Honey attenuates the toxic effects of the low dose of tartrazine in male rats

J Food Biochem. 2019;e12780. wileyonlinelibrary.com/journal/jfbc  |_{1 of 11}

https://doi.org/10.1111/jfbc.12780

© 2019 Wiley Periodicals, Inc.

1  | INTRODUCTION

The food additives are substances added to food products to give them a desired food look, taste, better smell, improved texture, and quality (Griffiths & Borzelleca, 2005; Lessof, 2002). Food additives were classified into synthetic or natural. The synthetic

food additives are derived from several sources such as salts and inorganic acids or sulfonated aromatic amines and organic acids, whereas the natural food additives are derived from mineral sources and plants such as fruit juice, chlorophyll, gelatin, caramel color, lipstick tree, beet juice, turmeric yellow, and paprika (Djagny, Wang, & Xu, 2001).

Received: 17 August 2018 

|

  Revised: 2 January 2019 

|

  Accepted: 7 January 2019 DOI: 10.1111/jfbc.12780

F U L L A R T I C L E

Honey attenuates the toxic effects of the low dose of tartrazine in male rats

Haddad A. El Rabey

^1,2

 | Madeha N. Al‐Seeni

³

 | Abdulbasit I. Al‐Sieni

³

 |  Amani Mohammed Al‐Hamed

³

 | Mazin A. Zamzami

^3,4

 | Fahad M. Almutairi

¹

Abbreviations: ALP, alkaline phosphatase; ALT, alanine aminotransferase; AST, aspartate aminotransferase; BHT, butylated hydroxytoluene; BW, body weight; BWG, body weight gain;

Cat, catalase; FER, food efficiency ratio; G1, the first negative control untreated group fed basal diet containing 3.75 mg/kg b.w. sulfanilic acid; G2, The second positive control group re‐

ceived diet containing 10 mg/kg b.w. tartrazine and 3.75 mg/kg b.w. sulfanilic acid; G3, The third group (G3) received 10 mg/kg b.w. tartrazine and 3.75 mg/kg b.w. sulfanilic acid and cotreated with a daily dose of 2.5g/ Kg body weight of 25% aqueous solution of honey by gastric pipette for 8 weeks; GR, Glutathione reductase; GSH, Glutathione reduced; LDL, low‐

density lipoprotein; N.N cellulose, nonnutritive cellulose; SOD, Super oxide dismutase; TC, total cholesterol; TG, triglyceride; VLDL, very‐low‐density lipoproteins.

1Biochemistry Department, Faculty of Science, University of Tabuk, Tabuk, KSA

2Bioinformatics Department, Genetic Engineering and Biotechnology Research Institute, University of Sadat City, Sadat City, Egypt

3Biochemistry Department, Faculty of Science, King Abdulaziz University, Jeddah, KSA

4Cancer and Mutagenesis Unit, King Fahd Center for Medical Research, King Abdulaziz University, Jeddah, KSA

Correspondence

Haddad A. El Rabey, Biochemistry Department, Faculty of Science, University of Tabuk, Tabuk, KSA.

Email: helrabey@ut.edu.sa Funding information

King Abdulaziz University, Grant/Award Number: (270 أط‐35‐).

Abstract

Honey is traditionally used in burns, wound healing, ulcers, boils, and fistulas. Honey was tested to prevent tartrazine toxicity in male rats for 8 weeks. The 18 rats of the experiment were randomly divided into three 6‐rat groups. The negative control group (G1) fed diet with sulfanilic acid, the tartrazine positive group (G2) fed diet containing tartrazine and sulfanilic acid and the honey‐treated group (G3) fed diet as in G2 and cotreated with honey. Tartrazine decreased antioxidants, high‐density lipo‐

proteins and proteins, and increased liver enzymes, kidney indices, lipid peroxidation, triglycerides, total cholesterol, and low‐ and very‐low‐density lipoproteins. In addi‐

tion, tartrazine‐treated group showed drastic damage of the tissues of stomach, liver, kidney, and testis. Honey treatment increased antioxidants and high‐density lipopro‐

teins, and decreased lipid peroxidation, liver enzyme and kidney parameters. Honey treatment also improved stomach, liver, kidney, and testis tissues. In conclusion, honey protects male rats against tartrazine toxicity.

