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The effect of Thymus vulgaris on immune response and growth performance of albino rats.

INTRODUCTION

Growth means the relative irreversible time changes in the measured dimensions. The dimensions includes the size (length or height) and total body weight. True growth should include increase in weights of different tissues like the muscles, bone as well as internal organs, excess fat deposited in the fat depot in the body occurs in cases of feeding on imbalanced food containing excess carbohydrate, which will be stored in the body as fat [1].

Buttler and Leroith [2] added that there are many factors contribute for normal growth: genetic affect the growth through intrinsic regulators and extrinsic factors, also the plane of nutrition which provide the living organism with the building materials essential for growth, also the protein is an essential factor in the growth of young after birth as example milk supplies the young individual with protein that stimulate growth, also rabbits grow faster than chickens because it depend on milk in early life, sex, also affect growth in two folds, direct due to gene and indirect due to sex hormones, also vitamins and minerals act as cofactors required to utilize the food for growth.

Growth hormone has been believed to be secreted primarily during the period of growth, but then disappeared from the blood at adolescence, this has proved to be untrue. After adolescence, secretion decreases slowly with aging, finally falling to about 25 percent of the adolescent level in very old age [3]. They added that several factors related to the person's state of nutrition or stress are known to stimulate secretion: starvation, hypoglycemia exercise, excitement and trauma, also it has been found that growth hormone causes the liver and other tissues as muscles to form several small proteins called somatomedins, many of the somatomedin effect on growth are similar to the effects of insulin on growth, they are called insulin like growth factor.

Shaban [4], reported some pharmaco-dynamic studies of Thymus vulgaris plant, that Thymus vulgaris is used in folk medicine since ancient times and still used in modern era, and it contain many products as alkaloids, carbohydrates and glycosides, resins, saponins, tannins and unsaturated sterols and triterpenes and treatment of rats with extract of Thymus vulgaris at dose of 250 and 500 mg/kg b.w. protects the gastric mucosa against damaging effect and the prolonged administration of Thymus vulgaris for 2 months improved the body weight.

Osman, [5] reported the effect of thyme and garlic essential oils and their combination on the performance of Japanese Quail, growth digestibility, carcass traits hematological indices, they conclusion of the research indicated that Thyme oil and garlic had showed improvement on growing Japanese Quail performance, increased the antioxidant level and promote immune health. Shetti, [6] reported that adding thyme to wheat flour enhanced the natural antiodixant and nutritive value of treated bread. The treated bread was healthy which lowered the blood lipids and cholesterol. Abdel Fatah, [7] stated that Thymus induced antimicrobial activity and posses anti pain action [8], also due to its content of selected vitamins and trace elements which support immune function [9].

Reactive oxygen species (ROS) are capable of reversibly or irreversibly damaging many of the biological molecules including phospholibids, protein and DNA that have impact on cell activities as well as membrane functions and structure and DNA mutations [10]. The generation of ROS has been implicated in the pathogenesis of many diseases include cancer and inflammatory disorder [11]. Halliwell, [12] showed that shift to a more oxidative state might result in uncontrolled lipid peroxidation, protein oxidation and ultimately cell death. For these reasons, the search for novel substances that defense against ROS must be the target of many studies including this study. Several classes of compounds, such as protein, peptide, lipopolysaccharide, glycoproteins, lipid derivatives have been classified as molecules that have potent effects on the immune system [13].

Aim of the study:

This study was aimed at describing immunological and growth changes in response to Thymus administration to albino rats.

Hypothesis:

It was hypothesized that in response to Thymus administration to albino rats, they would show sign of immuno elevation and growth performance.

MATERIALS AND METHODS

The experimental animals used throughout the work were healthy male adult albino rats Sprague-Dawley strain weighing 190 [+ or -] 10 g, it was supplied by the breading unit of the Egyptian organization for Biological products and vaccines (Helwan, Egypt).

Animal trial:

Total number of healthy thirty albino rats were started for experimentation in animal biological room related to physiology department, Fac. of Veterinary Medicine, Zagazig Univ., where all rats were randomly housed individually in stainless steel cages with constant controlled environments: Temperature 25[degrees]C [+ or -] 5[degrees]C, and 12/12 hrs light/dark cycle were held, all rats were offered balanced diet with drinking water adlibitum. Before the experimental period all rats were held for 7 days as adaptation period and throughout the experiment, the rats were divided into two groups each was composed of 15 rats, the control and experimental groups, the experimental group administered a single oral daily dose thyme extract for two months at a dose level of 250 mg/kg bw [4].

