Egypt. J. Comp. Path &Clinic Path. Vol. 28 No.1, 2015 ; 103- 116 ISSN 1110-7537

103

Clinico-pathological Studies on The Regulatory Effect

of Cinnamon on Metabolic Disorders in High

Fructose-Fed Mice

Walaa M.S. Ahmeda, Nermeen A. Helmy

b and Taghreed M.Nabil

c

a. Department of Clinical pathology, faculty of veterinary medicine, Beni-suef university.

b. Department of Physiology, faculty of veterinary medicine, Beni-Suef university.

c. Department of Cytology and Histology, faculty of veterinary medicine, Beni-Suef university.

ABSTRACT— In the current study we investigate the possible effect of cinnamon against

some metabolic changes induced by high fructose intake in mice. For this purpose, eighty mice

were divided into four groups, the first one kept as control. In the second group, mice were given

high fructose (20%) in water per day for 8 weeks. In the third and fourth groups, mice were

given fructose as in the second group and orally administrated cinnamon daily for 8 weeks in a

dose rate of 150 and 300 mg/kg b.w, respectively. Body weights were recorded weekly and oral

glucose tolerance test was carried out at the end of experiment. Hepatic enzymes (ALT, AST and

ALP), glucose, lipid profile (total cholesterol, triglyceride, HDL-c and LDL-c), TNF-α levels

were estimated. Mice liver of were weighted and its antioxidant enzymes (GSH, CAT) content

and lipid peroxidation (MDA) level were measured. Results indicate no change in hepatic

enzymes in fructose-fed mice. Co-administration of cinnamon with fructose resulted in decrease

the elevated glucose and TNF-α values. Moreover, cinnamon improve the disturbed lipid profile.

The depletion of antioxidant enzymes and increased MDA activity by fructose intake were

ameliorated in cinnamon treated mice. Examination of liver by light and electron microscopy

revealed fatty change, cloudy swelling, inflammatory cells infiltration in fructose-fed mice which

were markedly improved by cinnamon treatment at dose of 150 mg/kg. In conclusion, co-

administration of cinnamon at dose of 150 mg/kg was more effective than the higher dose and

regulate the disturbed metabolic changes that induced by high fructose intake in mice.

Key words: Cinnamon; High fructose- diet; Histopathology; Metabolic disorders; Mice.

—————————— ——————————

INTRODUCTION

Nutritional overfeeding of fat and or

refined carbohydrates is a contributing factor

to the development of a complex pattern of

disorders that include insulin resistance,

dyslipidemia, diabetes, obesity,

atherosclerosis, and cardiovascular disease

(Marissal-Arvy et al. 2014). Glucose and

lipids are key components in energetic

metabolism. Excess energy intake is

responsible for an inappropriate processing of

the glucose and lipid. This extreme energy

will stored in adipocytes which produce high

levels of adipokines and inflammatory

cytokines that affect the vascular system

(Galic et al. 2010). Some of these cytokines

(tumor necrosis factor interleukin-1band and

interleukin-6) could be involved in initiating

of insulin resistance (Bastard et al. 2006).

To gain insights into the association between

diets and these diseases and develop effective

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therapy, animal models have been developed

that typically rely on genetic manipulation,

toxic injury [e.g., methionine- and choline-

deficient diet], or dietary extremes (e.g.,

abnormally high-fat or high-fructose diets)

(Anstee and Goldin, 2006).The metabolism

of fructose is done by the liver. High fructose

intake develop metabolic consequences such

as hyperinsulinemia, insulin resistance,

dyslipidemia, hyperuricemia, inflammation,

oxidative stress, and disrupted status of

adiponectin, leptin and tumor necrosis factor-

a (Kitagawa et al. 2012). Most of

conventional drug treatment of insulin

resistance have undesirable side effects such

as weight gain, fluid retention, and an

increased risk of myocardial infarctions.

Whilst, medicinal plants are expected to have

a similar degree of efficacy without the

troublesome side effects associated with

pharmacological treatments (Eddouks et al.