Practical applications

Honey was tested to prevent tartrazine toxicity in male rats in an experiment con‐

ducted for 8 weeks. Catalase, glutathione reductase, superoxide dismutase, glu‐

tathione reduced, the low‐ and high‐density lipoproteins, lipid peroxidation, liver enzyme, and kidney parameters were measured to evaluate both the toxic effect of tartrazine in G2 and the protective potential of honey in G3.

K E Y W O R D S

azo dye, honey, kidney, liver, rat, stomach, tartrazine

The azo dye tartrazine is widely used additive in food coloring, food products, and drugs (Walton et al., 1999). It is the fourth most commonly used coloring additive (Pollock et al., 1989). Excess of tartrazine can cause adverse effects on health (Al‐Seeni, Rabey, Al‐

Hamed, & Zamazami, 2018). Food additives that contain butylated hydroxytoluene (BHT) such as tartrazine, allura red, erythrosine, and indigo carmine induce toxic effect on liver (Grunow, 1999; Safer &

AI‐Nughamish, 1999). The high dose (500 or 1,000 mg/kg) of BHT causes centrilobular necrosis in liver, elevated serum transaminase activities, and hemorrhagic death, after 24 hr of administration (Nakagawa, Tayama, Nakao, & Hiraga, 1984). It also increases total blood cholesterol, the weight of pancreas, liver and mammary glands, and induces liver and uterus tumors (Safer & AI‐Nughamish, 1999).

Chequer, Dorta, and Oliveira (2011) stated that the toxicity of tartrazine results from the metabolic reduction of the azo bond.

In addition, the azo dyes are oxidized by P450 enzymes to N‐hy‐

droxy derivatives after the complete reduction into aromatic amines (Umbuzeiro et al., 2005).

Honey is a natural product with a high nutritious value. It is com‐

posed primarily of glucose, fructose, and 4 to 5% fructo‐oligosac‐

charides which is a prebiotic agents (Al‐Seeni, Rabey, & Al‐Solamy, 2015). In addition to its use as food, honey is considered one of the oldest known medicines (Jones, 2001). It acts as an antioxidant, anti‐

inflammatory, and anti‐tumor. Moreover, it normalizes kidney func‐

tions and protects the liver from intoxication (Al‐Seeni et al., 2015;

Bariliak, Berdyshev, & Dugan, 1996).

In the present study, honey was tested to prevent tartrazine tox‐

icity in male rats.

2  | MATERIALS AND METHODS

2.1 | Materials and diet

The Sigma brand tartrazine was purchased from Sigma (USA), whereas the natural Sidr bee honey (from Yemen) was purchased from a honey shop (Jeddah, Saudi Arabia).

Parameters Statistics

G1 Negative control

G2 Positive

control G3 Honey

ALT (U/L) Mean ± SE 27.83 ± 1.47^c 77.16 ± 1.68^a 51.33 ± 1.20^b LSD 0.05 = 4.054

T‐test – −30.33*** 10.08***

AST (U/L) Mean ± SE 30.50 ± 1.11^c 79.16 ± 1.57^a 55.50 ± 1.45^b LSD 0.05 = 4.054

T‐test – −30.71*** 9.04***

ALP (U/L) Mean ± SE 158.00 ± 3.89^c 275.83 ± 3.56^a 199.33 ± 4.50^b LSD 0.05 = 12.643

T‐test – −24.62*** 12.61***

Total protein

(g/dl) Mean ± SE 7.54 ± 0.05^a 5.39 ± 0.20^c 6.58 ± 0.07^b

LSD 0.05 = 0.354

T‐test – 10.25*** ***−5.28

Albumin (g/

dl)

Mean ± SE 4.28 ± 0.09^a 2.65 ± 0.14^c 3.63 ± 0.06^b LSD 0.05 = 0.265

T‐test – 12.73*** ***−7.27

Globulin (g/L) Mean ± SE 3.25 ± 0.12^a 2.74 ± 0.19^a 2.95 ± 0.10^a LSD 0.05 = 0.582