Tools and devices used:

--Syringes.

--Test tubes with covers.

--Digital balance weight (satorus).

--Coleman, ices, Deep freezer.

--Kits for SOD, MDA, growth hormone.

--ELISA technique and spectrophotometer.

--Radial plates for immunoglobulins (IgG, IgA, IgM).

Measuring loop for reading:

The recommended dose for Thymus vulgaris 250 mg/kg b.w. daily for two months. Thymus vulgaris contain minerals and vitamins, vitamins B Complex, Vit A, Vit C, Vit K.

Minerals:

Potassium, Iron, Magnesium, Calcium, Manganese, Selenium.

Also volatile oils like thymol. Antioxidants, flavenoids including leuteoline, Nariginine, Thioumounin.

Statistical Analysis:

The data were analyzed using the statistical package for social science (SPSS) were characterized by their mean and standard deviations. Student "t" test was used for statistical analysis of the results and values at 0.05 level is considered statistically significant [14].

Results:

Table (1): Basic SOD, Malondialdehyde, immunoglobulins in control and experimental groups of albino rats indicated non significant changes of the parameters.

Table (2): Basic growth hormone, body weight in control and experimental albino rats revealed non significant changes of the parameters.

Table (3): The effect of thyme administration on SOD, MDA, Immunoglobulins revealed a significant increase of SOD, immunoglobulins and decreased MDA concentrations.

Table (4): The effect of Thyme administration on growth hormone, body weight of albino rats revealed a significant increase in both variables.

Discussion:

Superoxide anions are produced at any location where an electron transport chain in present, and hence O2 activation may occur in different compartments of the cell [15]. Therefore it is not surprising to find SOD's in all these subcellular locations including the plant roots which are used to scavenge these radical species [16].

Table (3) indicated that Thymus administration increased SOD activity and suppress Malondialdehyde, thus suppress the activity of the free radicals ([O.sub.2.sup.-], OH and [H.sub.2][O.sub.2]) produced. This was in accordance to Habib et al. [10]. Mohamed et al. [17], Packer [18]. In the present study, Thymus showed SOD activity, this enzyme uses [O.sub.2] as a substrate converting it into H2O2, which can be used by other antioxidants like catalase and glutathione peroxidase. This indicated the protective effect of the Thymus. Table (3) revealed an increased antibodies product IgG, IgA, IgM in response to Thymus administration. This result confirm the ability of the Thymus administration to stimulate B-cells to produce specific antibodies. Since the specific immunity to cells includes generation of antibodies, promotion of inflammatory responses leading to kill the pathogens [19]. Alscher et al. [16] reported the role of superoxide dismutase in controlling oxidative stress in plants, and in related areas in molecular biology [20]. Also Al Zahraa et al. [21] stated that Thymus may improve natural antioxidant activity and the treated bread with Thymus lower lipids and cholesterol and improve health, The data of this research was in accordance with those of Alshaimaa [22], Hassan [23], Hasnaa [24], Trinchieri [25] and Udintsev et al. [26]

Data presented in table (3) indicated an increased body weight of albino rat after Thymus administration together with an elevation of growth hormone registered. This result is in accordance with that of Osman, (5); Hasnaa, [24]; Hassan, [27].

Guyton and Hill, [3] stated that all the major anterior pituitary hormones, except for growth hormone, exert their principal effects by stimulating target glands. Growth hormone, in contrast to other hormones, does not function through a target gland but exerts its effects directly on all or almost all tissues of the body. They added that in comparison of a rat gain injected daily with growth hormone with that of a normal littermate. In the early stages of development, all organs of the treated rats increase in size, after adulthood was reached, most of the bones stopped lengthening, but many of the soft tissues continue to grow.

Cunningham [28] and Ganong [29] stated that growth hormone perform its effects on target tissues through liberation of somatomedin C also called [IGF.sub.1] and [IGF.sub.2]. These polypeptides are secreted from liver and trivial amounts from other tissues like muscles. He added that vitamins and minerals like vit. [B.sub.1], complex, [B.sub.12], vit. E, C, A, together with calcium, phosphorus selenium, magnesium and manganese, iron, copper, cobalt, zinc, molybdenum all these minerals and trace are very essential to growth, which are constituents of the Thymus vulgaris which denote the important role by this plant in promoting growth of the albino rats of this study.

