Effect of Grape seed extract in blood parameters associated with Experimental Thyroid Disorders in White Male Rats

 

Jameel K. Alumeri, Ahmed Jassem AL-Naely

 

ABSTRACT:

 

 

INTRODUCTION:

Thyroid hormones disorders consider as common health problem in 1% of general human and 5% of people over age 60 years as thyroid hormones play many physiological roles in the human body most important one is regulate hematopoiesis in bon marrow¹.

 

The relationship between thyroid hormone disorders and blood parameters has been known from ancient times, In 1979 the researcher Fein mentioned that Graves' disease association with anemia as well as² noted there is decrease in red blood cells after thyroidectomy in peripheral blood, on the other hand hypothyroidism can cause form of anemia usually is macrocytic hypochromic anemia in moderate severity while hyperthyroidism commonly cause erythrocytosis³. hypothyroidism and hypothyroidism cause a state of oxidative stress and this in turn affects the nature of plasma lipid^4,5. In general many studies have found that blood parameters are normal if euthyroid stat is achieved⁶.

 

Carbimazole is anti-hyperthyroidism drug by reducing the intake of iodine and formation of diodotyrosine which prevents coupling of thyroid peroxidase enzyme and iodination of tyrosine residues on thyroglobulin and reduce production of thyroid hormones lead hypothyroidism⁷, L-thyroxin is anti-hypothyroidism drug can be used to induced hyperthyroidism in rodents⁸.

 

Grape seed extract (GSE) is a nutritional supplement with important health benefits^9, Because it is a complex mixture of polyphenols consist of flavonoids and nonflavonoids compounds¹⁰. This plant extract has many benefits include cardioprotective benefits¹¹, Renoprotective¹² as well as neuroprotective¹³. The grape seed extract GSE known for its ability to treat anemia and has various biological functions due to it rich polyphenol ingredients¹⁴, Therefore, it was used in this study to verify its role in reducing the effects of thyroid disorders on some blood parameters.

 

MATERIAL AND METHODS:

Sixty healthy male rats 210-250g weight and 10-12 weeks age were used in the current study, divided in six groups (n=10) each group Treated as flow: control group (C) given distil water for 45 days. First treatment (T1) given oral dose of 30mg/kg carbimazole for 45 days as control positive, Second treatment (T2) given oral dose of 20mg/kg L-thyroxin for 45 days as control positive, Third treatment (T3) given oral dose of 150mg/kg GSE for 45 days as control negative, Fourth treatment (T4) given oral dose of 30mg/kg carbimazole for 15 days to induced hypothyroidism then given 30mg/kg carbimazole synchronously with 150mg/kg GSE for 30 days. Fifth treatment (T5) given oral dose of 20mg/kg L-thyroxin for 15 days to induced hyperthyroidism then given 20mg/kg L-thyroxin synchronously with 150 mg/kg GSE for 30 days.

 

Plant extract: Local Iraqi Grape was purchased from local Iraqi markets, Seeds were isolated and drying it in the shade, then grind by an electric mixer for use in the Soxhlet apparatus for water extract of grape seeds.

 

Animals scarified: In 46 day animals were sacrificed blood samples were collected by directly heart puncture for blood parameters assays by hematology analyzer apparatus model.

Statistical Analysis

 

All results of the study were subjected for statistical analysis in order to know the significant differences between the groups by using F-test at 0.05 probability level¹⁵.

 

RESULTS:

The results (table1) showed that there was no significant decrease (P>0.05) in RBC result in T1 but was there significant increase (P<0.05) in T2 treatment compared with control, while results showed significant decrease (P<0.05) in each of PCV, Hb and ESR in booth T1 and T2 treatments compared with control, Total count of WBC results(table 2) showed significant increase (P<0.05) in T1 and significant decrease (P<0.05) in T2 compared with control, This was accompanied by significant differences in MO, ECIDO, LY and NE when comparing the results of these parameters in T1 and T2 treatments with control.

 

As for T3 treatment which in turn were closely related without no significant difference with control group.

 

Fourth group results experienced improvement in RBCs and Hb closely to control whereas PCV parameter was relative to control and also not significantly different from its rate in the first group, ESR results in this group did not differ significantly compared with control group while WBC results decreased significantly (P<0.05) compared with control and T1. The fifth group result they were close to normal range of all parameters in control group as well as compared with third treatment, in contrast results of this fifth treatment were differ significantly towards improvement in these parameters compared with second group which treated with L-thyroxin alone.

 

Table (1): Grape seeds extract effects in some blood parameters in white male rat.

Parameters   Treatments	RBCs (x1012/L)	PCV %	Hb g/dl	ESR
Control	7.45±0.13 b	45.82±2.12 a	13.65±0.66 a	4.49±0.33 a
T1	7.54±0.39 b	40.36±3.17 c	11.70±1.03 c	3.33±0.33 b
T2	8.23±0.26 a	43.33±1.73 b	10.96±0.52 b	3.00±0.57 b
T3	7.39±0.11 b	45.43±0.22 a	13.77±0.54 a	4.79±0.23 a
T4	7.76±0.45 b	41.40±1.53 ac	12.56±0.43 a	4.33±0.11 a
T5	8.01±0.20 ab	44.96±0.75 a	13.53±0.26 a	4.00±0.02  a

Means ± SE

 

Table (2): Grape seeds extract effects in total and deferential WBC count.