T‐test – 2.33* −0.81^NS

A/G ratio (g/L)

Mean ± SE 1.32 ± 0.07^a 0.99 ± 0.11^b 1.23 ± 0.06^b LSD 0.05 = 0.251

T‐test – 3.28* −1.81^NS

Total Bilirubin (mg/dl)

Mean ± SE 0.48 ± 0.02^b 0.70 ± 0.04^a 0.53 ± 0.01^b LSD 0.05 = 0.084

T‐test – −4.98*** 3.31**

Note. ANOVA analysis: within each row, means with different superscript (a, b, c, or d) are signifi‐

cantly different at p < 0.05, whereas means superscripts with the same letters mean that there is no significant difference at p > 0.05. LSD: least significant difference. NS: Nonsignificant. Data are rep‐

resented as mean ± SE. T‐test values;

***significant at p < 0.001.

TA B L E 1  Effect of honey

supplementation for 8 weeks on serum liver enzymes, total protein and bilirubin in rats cosupplemented with tartrazine

Each 100 g basal diet consisted of the following ingredients (grain mill, Jeddah, Saudi Arabia): 100 g diet were mixed: 12% pro‐

tein (17.14 g 70% casein), 4 g minerals (4% minerals), 4 g cellulose

(4% fiber), 4 g corn oil (4% fat), 1 g vitamin mixture (1% vitamin), 0.2 g choline chloride (0.2%), 0.3 g methionine (0.3%), and 69.36 g corn starch (69.36%).

Parameters Statistics G1‐ve Control

G2 Positive

Control G3 Honey

S.TC (mg%) Mean ± SE 164.166 ± 2.25^c 261.50 ± 3.12^a 203.17 ± 4.67^b LSD 0.05 = 9.945

T‐test – −61.42*** 11.18***

S.T.G (mg/dl) Mean ± SE 133.17 ± 2.71^c 224.67 ± 3.25^a 195.17 ± 2.78^b LSD 0.05 = 8.793

T‐test – −51.28*** 6.15***

S.HDLc (mg/dl) Mean ± SE 46.50 ± 0.95^c 32.50 ± 0.76^a 38.66 ± 0.55^b LSD 0.05 = 2.246

T‐test – 8.79*** −5.72***

S.LDLc (mg/dl) Mean ± SE 89.83 ± 1.95^c 183.83 ± 3.04^a 125.67 ± 4.46^b LSD 0.05 = 9.218

T‐test – −19.05*** 11.80***

V.LDLc (mg/dl) Mean ± SE 26.63 ± 0.54^c 44.93 ± 0.65^a 39.03 ± 0.55^b LSD 0.05 = 1.750

T‐test – −9.28*** 6.15***

Note. ANOVA analysis: within each row, means with different superscript (a, b, c) are significantly different at p < 0.05, whereas means superscripts with the same letters mean that there is no signifi‐

cant difference at p > 0.05. LSD: least significant difference. Data are represented as mean ± SE. T‐

test values;

***significant at p < 0.001.

TA B L E 2  Effect of honey

supplementation for 8 weeks on serum lipid profile in rats cosupplemented with tartrazine

TA B L E 3  Effect of honey supplementation for 8 weeks on kidney functions and electrolytes in rats cosupplemented with tartrazine