2015).

Dietary botanical supplements are a

valuable source of therapeutics for preventing

epidemic diseases such as obesity,

cardiovascular disease and diabetes (Cheng

et al. 2012). Cinnamon, one of the widely

used flavoring agent and medicinal plant

belonging to the family Lauraceae, possess a

significant anti-bacterial, anti-fungal, anti-

inflammatory, antioxidant and anti-diabetic

properties (El Hasnaoui et al. 2015).

In the present study, we have studied

the impact of high-fructose consumption and

elucidated the influence of cinnamon on the

metabolic alterations induced by high-

fructose intake in mice.

MATERIALS AND METHODS Fructose was obtained from (Sigma

company, USA), and was prepared in 20 %

solution.

Cinnamon bark (Cinnamomum zeylanicum)

was purchased from the local market.

Glucose, Alanine aminotransferase

enzyme (ALT), aspartate aminotransferase

enzyme (AST), alkaline phosphatase enzyme

(ALP) reagent kits were obtained from

Biodiagnositic Company, Egypt. Total

cholesterol (TC), triglyceride (TG) and high-

density lipoprotein (HDL) was performed

using reagent kits from Spin react (Spain).

reduced glutathione (GSH), Catalase (CAT)

and malondialdehyde (MDA) were

estimated using commercial kits (Bio-

diagnostic for Research Kits, Egypt). All

the chemicals were of analytical grade

and were procured from local commercial

companies.

2.1. Study design

A total of 80 BALB/c mice were

purchased from Research Institute of

Ophthalmology, Financial management

(Giza, Egypt). The animals were handled and

cared in accordance with the guidelines of

Beni-Suef University for animal use. After 1

week of acclimatization, mice were randomly

divided into 4 groups (n=20/group). One

group was kept as negative control; a second

group was given 20 % fructose in water; a

third group was given 20 % of fructose along

with cinnamon in a dose of 150 mg/kg b.w; a

fourth group was given 20% fructose along

with cinnamon in a dose of 300 mg/ kg b.w.

The Cinnamon bark was finely

powdered in a mechanical mixer, weighted

then dissolved in water and kept in a water

bath at 60°C for two hours and filtered.

Cinnamon was administered orally to mice

once a day for two months. Body weight of

mice was determined weekly till the end of

the experiment.

Oral Glucose tolerance test (OGTT) The oral glucose tolerance test was

performed in the last day of the experiment

according to Yadav et al. (2007).

2.2. Sample collection

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From all groups, blood samples were

collected from the orbital venous plexus,

allowed to clot for 20 minutes and then were

centrifuged at 3000 rpm for 10 minutes.

Serum was separated and stored at -20 ºc till

analysis. The mice were euthanized by

cervical dislocation and liver was dissected

and weighted in order to calculate the

organ/body weight ratios for each mice.

Then, part of liver tissue was used for

histopathological studies. Another part was

prepared for estimation of GSH, CAT and

MDA according to Ahmed et al. (2014).

2.3. Laboratory analyses.

2.3.1. Serum hepatic enzymes. The activities of ALT and AST

enzymes were estimated with colorimetric

method (Reitman and Franel, 1975).

ALP activity was measured according to

Belfield and Goldberg (1971).

2.3.2. Blood glucose estimation Glucose level was estimated

according to Trinder (1969).

2.3.3 Lipid profile was determined by

measuring the content of total cholesterol,

triglycerides, HDL-c and LDL-c

concentrations with a standard colorimetric

method using reagent kits as indicated by

the manufacturer’s instructions.

2.3.4 Tumor necrosis factor -α (TNF-α)

Tumor necrosis factor α was

determined by in vitro Enzyme Linked

Immunosorbent Assay [ELISA] kit, using

colorimetric reaction method as instructed in

the kit manual.

