a rise in HDL compared
The results indicate a significant elevate in the glucose, to the control. Compared to the fatty diet group, adding nano-
cholesterol, and triglycerides of the fatty diet group in chromium to the fatty diet resulted in a decrease in LDL,
comparsion with the control. The addition of chromium VLDL, and the atherogenic index, as well as an increase in
nanoscale to the ratio led to a decline in the biochemical HDL, indicating the importance of nano-chromium in
parameters compared to the control. The group of nano-Cr improving the negative impact of the fatty diet (Table 5).
with the fatty diet showed a significant reduction in the
above parameters compared to the fatty diet. The chromium

Table 1: Effect of nano-Cr on some hematological value in rabbits given a high-fat diet

Treatment RBC (106 /mm3) WBC (103/mm3) Hematocrit (%) Hb (g/dl)

Control 5.86±1.96 b 6.48±2.71 b 30.60±3.40 b 10.95±2.01 b
HFD 6.93±2.81 a 7.46±3.89 a 31.98±4.29 a 12.38±3.51 a
Nano-Cr 4.99±1.66 c 5.01±2.87 c 29.73±3.01 c 9.19±3.22 c
HDF+ Nano-Cr 4.01±1.52 d 4.93±2.75 d 27.41±2.87 d 8.26±3.17 d
The information is presented as an average± SE. Little distinct letters in the column indicate the major difference between
collections at P≤0.05.

Table 2: Effect of nano-Cr on DLC in rabbits given a high-fat diet

Treatment Lymphocyte (%) Neutrophil (%) Eosinophil (%) Basophil (%) Monocyte (%)
Control 45.15±0.78 b 37.21±0.39 b 1.96±0.10 c 1.91±0.16 a 13.85±1.40 b
HFD 51.62±0.92 a 40.17±0.47 a 2.47±0.18 b 1.09±0.10 a 10.11±1.27c
Nano-Cr 43.88±0.71 c 36.98±0.34 b 3.68±0.20 a 1.89±0.14 a 13.93±1.44 b
HDF+ Nano-Cr 42.94±0.68 c 35.98±0.31 c 3.87±0.25 a 1.88±0.14 a 14.23±1.52 a
The information is presented as an average± SE. Little distinct letters in the column indicate the major difference between
collections at P≤0.05.

Table 3: Effect of Nano-Cr on blood indices in rabbits given a high-fat diet

Treatment MCV (fl) MCH (pg) MCHC (g/100ml)

Control 52.25±2.11 c 18.77±2.33 b 35.51±2.12 b
HFD 47.23±2.09 d 17.82±1.32 c 38.58±2.78 a
Nano-Cr 59.62 ±2.29 b 18.89±2.45 b 30.94±1.31 c
HDF+ Nano-Cr 68.39±2.98 a 21.09±2.88 a 30.29±1.18 c
The information is presented as an average± SE. Little distinct letters in the column indicate the major difference between
collections at P≤0.05.

Table 4: Effect of nano-Cr in glucose and cholesterol and triglyceride in rabbits given a high-fat diet

Treatment Glucose (mg/100ml) Cholesterol (mg/100ml) Triglyceride (mg/100ml)

Control 116.35±2.98 c 332.61±3.57 b 99.67±1.82 b
HFD 387.61±3.84 a 529.43±4.93 a 120.89±2.76 a
Nano-Cr 2mg/L 114.19±2.75 d 319.75±3.11c 88.54±2.09 d
HDF+ Nano-Cr 2mg/L 121.87±2.48 b 310.82±2.39 d 94.49±1.65 c
The information is presented as an average± SE. Little distinct letters in the column indicate the major difference between
collections at P≤0.05.

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Table 5: Effect of nano-Cr on lipoprotein value in rabbits given a high-fat diet

Treatment HDL-C (mg/100ml) LDL-C (mg/100ml) VLDL-C (mg/100ml) Atherogenic index

Control 28.93±1.69 c 263.48±2.70 b 19.35±2.11 b 11.78±1.85 b
HFD 19.76±1.09 d 495.49±3.05 a 26.18±2.79 a 26.66±1.98 a
Nano-Cr 30.64±2.19 b 213.41±2.08 d 18.71±2.04 c 10.48±1.67 c
HDF+ Nano-Cr 33.59±2.97 a 248.53±2.14 c 17.98±1.73 d 9.28±1.18 d
The information is presented as an average± SE. Little distinct letters in the column indicate the major difference between
collections at P≤0.05.