Parameters Statistics G1 Negative control G2 Positive control G3 Honey

Uric acid (mg/dl) Mean ± SE 4.18 ± 0.06^c 6.70 ± 0.09^a 5.71 ± 0.13^b

LSD 0.05 = 0.2766

T‐test – −27.66*** 7.55***

Creatinine (mg/dl) Mean ± SE 0.66 ± 0.03^c 2.91 ± 0.11^a 1.66 ± 0.07^b

LSD 0.05 = 0.270

T‐test – −19.13*** 11.82***

Urea (mg/dl) Mean ± SE 23.76 ± 0.88^c 65.66 ± 1.42^a 58.50 ± 1.82^b

LSD 0.05 = 4.425

T‐test – −19.84*** 3.30**

Na⁺ (mmol/l) Mean ± SE 139.33 ± 1.52^a 140.83 ± 1.01^a 140.33 ± 1.56^a

LSD 0.05 = 3.878

T‐test – −1.01^NS 0.27^NS

K⁺ (mmol/l) Mean ± SE 4.73 ± 0.06^a 4.65 ± 0.07^a 4.71 ± 0.06^a

LSD 0.05 = 0.211

T‐test – 0.75^NS −2.00^NS

Note. ANOVA analysis: within each row, means with different superscript (a, b, c) are significantly different at p < 0.05, whereas means superscripts with the same letters mean that there is no significant difference at p > 0.05. LSD: least significant difference. NS: Nonsignificant. Data are represented as mean ± SE. T‐test values;

**significant at p < 0.01, ***significant at p < 0.001.

2.2 | Animal housing and experiment design

Eighteen male rats (Rattus norvegicus, East China Origin) weigh‐

ing 175–185 g were purchased from The Faculty of Pharmacy, King Abdulaziz University, Jeddah, KSA. Animal experiments were achieved under an approved protocols (The Animal House, University of King

Abdulaziz, Jeddah, Kingdom of Saudi Arabia). Animals were housed six/polycarbonate cages. Water bottles cages and bedding were re‐

placed twice a week. Before the beginning of the experiment, the rats were kept under observation for 2 weeks to avoid any undercurrent infection. Rats were divided into three 6‐rat groups. The negative control group (G1) fed basal diet containing 3.75 mg/kg b.w. sulfanilic TA B L E 4  Effect of honey supplementation for 8 weeks on antioxidants in serum, liver, and kidney tissue homogenate of rats

cosupplemented with tartrazine

Parameters Serum Statistics G1 Negative control G2 Positive control G3 Honey

Serum Catalase (CAT) (U/ml) Mean ± SE 0.96 ± 0.03^a 0.19 ± 0.01^c 0.61 ± 0.02^b

LSD 0.05 = 0.059

T‐test – 25.30*** −15.23***

Superoxide dismutase (SOD) (U/ml)

Mean ± SE 544.32 ± 11.59^a 131.67 ± 1.24^c 394.23 ± 5.74^b LSD 0.05 = 22.014

T‐test – 32.68*** −41.54***

Glutathione reduced

(GSH) (mmol/ml) Mean ± SE 4,762.20 ± 25.60^a 2,369.00 ± 15.09^c 3,531.70 ± 57.36^b LSD 0.05 = 126.857

T‐test – 75.84*** −19.10***

Glutathione reduc‐

tase (GR) (U/ml)

Mean ± SE 5.80 ± 0.13^a 1.40 ± 0.06^c 4.08 ± 0.13^b

LSD 0.05 = 0.342

T‐test – 34.31*** −16.42***

Liver Catalase (CAT) (U/g) Mean ± SE 5.53 ± 0.09^a 1.88 ± 0.07^c 3.11 ± 0.16^b

LSD 0.05 = 0.356

T‐test – 41.24*** −7.78***

Superoxide dismutase (SOD) (U/g)

Mean ± SE 920.50 ± 5.20^a 340.67 ± 5.33^c 737.00 ± 4.91^b LSD 0.05 = 14.695

T‐test – 172.53*** −65.17***

Glutathione reduced (GSH) (mmol/g)

Mean ± SE 7,599.70 ± 7.59^a 3,196.20 ± 4.96^c 5,359.20 ± 5.07^b LSD 0.05 = 22.157

T‐test – 395.36*** −300.33***

Glutathione reductase (GR) (U/g)

Mean ± SE 9.15 ± 0.20^a 2.30 ± 0.11^c 6.56 ± 0.08^b

LSD 0.05 = 0.428

T‐test – 24.55*** −31.41***

Kidney Catalase (CAT) U/g. Mean ± SE 4.60 ± 0.09^a 1.36 ± 0.08^c 2.46 ± 0.12^b

LSD 0.05 = 0.317

T‐test – 4.71*** −9.08***

Superoxide dismutase(SOD) (U/g)

Mean ± SE 762.83 ± 4.96^a 261.17 ± 1.92^c 527.50 ± 3.85^b LSD 0.05 = 10.867

T‐test – 96.23*** −56.89***

Glutathione reduced(GSH) (mmol/g)