2.3.5 Estimation of GSH, CAT, and lipid

peroxidation in liver tissue GSH and CAT activities were

determined by spectrophotometric method

(Beutler et al., 1963 and Aebi, 1984

respectively). Lipid peroxidation level was

measured by the thiobarbituric acid reaction

(Ohkawa et al., 1979).

2.3.6 Histopathological examination For light microscopic examination,

liver specimens were fixed Bouin's fluid,

dehydrated, cleared and embedded in

paraffin. Thin serial sections (5 μm) were cut

and stained with H&E for general studies and

Masson’s trichrome stain for collagen

demonstration. All the aforementioned

fixatives and stains as outlined by Bancroft

and Gamble (2008). For Transmission

electron microscopic (TEM) examination, the

collected liver specimens were cut in very

small pieces (1mm3) and fixed in 4%

gluteraldehyde solution (Hayat, 1986). They

were then post-fixed in 1% osmium tetroxide,

dehydrated and embedded in Epoxy medium.

Ultrathin sections were cut, stained with

uranyl acetate and lead citrate (Reynolds,

1963) and then examined by using

transmission electron microscope JEOL

(JEM-1400 TEM 80 kv) in the electron

microscope unit of faculty of Agriculture,

Cairo University.

2.4 Statistical analysis

All data are expressed in mean ±

standard errors. A statistical analysis was

performed by analysis of variance (ANOVA)

followed by a Tukey’s tests using GraphPad

Prism (GraphPad, San Diego, CA). For all

the statistical tests, the level of significance

was fixed at P< 0.05.

RESULTS Oral glucose tolerance test

In the OGTT (Fig. 1), the mean

values of fasting glucose was significantly

elevated in all treated mice when compared

to control. A rise in blood glucose level was

observed in all mice groups after 30 min

from glucose administration. The Fructose

fed mice exhibited delayed glucose clearance

and the glucose value was elevated at all the

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time points as compared to control. While,

treatment with cinnamon at dose of 150

(mg/kg) significantly enhanced glucose

clearance as compared with fructose-alone

given group. Administration of cinnamon in a

dose of 300 (mg/kg) significantly lowered the

glucose level compared with fructose group;

however they failed to reach to the normal

level.

Body and liver weight:

The changes in body weight of the

mice during the 8weeks did not differ

significantly between groups (Table 1 ). No

significant changes in the mice liver weights

and the ratio of liver weight to body weight

were noticed after 8 weeks in the fructose fed

group.

Serum hepatic enzymes.

The activities of ALT, AST and ALP

enzymes showed no significant changes

among all groups (Table 2).

Blood glucose estimation

The levels of serum glucose was

significantly higher in fructose-fed mice as

compared to that of control. Co-

administration with cinnamon at dose rate of

150 mg/kg significantly reduced the levels of

glucose when compare to normal value

(Table 2).

Lipid profile

As shown in Table (2), there were

significant differences in the measured serum

lipid profile between the fructose-treated

mice and control group. Serum

concentrations of the total cholesterol,

triglyceride and LDL-C were decreased in

the cinnamon- treated groups than in the

fructose group. However, TC and LDL-C

values fails to return to normal in both doses

cinnamon treated mice. The level of HDL–C

was significantly decreased in fructose group,

but it found to be significantly higher in the

cinnamon treated groups compared to

fructose group.

Tumor necrosis factor -α

In fructose-fed mice significant

increase was observed in TNF-α value as

compared to control (Table 2). Treatment

with cinnamon at dose rate of 150 mg/kg

prevented that increase and brought it back

the level to normal level. But, in mice group

that treated with cinnamon at dose of 300

mg/kg the TNF-α value failed to return to

normal.

Hepatic antioxidants enzyme and lipid

peroxidation.

The activities of antioxidant enzymes

are shown in Table (2). The mice in fructose

group exhibited a significant decrease in

hepatic GSH and CAT activities compared

with the control group. Co-treatment with

150 mg/kg of cinnamon caused a significant

increase of GSH and CAT activities

compared with the activity in mice fed the

fructose only. However, GSH activity in

cinnamon group at dose rate of 300 mg/kg

was non- significantly increased than fructose

group and still significantly lower as

compared to control.