Parameters Treatments	WBCs (x109/L)	NE %	LY %	MO %	ECIDO %
Control	12.195±0.37 b	27.13±0.33 a	69.58±0.09 c	2.29±0.33 a	1.00±0.001 a
T1	15.51±2.46 a	29.43±4.83 a	69.40±5.57 c	0.80±0.56 b	0.37±0.006 b
T2	6.35±2.93 c	14.26±3.45 b	85.53±9.45 a	0.1667±0.33 a	0.0667±0.006 c
T3	13.14±0.55 b	29.39±0.52 a	67.61±2.22 c	2.00±0.57 a	1.00± 0.001 a
T4	12.99±2.33 b	25.71±5.03 a	72.76±6.11 b	1.20±0.001 b	0.333±0.001 b
T5	7.91±0.44 c	25.46±4.02 a	72.40±4.18 b	0.400±0.30 a	0.0333±0.003 d

Means ± SE

 

DISCUSSION:

Thyroid hormones regulates body metabolism so any functional or histological disorder in thyroid gland maybe effects functions of cells and organs of the body this includes bone marrow which affected by thyroid disorders this causes oxidative stress especially in hypothyroidism leading to anemia^16,17, In our result there was no significant decrease in RBCs count in carbimazole compared with control treatment but we noticed differ form of anemia represent by significant decrease in Hb and PCV compared with control that maybe refer to microcytic hypochromic anemia, Many studies pointed out that hypothyroidism cause various forms of anemia like microcytic hypochromic anemia this generally attributed to malabsorption of iron¹⁸ Because of most of previous studies have confirmed a relationship between hypothyroidism and iron deficiency ¹⁹.

 

Microcytic hypochromic anemia means small size and low weight of erythrocytes due to lack of quantity of hemoglobin this can cause delay red blood cells to fall into the test tube and thus maybe the reason for decrease the erythrocyte sedimentation rate as we not in carbimazole treatment. Significant increase in RBC count in L-thyroxin treatment may be due to increase the production of red blood cells from the bone marrow, because of in hyperthyroidism erythrocytosis is common ³ by two way either increase secretion of erythropoietin from kidney which is stimulates erythrocytes production from bone marrow^{7,8 ,} or by increase basal metabolic rate through thyroid hormones which accelerate RBC formation²⁰, This rapid production process consider as a kind of stress on bone marrow may lead to Produce small erythrocyte with low hemoglobin concentration as the results of Hb and PCV showed in L-thyroxin treatment compared with control, Where hyperthyroidism interferes with iron metabolism the influence of thyroid hormones on the synthesis/ regulation of hepcidin an important regulator of iron metabolism ²¹, Increased red blood cells may be behind a decrease in ESR in this group. Also this great production of red blood cells in bone marrow may have significantly reduced the production of WBC in this treatment as our results showed. This corresponds to a study of ²², who concluded that treatment of rats with L-thyroxin for²⁸ days decrease WBC count, Because of hyperthyroidism cause depressed in total leucocytes with relative decrease in neutrophil³.

 

In addition thyroid disorders are usually accompanied with malnutrition and an unhealthy decreases in body weight^23,24, this case is most severe in hypothyroidism who developed by carbimazole drug may be because its’ bad taste and its effect on the digestive system leads to malnutrition^25,26, in T1 treatment observed decrease in food and water consumption, There for the decrease in ESR in T1and T2 may be due to some chemical and physical changes in the properties of plasma in some cases, Like malnutrition and oxidative stress caused by thyroid disorders^4,5 that mean changes in nature, quantity, density and viscosity of plasma, in particular increased plasma density due to drought or oxidative stress all this leads to increased density of plasma, which in turn causes lower because of the difficulty of deposition of blood cells into the test tube.

 

Many studies confirmed that thyroid disorders cause damage to the body organs^27,28. By inflammation and oxidative stress for example carbimazole induced Cholestatic Hepatitis and serositis²⁹, and the Hepatitis is associated with elevated WBC³⁰, that maybe explained the significant increase of WBC in carbimazole treatment specially in neutrophils as the first line of defense against the body against inflammations.

 

Conversely T4 and T5 treatments in which administrated GSE Synchronized with both drugs giving showed significant improvement in RBC, PCV, HB, ESR WBC, NE, LY compared with T1 and T2, GSE can contribute to this improvement first by antioxidant ability of grape seeds extract through some of its compounds like Phenolic, Flavonoids and Non-flavonoids which are known for their high ability to control free radicals ^31-33, this protect cells and tissues from the risk of organs damage like kidney protection caused by free radicals which occurs due to thyroid disorders and its drugs^34,35, And thus can perform its functions efficiently like erythropoietin production, As Flavonoids play an important roles in the defense against biotic and abiotic stresses³⁶.

 

Also GSE rich with Procyanidins which performed very well as anti-inflammatory, that maybe explained WBC count decrease in T4 (Carbimazole + GSE treatment).

 

The second way is grape extract contains proteins, amino acids, Glycosides, vitamins, Lipids, Carbohydrates, Organic acids, Phenolic acids and minerals ^37,38, this makes GSE has high nutritional value, thus addressing malnutrition caused by thyroid disorders, it Supports the cells with necessary materials to make the proteins and building tissues, thus contributing to the formation of blood elements and improving the plasma state.

 

ACKNOWLEDGEMENT:

The author is grateful to the Deanship of education and Veterinary and the Department of biology / Al-Qadisyiah University to provide all facilities for the success of this work

 

CONFLICT OF INTEREST:

The authors declare no conflict of interest.

 

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Received on 04.05.2019           Modified on 11.06.2019

Accepted on 01.07.2019         © RJPT All right reserved