Mean ± SE 7,198.80 ± 7.40^a 2,749.30 ± 14.51^c 3,196.20 ± 0.96^b LSD 0.05 = 31.468

T‐test – 353.33*** 92.73***

Glutathione reduc‐

tase (GR) (U/g)

Mean ± SE 7.28 ± 0.11^a 1.95 ± 0.11^c 5.31 ± 0.10^b

LSD 0.05 = 0.317

T‐test – 27.44*** −38.17***

Note. ANOVA analysis: within each row, means with different superscript (a, b, c) are significantly different at p < 0.05, whereas means superscripts with the same letters mean that there is no significant difference at p > 0.05. LSD: least significant difference. Data are represented as mean ± SE. T‐

test values;

***significant at p < 0.001.

acid. The tartrazine positive group (G2) fed a diet containing 3.75 mg/

kg b.w. sulfanilic acid and 10 mg/kg b.w. tartrazine. The honey‐treated group (G3) fed the same dose of sulfanilic acid and tartrazine as in G2 and simultaneously cotreated with a daily dose of 25% aqueous solu‐

tion of honey (2.5 g/Kg body weight) by oral gavage (Yamada, Itoh,

& Murakami, 1999). The experiment was conducted for 8 weeks. The test diet was prepared by mixing the required amount of tartrazine that dissolved in distilled water with sulfanilic acid, with the diet. Feed and water were available ad libitum. At the end of the experiment, necropsy was done to all rats.

2.3 | Tissue homogenate

The tissue homogenate for liver and kidney was prepared according to Al‐Malki and El Rabey (2015).

2.4 | Blood collection

At the end of the experiment, blood samples were collected from the dorsal pedal vein in plain tubes for biochemical analyses. Blood sam‐

ples were centrifuged at room temperature (for 10 m at 1,000 rpm), and then analyzed.

TA B L E 6  Effect of honey supplementation for 8 weeks on body weight gain (BWG) and food efficiency ratio (FER) in rats cosupplemented with tartrazine

Physiological evaluation parameters Statistics G1 Negative control G2 Positive control G3 Bee honey

BWG g /8 week Mean ± SE 46.00 ± 1.064^a −39.50 ± 1.727^d 22.16 ± 1.720^c

LSD 0.05 = 4.816

T‐test – 44.75*** −25.36***

BWG g/day Mean ± SE 0.763 ± 0.017^a 0.655 ± 0.029^a 0.366 ± 0.028^c

LSD 0.05 = 0.078

T‐test – 2.92** 7.15***

BWG % Mean ± SE 25.29 ± 0.606^a 13.57 ± 6.977^d 10.56 ± 0.830^c

LSD 0.05 = 2.400

T‐test – 5.94*** −3.14**

FER g/day Mean ± SE 0.041 ± 0.000^a −0.011 ± 0.015^b 0.020 ± 0.001^a

LSD 0.05 = 0.084

T‐test – 3.58** −1.93^NS

FER % Mean ± SE 4.116 ± 0.094^a 3.585 ± 0.152^d 2.000 ± 0.167^c

LSD 0.05 = 0.458

T‐test – 45.12*** −23.44***

Note. ANOVA analysis: within each row, means with different superscript (a, b, c, or d) are significantly different at p < 0.05, whereas means super‐

scripts with the same letters mean that there is no significant difference at p > 0.05. LSD: least significant difference, NS: Nonsignificant. Data are represented as mean ± SE. T‐test values;

**significant at p < 0.01, ***significant at p < 0.001.

Parameters Statistics

G1 Positive control

G2 Negative

control G3 Honey

MDA Mean ± SE 1.50 ± 0.09^c 3.50 ± 0.10^a 2.53 ± 0.16^b

nmol/ml LSD 0.05 = 0.394

Serum T‐test – −10.64*** 5.15***

MDA Mean ± SE 1.81 ± 0.12^c 11.06 ± 0.23^a 6.86 ± 0.15^b

nmol/g. LSD 0.05 = 0.535

Liver tissue T‐test – −33.44*** 12.22***

MDA nmol/g. Mean ± SE 1.08 ± 0.08^c 7.83 ± 0.16^a 4.53 ± 0.11^b Kidney tissue LSD 0.05 = 0.402