Figure (1): Glucose tolerance test at different

time. The data are given as means ± standard

error (SE). C= control, fructose, F= Fructose;

FC 150= fructose +Cinnamon dose of 150

mg/kg and FC 300= fructose +Cinnamon dose

of 300 mg/kg.

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Table (1): Results of body weight from beginning of experiment till 8 weeks; liver weight

and ratio of liver weight to body weight of all mice groups.

The data are given as means ± standard error (SE), C= control, fructose, F= Fructose; FC 150= fructose

+Cinnamon dose of 150 mg/kg and FC 300= fructose +Cinnamon dose of 300 mg/kg.

Table (2): Effect of high fructose diet and cinnamon on glucose, hepatic enzymes, lipid

profile, TNF-α and hepatic antioxidant enzymes (M±SE).

The data are given as means ± standard error (SE), with dissimilar superscript letters (significantly differ at

p < 0.05): (a) letter is significantly differing from control value; (b) letter is significantly differing from

fructose group. n = 20/group. C, F, , FC 150 and FC 300 represent control, fructose, fructose +Cinnamon dose

of 150 mg/kg and fructose +Cinnamon dose of 300 mg/kg, respectively.

C F FC 150 FC 300

Body weight

(g)

1st week 20.00± 0.71 24.20±1.98 23.80 ±1.46 21.20±0.73

2nd

week 22.40±1.75 27.20±1.02 26.40±1.44 23.20±0.58

3rd

week 30.40±1.44 33.40±1.40 31.60±0.93 29.60±0.51

4th

week 31.60±1.54 34.00±1.52 31.20±0.86 32.40±0.75

5th

week 32.60±1.22 34.40±1.01 31.80±1.17 32.60±0.51

6th

week 33.00±0.92 35.20±0.98 32.40±0.93 33.20±0.87

7th

week 33.60±1.03 36.40±0.68 34.00±0.71 34.40±0.51

8th

week 34.50±0.68 37.30±0.85 35.40±0.40 34.60±0.68

Liver weight

Absolute (g) 1.58± 0.19 1.84±0.05 1.66±0.04 1.62±0.04

Relative (%)

4.58±0.50 4.93±0.16 4.69±0.11 4.68±0.05

Parameters C F FC 150 FC 300

Glucose (mg/dl) 103.7 ±5.6 135.9± 4.8a 106.6± 5.8

b 115.2± 4.6

AST (U/ml) 44.50±3.4 53.27±4.1 45.77±3.3 52.83±3.2

ALT (U/ml) 29.17±0.89 30.20±0.78 29.50±0.74 30.67±0.92

ALP (IU/l) 108.4±5.3 122.9±3.8 112.0±5.1 115.6±4.9

TC (mg/dl) 86.11±4.3 134.1±4.6 a 114.9±4.7

ab 116.8±4.1

a

TG (mg/dl) 72.65± 1.68 100.8± 3.59a 79.16± 3.27

b 82.65± 2.64

b

HDL-c (mg/dl) 49.3±3.5 35.8±2.3a 48.09±3.9

b 41.40±1.9

LDL-c (mg/dl) 22.31±4.87 78.04±3.38a 51.02±2.77

ab 59.02±4.24

ab

TNF-α (pg/ml) 27.44±0.80 43.03±1.97a 31.67±0.87

b 35.30±1.83

ab

GSH (U/g tissue) 2.87±0.04 1.27±0.22a 2.13±0.13

ab 1.76±0.09

a

CAT (U/g tissue) 1.82±0.15 0.88±0.08a 1.68±0.16

b 1.28±0.12

MDA (U/g tissue) 12.46±0.39 19.81±0.45a 11.19±0.33

b 11.83±0.75

b

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Histopathological examination

By light microscope, liver of control

showed normal architecture, the hepatocytes

(large, polygonal cells and some were

binucleated) arranged as hepatic cords

radiating from central veins and alternating

with blood sinusoids (Fig. 2.A). Stained

sections with Masson trichrome technique

revealed minimal amount of collagen fibers

around the central veins, hepatocytes and

periportal hepatic area (Fig. 2.B). With

electron microscope, the hepatocytes

cytoplasm contained rounded mitochondria,

well developed Golgi apparatus, rough

endoplasmic reticulum and smooth

endoplasmic reticulum. Their nuclei were

euchromatic with prominent nucleoli (Fig.