Compared to the control, the fatty diet group had a corresponds to what was stated by Dworzański et al. (29),
significant rise (P0.05) in Glut-2, insulin, and insulin Amer et al. (30) if they implied the nano chromium reduction
resistance. Compared to the control, nano chromium in leukocyte in the case of using a fatty diet. The addendum
treatment reduced Glut-2, insulin, and insulin resistance. of Cr to the ratio of rabbits at a concentration of 0.6 mg/kg
The addition of nano-chromium to the fatty diet also of food led to a decrease in the value of cholesterol, LDL,
reduced the above parameters compared to the fatty diet TG, and an increase in HDL. Nano- Chromium plays a
group, demonstrating the role of nano-chromium in crucial role in preserving blood glucose (31). The improved
improving the fatty diet's negative effect (Figure 1). effect of Chromium can be attributed to its role as an
antioxidant mineral, which increases the level of
glutathione, superoxide dismutase, and catalase inside the
body (32). Numerous studies have shown that Chromium
can act as an immunomodulator (stimulating or
suppressing) by influencing immune system elements such
as B cells, T cells, and macrophages, as well as cytokine
production (33). A lack of this mineral in the body results in
weakened humoral immunity (34). Trace elements, like
metals, are necessary for the proper operation of
biochemical processes, particularly as cofactors for
enzymes, and in a wide range of methods implicated in
glucose assimilation and hormonal control, particularly
insulin, Chromium is a mineral that the body necessitates in
Figure 1: Effect of nano-Cr on GLUT-2 and insulin and trace quantities and is among the most broadly used and
insulin resistance in rabbits given a high fat. overall nutrition (35). Chromium is an insulin cofactor that
enhances amino acid absorption, promotes lipogenesis from
Discussion glucose, and promotes fat storage in the liver and fatty
tissues (36,37). This allows Cr to participate in the
The current study investigated the hypothesis of using a autoamplificatione of insulin signaling to maintain the
fatty diet. Its effect on some blood traits, insulin, and active conformation, which improves glucose tolerance by
GLUT- 2 levels, and the possibility of improving them by potentiating the action of insulin, which is required for
using nano-chromium as feed additives to the rabbit diet, nutrient metabolism (38). Accordingly, to a previously
was put forward, as fatty food can cause wide inflammatory described hypothesized mechanism of action for Chromium
effects resulting in a boost in the rate of white cells and in insulin control, Chromium improves insulin governing to
neutrophils (26), and this is consistent with Maysami et al. target tissue by raising the sensitivity and quantity of
(27) where he stated that an excessive amount of fat in the insulin receptors, which are found in most cells but vary in
meal could cause not only an exponential rise in the rate of concentration depending on cell type. Insulin receptors are
leukocyte but also to a significant rise in erythrocyte, MCV, made up of two extracellular α subunits that include the
hemoglobin and PCV value. The fatty tissue system is insulin-bounding site and two trans membrane β subunits,
linked to a variety of health issues, including the occurrence thus Chromium, like insulin, appears to impact protein
of insulin resistance; that obesity is characterized by an phosphorylation-dephosphorylation events, according to
increased fatty acid backup in the extending fat tissue structural characteristics (39). Phosphotyrosine phosphatase
quantity and is tightly linked to the onset of insulin (PTP-1) disables the insulin receptor, and Cr inhibits it.
resistance in peripheral tissues such as adipose and skeletal Both of Chromium's effects would result in enhanced
muscle (28). The use of nano- chromium led to reversing phosphorylation of the insulin receptor, which is linked to
the negative effects of a fatty diet if the results showed a greater insulin sensitivity, resulting in much lower glucose
decrease in blood parameters, lipid profile, insulin level, levels and, as a result, an improvement in the
and insulin resistance, and this hyperglycemic state (40). Thus, the improved effect of

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nano-Cr can be

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