In kidney tissue nmol/g. Kidney tissue

T‐test – −28.25*** 27.24***

Note. ANOVA analysis: within each row, means with different superscript (a, b, c) are significantly different at p < 0.05, whereas means superscripts with the same letters mean that there is no signifi‐

cant difference at p > 0.05. LSD: least significant difference. Data are represented as mean ± SE. T‐

test values;

***significant at p < 0.001.

TA B L E 5  Effect of honey supplementation for 8 weeks on lipid peroxide in rats cosupplemented with tartrazine

After slaughtering the rats, the kidney, the liver, testes, and the stomach were dissected out and washed in sterile saline. A kid‐

ney, a piece of the liver, stomach, and a testis were fixed in 10%

buffered formalin for histopathological studies. A piece of the liver and the other kidney were kept ice‐cold for tissue homogenate preparation.

2.5 | Liver function enzymes

Human kits (Germany) were used in estimating the activity of alanine aminotransferase (ALT), alkaline phosphatase (ALP), and aspartate aminotransferase according to the instruction of the supplier.

2.6 | Serum lipids

The method of Naito (1984) was used in estimating the total choles‐

terol (TC) and the triglycerides (TG) in the serum using Human Kit (Germany). Whereas the method described by Gordon, Castelli, and Hjortland (1977) was used in estimating the high‐density lipoprotein (HDL) using Human kit (Germany). The low‐density lipoprotein (LDL) was estimated in the serum according to the equation of Friedewald,

Levy, and Fredrickson (1972). All tests were carried out according to the instruction of the supplier.

2.7 | Serum bilirubin

The method of Balistreri and Shaw (1987) was used in estimating total serum bilirubin using spectrum kit (Germany).

2.8 | Total protein

The method of Weissman, Schoenbach, and Armstead (1950) was used in estimating total serum proteins using Human Kit (Germany), whereas the method of Rebecca (2006) was used in estimating serum albumin using Sigma‐Aldrich kit (USA).

2.9 | Kidney indices and serum electrolytes

The method of Fawcett and Scott (1960) was used in estimating serum creatinine, urea, and uric acid using Human kit (Germany), whereas the method of Berry, Mazzachi, Pejakovic, and Peake (1988) was used in estimating sodium and potassium ions (Na⁺, K⁺, respec‐

tively) using Human kit (Germany).

F I G U R E 1  (a) Normal renal tissues of control group (G1), (b) Renal tissues of G2 that was treated showing an enlargement of renal glomeruli with vacuolations, (c) nearly normal renal tissues of the honey treated group (G3), short arrow: uriniferous tubules, long arrow:

glomerulus (X 200, H&E stains)

2.10 | Estimation of antioxidants and lipid peroxidation

The method of Aebi (1984) was used in estimating catalase (Cat), super oxide dismutase (SOD), glutathione reductase (GR), Glutathione re‐

duced (GSH), and lipid peroxidation using Biodiagnostic kit (Egypt).

2.11 | Histopathology

Microscopic sections of kidney, liver, testis, and stomach were pre‐

pared according to the method of Drury, Wallington, and Cancerson (1976). An Olympus light microscope was used for examining and photographing the sections.

2.12 | Statistical analysis

The SPSS program was used in analyzing the resulted data. The test of sig‐

nificance (t‐test) and the mean ±SD was calculated, whereas the SAS pack‐

age was used in calculating the one‐way analysis of variance (ANOVA, p < 0.05) using a protected least significant difference (LSD) test.

2.13 | Physiological evaluation

The body weight gain (BWG) and its percentage (BWG%), the food efficiency ratio (FER) and its percentage (FER%) were calculated ac‐

cording to Davies and Morris (1993).