2.C). Marked histological changes were

observed in fructose group. The hepatocytes

showed degeneration (Fig. 3.A), marked fatty

change which exhibited by large vacuoles

pushing the nucleus toward the periphery of

the cell (Fig. 3.B) and cloudy swelling.

Degenerated hepatocytes with pyknotic

nuclei were seen and some cells had lost their

nuclei. Congested central vein; blood

sinusoids with von kupffer cells hyperplasia;

and diffuse and focal inflammatory cells

infiltration were recognized (Fig. 3.C). An

increase in the collagen fibers around the

central vein and in the periportal hepatic area

(Fig. 3.D).

With electron microscopy, the

hepatocyte revealed decrease in cytoplasmic

organelles, increasing fat globules and the

nuclei appeared heterochromatic (Fig. 3.E).

The liver of cinnamon at dose of 150 mg/kg

showed great improvement. However,

hyperplasia of von kupffer cells and

congested central vein were still noticed (Fig.

4.A). Collagen content became few (Fig.

4.B). Moreover, marked improvement was

seen in the hepatocytes by electron

microscopy. Reappearance of cytoplasmic

organelles and the nuclei were euchromatic

(Fig. 4.C). The liver of mice treated with 300

mg/kg of cinnamon were incompletely

improved. Most of hepatocytes surrounding

the central vein appeared with normal

histological profile. While that located more

peripherally showed vacuolar degeneration,

fatty change and some cells appeared with

pyknotic nuclei (Fig. 5. A).

Collagen content was similar to the

150 mg/kg of cinnamon and control group.

At the electron microscopical level, some

hepatocytes still occupied by fat globules

with decreasing in cytoplasmic organelles

(Fig.5.B).

DISCUSSION

High fructose feeding is commonly

used to induce a strong response on

metabolic parameters. In this study we aimed

to assess the regulatory effect of cinnamon

with different doses on metabolic disorders in

high fructose-fed mice.

Fructose is a highly lipogenic sugar

which is associated with a rapid stimulation

of lipogenesis and an increase of triglycerides

due to its metabolic pathway. In liver, the

most of ingested fructose is metabolized and

converted into glyceraldehyde-3-phosphate

(Tsuchiya et al., 2013).

Numerous studies demonstrated the

association between high fructose diet and

increase of weight gain. El Hasnaoui et al.

(2015) noticed an increase in rat body and

liver weight after feeding high fructose diet

(23%) for 12 weeks. This is not observed in

our study, as body and liver weight data don’t

show any significant changes between treated

groups and control. Consistent with our

results, Tillman et al. (2014) did not record

any significant differences in body weights

between mice fed on high-fructose diet for 14

week and the control.

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We observed impaired glucose

tolerance after the oral glucose load in

fructose-fed mice. Glucose tolerance was

improved in cinnamon at a dose rate 150 mg/

kg group at (t= 60 &120 min) compared with

the control group. Serum blood glucose

levels of the both cinnamon dose groups (150

and 300 mg/kg. b.w) were decreased

compared with the fructose group. Similarly,

Ping et al. (2010) had reported that blood

glucose level decreased after administration

of cinnamon extract at dose of 50 and 100

mg/kg for 35 days in diabetic mice. Yadav et

al. (2007) reported that insulin resistance is

developed after feeding fructose (21%) in

water to rats for 8 weeks. Insulin resistance is

a condition in which circulating insulin

decreases its response to skeletal muscle,

adipose tissues, and liver. The anti-

hyperglycemic effect of cinnamon may be

due to its stimulation of surviving cells to

release more insulin (El Hasnaoui et al.,

2015) and activation of the insulin signaling

(Qin et al., 2004).