The increased growth remarked after Thymus administration may be due to the antioxidant action of Thymus together to its content of vitamins and minerals all of which help in increasing growth and body weight reported in this study, which indicate that Thymus administration increased growth hormone and growth hormone in turn enhances almost all facets of amino acid uptake and protein synthesis by cells, while at the same time reducing the breakdown of protein inducing increase in body weight [30; 31, 32; 33; 34].

In conclusion, the results showed that Thymus had showed encouraging improvement on growing albino rats performance. Using Thymus as feed additive increased the antioxidant level which can help to control oxidative stress and imbalances and promote better immune health. This means that Thymus would be safe for the rats to consume, which in turn helping the lab animal to exhibit better health and growth, due to Thymus content of different vitamins, minerals and oil like thymol, antioxidants, flafanoid compound. From the previous discussion. The hypophesis, that in response to Thymus administration to albino rats, they would show sign of immuno elevation and growth performance. The hypophesis has been realized.

REFERENCES

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[2] Buttler, A and D. Leroith, 2001. Control of growth by the somatotropic orig. Ann Rev Physiol., 63: 141.

[3] Guyton, A and J. Hall, 2006. Textbook of medical physiology. El sevier, Saunders, USA.

[4] Mohamed Shaban, 2016. Some pharmacodynamic studies of Thymus vulgaris plant. PhD, Fac. of Vet. Med., Beni Suef Univ.

[5] Abo Mahara Osman, 2015. Effect of thyme and garlic essential oils and their combination on the performance of Japanese Quails. PHD, Fac. of Agriculture, Alex Univ.

[6] Al-Mamoori Shetti, 2015. Changes of antioxidants during making of bread. PHD, Fac. of Agriculture, Mansoura Univ.

[7] Ahmed Abdel Fatah, 2015. Studies on antimicrobial activity of some plant extract against staph and candida with mastitis. Fac. of Oral, Dental Medicine, Cairo Univ.

[8] Hesham Abdullah, 2015. Clinical, Radiographic and hitological effect of propolis, Nigella, Thymus formocresol for pulpotomy of vital molars. PhD Fac. of Oral, Dental Medicine, Cairo Univ.

[9] Majeny, H., 2007. Selected vitamins and trace elements support immune function by strengthening epithelial barriers and hormonal immune response. British J Nutr. 9: 24.

[10] Salem Habib, T. Elshahat, A. Mohamed, 2005. Antihemolytic and antitumor effects of Rhaphnus Sativa with M-m SOD like activity. EJBMB, pp: 272.

[11] Sokol, R and E. Hoffnberg, 1996. Antioxidant in pediatric gastro-intestinal disease. Pediat Gastroenterol, 13: 471.

[12] Halliwell, B., 1999. Antioxidant defense mechanisms: Free Radio. Biol Med., 31: 261.

[13] Ooi, V and F. Liu, 2000. Immunomodulation and anticancer activity of polysaccharide-protein complexes. Curr. Med. Chem., 7: 715.

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[15] Elstner, E., 1991. Mechanism of oxygen activation in different compartment of plant cells. Am Soc of plant physiologists, p: 13.

[16] Alscher, R., N. Erturk and L. Heath, 2002. Role of superoxide dismutase in controlling oxidative stress in plants. J. Exp. Botany, 53: 1331.

[17] Mohamed, K., Z. Reham, A. Nora, 2014. Anti ulcer and gastroprotective effects of fenugreek, ginger, peppermint oils in experimentally induced gastric ulcer in rats. J of Chemical and Pharm. Res., p: 468.

[18] Packer, L., 1993. Principles and applications in: Organic photochemistry. Academic Press, London, p: 324.

[19] Han, S., S. Park, K. Lee, 2001. Polysaccharide isolated from the radix of platycodon grandiflorum selectively activated B cells and macrophages but not T cells. Int. Immunopharmacology, 1: 1969.

[20] Fridovich, I.C., 1986. Superoxide dismutases. Molecular Biology, 58: 61.