3  | RESULTS

3.1 | Liver Function, Bilirubin, and Total Proteins

Tartrazine administration in G2 significantly increased AST, ALP, and ALT in the serum compared to G1 (Table 1). In G3, the cotreatment with honey nearly restored ALT, AST, and ALP to the normal val‐

ues. In contrast, tartrazine toxicity in G2, decreased serum albumin and globulin, and albumin/globulin ratio, whereas increased bilirubin compared to G1. The cotreatment with honey in G3 nearly restored the total protein and bilirubin to the normal.

3.2 | Lipid Profile

Tartrazine administration in G2 significantly increased TG, TC, LDL, and VLDL, whereas decreased HDL compared to G1 (Table 2). While, the cotreatment with honey in the third group (G3) significantly de‐

creased TG, TC, LDL, and VLDL and increased HDL compared to the positive control group (G2).

3.3 | Kidney Function and Electrolytes

Tartrazine administration significantly increased the mean values of kidney function parameters (urea, creatinine, and uric acid) and serum electrolytes (Na⁺ and K⁺) in G2 compared to that of the neg‐

ative control in G1 (Table 3). In G3, honey treatment decreased the

F I G U R E 2  (a) Normal hepatic tissues of G1, (b) hepatic tissues treated with tartrazine (G2), (c) nearly normal hepatic tissues of the honey treated group (G3), arrows: hepatocytes (X 200, H&E stains)

kidney indices, serum Na⁺ and K⁺ compared to G2 and restored them nearly to the values of the negative control.

3.4 | Antioxidants

Tartrazine administration in G2 decreased SOD, Cat, GR, and GSH in serum, liver, and kidney tissue homogenate compared to G1 (Table 4).

The cotreatment with honey in G3 significantly decreased all these antioxidants in the serum, liver, and kidney tissue homogenate and restored them nearly to the physiological values.

3.5 | Lipid Peroxidation

Tartrazine administration significantly increased lipid peroxidation in the serum, liver, and kidney tissue homogenate of G2 as shown in Table (5).

Treating G3 rats with honey significantly decreased lipid peroxidation compared to G2 and restored it nearly to the normal values as in G1.

3.6 | Physiological Evaluation

Tartrazine administration in G2 significantly decreased BWG (g/day, and 8 weeks), BWG%, FER and FER% compared to G1 (Table 6). In G3, treating with honey significantly ameliorated these parameters compared to G2.

3.7 | Histopathology

The normal renal tissues of G1 are shown in Figure 1a. Figure 1b shows renal tissues of G2 showing an enlargement of the renal glo‐

meruli with vacuolations and hypertrophy and degeneration of the lining epithelial cells of the renal tubules. Figure 1c shows renal tis‐

sues of “G3” restored nearly their normal histology.

The normal hepatic tissues of G1 are shown in Figure 2a.

Figure 2b shows the hepatic tissues of G2 with necrosis, congestion of portal vein with erythrocytes and some lymphocytes as a result of tartrazine toxicity. The cotreatment with honey in G3 (Figure 2c) nearly restored hepatic tissues and hepatic strands to the normal state.

The normal testicular tissues of G1 with normal seminiphrous tu‐

bules and normal spermatogenesis are shown in Figure 3a, whereas an arrested spermatogenesis at different levels of G2 are shown in Figure 3b. The cotreatment with honey in G3 shows nearly normal seminipherous tubules and spermatogenesis (Figure 3c).

The normal stomach tissues of G1 shows normal tunica mucosa and intact lining epithelium (Figure 4a) shows. Figure 4b shows that tartrazine toxicity in G2 caused degenerated tunica mucosa and lining epithelium. On the other hand, Figure 4c shows the stomach tissue of G3 with nearly restored normal gastric mucosa and lining epithelium.

F I G U R E 3  (a) Normal testicular tissues of G1, (b) Arrested testicular tissues of (G2), (c) nearly normal testicular tissues of the honey treated group (G3), arrow: seminipherous tubules and spermatogenesis (X 200, H&E stains)

4  | DISCUSSION

The natural constituent of honey makes it one of the common used natural product reservoir for treating many diseases. In the cur‐

rent study, honey was tested in attenuating and protecting against the adverse effects of tartrazine toxicity in male rats. The increase in TG, TC, LDL, and VLDL, and the decrease in HDL in the tartra‐

zine positive group (G2) compared to the negative control group (G1) is consistent with Aboel‐Zahab et al. (1979). The cotreatment with honey in G3 ameliorated the lipid profile by decreasing TG, TC, LDL, and VLDL, and increasing HDL. This may be due to the scavenging reducing power of monosaccharaides constituting honey (Hassan, 2007).