In the present study, the measured

hepatic enzymes activities showed no change

among groups. This finding agreed with that

reported by Meeprom et al. (2011). On

contrary, Ismail, (2014) found that

administration of aqueous extract of

cinnamon in dose of 100 and 200 mg/ kg for

6 weeks reduce the elevated ALT and AST in

high fat diet-fed rats.

The current data showed an increase

of total cholesterol, triglycerides, LDL-c, in

the mice fed fructose compared to control

group. These results are consistent with a

previous report (El Hasnaoui et al., 2015).

Administration of cinnamon at both doses to

fructose fed mice resulted in regulation of

lipid metabolism. Consistent with our results,

Kim et al. (2006) and Ping et al. (2010) had

also reported that triglyceride and total

cholesterol levels were decreased, and HDL-

c level was increased in cinnamon treated

mice. This effect may be as a result of

improved insulin action in cinnamon-treated

mice or the direct role of cinnamon on lipid

metabolism by its strong lipolytic activity

(Kannappan et al., 2006).

In the present study, we demonstrated

that serum concentration of TNF-α that were

increased by the high fructose intake was

restored to basal level by cinnamon

supplementation at a dose rate of 150

(mg/kgb.w) not at 300 (mg/kg).There is a

possibility that large doses or chronic

ingestion of cinnamon powder may provide

an increased dose of oil components that may

cause adverse effects, such as inflammation

(Bickers et al., 2005). Abraham et al.,

(2010) mentioned that coumarin, a bioactive

agent present in high level in cinnamon, has

hepatotoxic and carcinogenic effect.

We have observed that the high

fructose intake is associated with increased

oxidative stress as we recorded a depletion of

GSH and CAT enzymes and increase of

MDA level in fructose fed mice when

compared with control. Some authors have

proposed that MDA content increases

significantly in hepatic tissue exposed to high

fructose (Zhang et al. 2013). Excessive

oxidative stress induces mitochondrial

dysfunction, which further disturbs lipid

metabolism and suppress insulin signaling.

On the other hand, the abnormal lipid

metabolism and impaired insulin sensitivity

will enhance oxidative stress (Tsuchiya et

al., 2013). We demonstrated that

administration of cinnamon at the dose of

150 mg/kg significantly increase the hepatic

GSH and CAT contents comparing with

fructose-fed mice. Both doses of cinnamon

were able to decrease MDA level as

compared to fructose-fed mice. This may

attributed to its antioxidant activity due to the

presence of cinnamate (Lee et al., 2003),

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Cinnamaldehyde, phenols and terpenes (Qin

et al., 2010) in the cinnamon bark.

In this study, we observed different

histological alterations in liver sections of

high fructose- fed mice especially in

hepatocytes around central veins as

hepatocellular vacuolation accompanied by

fatty change. That's augmented by electron

microscopy as hepatocyte's cytoplasm

occupied by lipid globules with decreasing in

cytoplasmic organelles. This finding agreed

with that revealed by Schultz et al., (2015),

Prabhakar et al., (2014) and Bradbury and

Berk (2004) who reported that the hepatic

fatty change occurs when the rate or

synthesis of fatty acids by hepatocytes

exceeds the rate of export or catabolism. In

the present study, local and diffuse

inflammatory cells infiltration with von

kupffer cell hyperplasia and congestion of

central veins were recorded. Some authors

(Anstee and Goldin, 2006) presented similar

findings. They stated that increasing in the

inflammatory response occurs as a result of

liver sensitivity for liver fatty change. Some

proliferated collagen fibers were observed

around central vein and periportal hepatic

area as recorded by El Ebiary and Khalaf

(2014). Liver fibrogenesis is occurred in

response to liver injury (Zhao et al. 2012).