[21] Fatma Alzahraa, 2015. Pharmacognostical study and biological investigation of certain Thymus species. PhD, Cairo Univ., Fac. of Pharmacy.

[22] Alshaimaa, A., 2013. Biochemical effect of pirimiphos methyl and lipid peroxidation and antioxidant enzymes of rats and ameliorating role of certain antioxidants. Phd, Banha Univ. Fac. of Vet Medicine.

[23] Reem Hassan, 2008. Effect of some organic element on the growth of Thymus Plant. Master, Environmental Research Institute, Ain Shams Univ.

[24] Hasnaa, S., 2006. Biochemical studies and physiology of Thymus plant. Fac of Agriculture, Cairo Univ.

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[26] Udintsev, S., S. Krylova and T. Fomina, 1992. The enhancement of the efficacy of adriamycin by using hepatoprotection of plant origin in metastases of adenocarcenoma to the liver in mice. Vopr Onkol, 38: 1217.

[27] Mervat Hassan, 2015. Evaluation of the anti-inflammatory activity of some dietary supplement in chronic inflammation model. Fac. of Agriculture, Cairo Univ.

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[29] Ganong, W., 2000. Review of Medical Physiology. Lange Medical Book, Mcgraw Hill.

[30] Cummings, D and G. Merriam, 2003. Growth hormone therapy in adults. Annu Rev Med., 54: 513.

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[32] Goldspink, G., 2004. Age related muscle loss and progressive dysfunction in growth factor signaling. Ann NY Acad Sci., 1019: 294.

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Ryeaan Abdel Moneim Sayed

Household Economy Department (Nutrition), Faculty of Specific Education, Port Said University, Port Said, Egypt.

Address For Correspondence:

Ryeaan Abdel Moneim Sayed Household Economy Department (Nutrition), Faculty of Specific Education, Port Said University, Port Said, Egypt.

E-mail: Rayyan8@gimail.com

Received 12 May 2016; Accepted 28 June 2016; Available online 28 July 2016
Table 1: Basic SOD, Malondialdehyde, immunoglobulins concentrations
in control and experimental groups.

Group          SOD                 Malondialdehyde
               [micro]/ml          Mmol/ml

Control        121 [+ or -] 10.1   63.2 [+ or -] 1.7
Experimental   122 [+ or -] 11.3   64.1 [+ or -] 1.3

Group          Immunoglobulin

               IgG                 IgM

Control        0.20 [+ or -] 0.1   0.37 [+ or -] 0.2
Experimental   0.21 [+ or -] 0.2   0.35 [+ or -] 0.1

Group          Immunoglobulin

               IgA

Control        0.04 [+ or -] 0.01
Experimental   0.05 [+ or -] 0.1

Non significant level P < 0.05

Table 2: Basic growth hormone, body weight in control and
experimental groups.

Group          Growth hormone     Body weight/g
               Pg/ml

Control        3.5 [+ or -] 0.8   190 [+ or -] 18.1
Experimental   3.8 [+ or -] 0.9   1.91 [+ or -] 19.2

Non significant level P < 0.05

Table 3: Effect of administration of thyme on superoxide dismutase,
malondialdehyde and immunoglobulin concentrations.

Group         SOD                   Malondialdehyde
              m/ml                  Mmol/ml

Control       124.5 [+ or -] 11.3   6 5.8 [+ or -] 1.6
Thyme group   212.3 [+ or -] 22.1   41.3 [+ or -] 1.8

Group         Immunoglobulin

              IgG                 IgM

Control       0.19 [+ or -] 0.1   0.36 [+ or -] 0.2
Thyme group   0.41 [+ or -] 0.2   0.45 [+ or -] 0.03

Group         Immunoglobulin

              IgA

Control       0.04 [+ or -] 0.01
Thyme group   0.16 [+ or -] 0.1

Significant level P < 0.05

Table 4: Effect of administration of Thyme on growth hormone and
body weight.

Group          Growth hormone     Body weight/g
               Pg/ml

Control        3.6 [+ or -] 0.7   238 [+ or -] 17.3
Experimental   5.9 [+ or -] 0.9   250 [+ or -] 21.4

Significant level P < 0.05
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Author:Sayed, Ryeaan Abdel Moneim
Publication:Advances in Environmental Biology
Article Type:Report
Date:Jun 1, 2016
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