Similarly, the increase in liver and kidney functions parameters as a result of tartrazine toxicity agrees with previous studies (Al‐Seeni et al., 2018; Amin, Abdel Hameid, & Abd Elsttar, 2010; Ashour &

Abdelaziz, 2009) and restored nearly to normal with honey treat‐

ment. The improvement in liver and kidney function after treatment with honey supports the fact that the antioxidant content of honey award it a protective activity to both liver and kidney (Al‐Seeni et al., 2015; Galal, Zaki, & Seif El‐Nasr, 2012; Omotayo, Sulaiman, &

Wahab, 2012; El Rabey, Al‐Seeni, & Al‐Solamy, 2013).

The increase in total protein and the decrease in bilirubin in G3 compared to the positive control group as a result of honey treat‐

ment is consistent with Hassan (2007), Afroz et al. (2014) and Al‐

Seeni et al. (2015). This amelioration with honey treatment is due to its antioxidant activity and its ability to reduce reactive oxygen intermediates that also decrease bilirubin in male rats (Al‐Seeni et al., 2015; Al‐Seeni, Rabey, & Zamzami, 2016).

The decrease in CAT, SOD, GSH, and GR in the serum, kidney, and liver tissue homogenate of G2 compared to the negative control (G1) as a result of tartrazine toxicity is consistent with Gao et al.

(2011). Whereas the increase in these antioxidants in the serum, and kidney and liver tissue homogenate compared to the positive con‐

trol as a result of honey treatment in G3 is consistent with Hassan (2007), Afroz et al. (2014) and Al‐Seeni et al. (2016).

In contrast, the increase in lipid peroxidation in the serum, liver, and kidney tissue homogenate of the positive control group as a re‐

sult of tartrazine toxicity, and its decrease with honey treatment is consistent with other studies (Hassan, 2007, Afroz et al., 2014 and Al‐Seeni et al., 2016).

The decrease in BWG and BWG%, FER and FER% as a result of tartrazine administration is in agreement with Mehedi et al. (2013).

Honey treatment restored the BWG nearly to the normal values. The F I G U R E 4  (a) Normal lining tissues of (G1), (b) degenerated lining epithelium of stomach of G2, (c) nearly normal stomach lining tissues of the honey treated group (G3), arrow: lining cells (X 200, H&E stains)

increase in FER and FER% with honey treatment is consistent with Hassan (2007).

The drastic pathological changes in tissues of the studied organs (kidney, liver, testis, and stomach) of the tartrazine‐treated group is consistent with Mehedi et al. (2013) and Al‐Seeni et al. (2018).

Restoring the studied tissues their normal histology with honey treatment may be due to its content of ascorbic acid, flavonoids, catalase, phenolic compounds, and tocopherols (Abdel‐Moneim &

Ghafeer, 2007; Al‐Seeni et al., 2015; Hassan, 2007; Sarkar & Ghosh, 2012).

In conclusion, honey succeeded in protecting against tartra‐

zine toxicity by restoring the biochemical and histopathological changes. It ameliorated the biochemical parameters and restored the histology of kidney, liver, testis, and stomach nearly to the nor‐

mal values.

ACKNOWLEDGMENTS

This project was funded by King Abdulaziz University, Jeddah, under grant No. (270 35‐طأ‐). The authors, therefore, acknowledge with thank DSR technical and financial support.

CONFLIC T OF INTEREST

The authors of this paper have no conflict of interests.

ORCID

Haddad A. El Rabey https://orcid.org/0000‐0002‐4347‐6864

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How to cite this article: El Rabey HA, Al‐Seeni MN, Al‐Sieni AI, Al‐Hamed AM, Zamzami MA, Almutairi FM. Honey attenuates the toxic effects of the low dose of tartrazine in male rats. J Food Biochem. 2019;e12780. https://doi.

org/10.1111/jfbc.12780