So we agreed with the results reported that

fructose was considered as high risk factor in

developing nonalcoholic fatty liver disease

(NAFLD).Our results revealed that co-

administration with cinnamon at dose rate of

150 mg/kg b.w ameliorated the deleterious

effect of fructose on the structure of mice

liver. Most of hepatocytes improved and

appeared as similar to the control group

except some signs of von kupffer hyperplasia

with congested central vein. This

hepatocellular improvement appeared by

electron microscopy which manifested by

disappearance of accumulated lipid droplets,

reappearance of numerous well developed

cytoplasmic organelles. Such results could be

related to the lipolytic, antioxidant and anti-

inflammatory effect of cinnamon (El Ebiary

and Khalaf, 2014 and Yang et al.

2012).While, the liver structure in mice

group treated with 300 mg/kg was not

markedly improved as compared to the lower

dose of cinnamon. Some degree of

degenerations was recognized in our study

with light and electron microscopical

examination. Some of hepatocytes were

vacuolated associated with fatty change and

others had pyknotic nuclei.

In conclusion, the results of this

study revealed that hyperglycemia, lipid

profile disturbance accompanied with

oxidative injury, inflammation and liver fatty

change were induced by high fructose fed in

mice. Co-treatment with cinnamon resulted

in improvement of metabolic disorders and

cinnamon intake at dose of 150 mg/kg was

more effective than the higher dose of 300

mg/kg.

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Fig.2 : A. a photomicrograph of control mice liver showing normal architecture with radiating and

branching hepatic cords from thin walled central vein (CV) separated by blood sinusoids (S) lined with

endothelial cells and von kupffer cells (arrows), polyhedral hepatocytes with granular acidophilic

cytoplasm housing rounded vesicular nuclei. Notice, some cells were binucleated (arrowheads). H&E

stain, 400X. B. showing minimal amount of collagen fibers around central veins (arrow heads) and

periportal hepatic area (arrows). Masson's trichrome stain, 100X. C. An electron micrograph showing

hepatocyte had numerous mitochondria with cristae (m), well developed Golgi apparatus (G), RER (r)

SER (SR). Notice, euchromatic nucleus (N) with prominent nucleoli (n).5800X

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Fig.3: A. a photomicrograph of high fructose fed mice liver showing dilated central vein (CV)

surrounded by affected and degenerated hepatocytes. H&E stain, 100X. B. A higher

magnification of (Fig.2.A) showing vacuolar degeneration of affected hepatocytes with marked

fatty change (arrows). H&E stain, 400X. C. Another section of mice liver showing congested

central veins (CV), diffuse inflammatory cell infiltration (arrows), hepatocytes with cloudy

swelling (double arrows) while others contained pyknotic nuclei (arrowheads). H&E stain,

400X.D. showing apparent increase of collagen fibers around central vein and in the periportal

hepatic area (arrows). Masson's trichrome stain, 100X. E. An electron micrograph showing

accumulation of lipid globules (L) in hepatocyte's cytoplasm, decreasing in cytoplasmic

organelles, housing heterochromatic nucleus (N).1000X.

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Fig.4: A photomicrograph of mice liver fed on fructose and cinnamon 150mg/kg.bw A.showing

great improvement in hepatic architecture as similar to the control group except increasing of

von kupffer cells and congested central veins were present. H&E, 400X. B. showing collagen

fibers returned to be few as the control group. Masson's trichrome stain, 100X. C. An electron

micrograph showing marked improvement with normal hepatocyte's structure. Notice,

euchromatic nucleus (N), numerous mitochondria (m) well developed Golgi apparatus (G) RER

(R) and SER (SR). 7500X.

Fig.5: a photomicrograph of mice liver fed on fructose and cinnamon 300mg/kg.bw showing A.

incomplete improvement compared to the control group, some hepatocytes contained pyknotic

nuclei (arrows) and marked fatty change (arrow heads). H&E, 400X. B. an electron micrograph

showing some hepatocytes accumulated with lipid droplets with few cytoplasmic organelles.

5000X.

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