"" Effect of Ginseng on the Testis of Subclinical Hypothyroidism Model in Adult Male Albino Rat

Research Article

Austin J Anat. 2017; 4(2): 1065.

Effect of Ginseng on the Testis of Subclinical Hypothyroidism Model in Adult Male Albino Rat

Issa NM and El-Sherif NM*

Department of Anatomy and Embryology, Faculty of Medicine, Menoufia University, Egypt

*Corresponding author: El-Sherif NM, Department of Anatomy and Embryology, Faculty of Medicine, Menoufia University, Egypt

Received: January 31, 2017; Accepted: February 13, 2017; Published: February 17, 2017

Abstract

Hyperstimulation of follicular cells using a goitrogen Propylthiouracil (PTU) caused Subclinical Hypothyroidism (SCH) that was associated with disturbances in adult male testicular functions. This work was carried out to study the probable relationship between follicular cells in drug-induced hypothyroidism caused by PTU and consequently its effect on testicular tissue and the possible modulating effect of ginseng. The present work was carried out on 40 adult male albino rats divided into four groups. Group I was the control group; Group II (Ginseng treated); Group III (SCH group) rats and Group IV was the treated group. Testes and thyroid glands were studied. Our results revealed the protective role of Ginseng on testis and thyroid tissue of experimentally induced hypothyroidism rat model.

Keywords: Subclinical hypothyroidism; Follicular cells; Testis

Introduction

Thyroid hormone plays an important role in testicular development and function. It has been established that Triiodothyronine (T3) regulates the maturation and growth of the testis, controlling Sertoli cells and Leydig cell proliferation and differentiation during testicular development in rats and other species [1]. Subclinical Hypothyroidism (SCH) could be considered as an elevated serum thyrotropic stimulating hormone (TSH) level associated with normal Thyroxine (T4) and Triiodothyronine (T3) levels with no clinical symptoms of hypothyroidism. This thyroid dysfunction occurs in 4-20% of the adult population, the possibility of SCH progression to clinical hypothyroidism is around 7% [2]. Hypothyroidism extremely affects reproductive functions including fertility, pregnancy and postnatal development in human and rat [3]. Rat SCH model was induced mostly by administration of antithyroid drug mostly Propylthiouracil (PTU) and Methimazole (MMI) [4]. PTU is a thiouracil-derived drug which can be used to treat hyperthyroidism through lowering the amount of thyroid hormone produced by the thyroid gland [5]. PTU inhibits conversion of T4 to T3 causing hypothyroidism [6]. Ginseng is considered as an aromatic herb that is commonly used in herbal medicine [7]. It contains saponins and high iodine content [8]. Ginseng saponins, which are known as ginsenosides, have been studied to be responsible for its medicinal effects, specifically anticancer, antihypertensive, antidiabetic, and antistress effects. Ginseng benefits all the endocrine system, and therefore the thyroid gland. As ginseng markedly increased epididymal sperm count, motility and hyperactivation in mice [9]. So, we studied its effect on the testis and thyroid of SCH model.

Methodology

Animals

Forty adult Westar male albino rats were used in this experiment, each weighting 150-200gm. Food and water were provided ad libitum and the rats were left for 7 days for acclimatization before use in the Anatomy Department, Faculty of medicine, Menoufia University. All aspects of animal care and treatment were carried out according to the local guidelines of the ethical committee for animal research.

Experimental plan

The animals were divided into four groups as the following:

Group I (Control): included ten rats were kept without any treatment throughout the experimental period.

Group II (Ginseng treated): included ten rats that received ginseng (Panax ginseng) a product of Pharco Pharmaceuticals, Alexandria, Egypt, was available in the form of capsules with the trade name ‘Ginseng 100’. Each capsule contained 100mg of the dried roots of Panax ginseng. The contents of the capsule was withdrawn by a syringe and dissolved in distilled water and given to the rats in a dose of 2mg/kg once daily for three weeks by gastric tube.

Group III (Subclinical hypothyrid SCH group): included ten rats that received oral propylthiouracil in a dose of 5mg/kg once daily for three weeks by gastric tube.

Group IV (Treated group): included ten rats with subclinical hypothyroidism that received oral treatment of ginseng in a dose of 2mg/kg dissolved in distilled water once daily for three weeks by gastric tube two weeks after the induction of subclinical hypothyroidism.

Hormonal assay

Two weeks after subclinical hypothyroidism induction, retroorbital blood samples of all rats were collected and plasma Thyrotropic Stimulating Hormone (TSH), Plasma Thyroxine (T4) and Plasma triiodothyronine (T3) were measured. Sera were separated and stored at −20°C until the hormonal assay was performed. Serum levels of T3 and T4 were determined by chemilumines-cence immunoassay using Amerlite kits (Bunkyo-ku, Tokyo, Japan) [10]. Serum levels of TSH were assayed by radioimmunoassay [11]. Serum levels were measured at the Clinical pathology Department, Faculty of Medicine, Menoufia University.

Epididymal spermatozoal study

Twenty-four hours after the last treatment, animals were euthanized under anesthesia, according to the time of the experiment. The epididymal content of each rat was obtained immediately by cutting the tail of the epididymis and squeezing it gently to obtain fresh undiluted semen in a clean Petri dish to proceed for epididymal spermatozoal examination.

Histological study

Thereafter, animals were killed by cervical dislocation under ether anesthesia. The testes and thyroid gland of each rat were immediately excised and prepared for histological study. The right testis and the right thyroid lobe was cut into small pieces (about 1×1mm3) and fixed in 3% phosphate-buffered glutaraldehyde and was further processed and prepared for transmission electron microscopic examination [12] using a Joel 100 CX Electron Microscope (Joel, Tokyo, Japan) at the Electron Microscopy Unit, Faculty of Science, Menoufia University. Left testis samples were fixed in bouin’s solution while left thyroid samples were fixed in 10% formol saline and processed to prepare 5μm-thick paraffin sections for:

Testicular sections were incubated with rabbit polyclonal anti- Androgen Receptors (anti-AR) antibody. Fremont, California, USA (Thermo Scientific) [13] while thyroid sections were incubated with rabbit monoclonal Anti-Thyroglobulin antibody. Abcam’s RabMAb technology, USA.

Quantitative assessment

The following parameters were measured:

Five non-overlapping fixed fields, each 2mm2 in area, were taken using a Leica DMLB2/11888111 microscope equipped with a Leica DFC450 camera using the Leica C PLAN 10×0.22 objectives.

Statistical analysis

Data from the hormonal assay and immunohistochemical study were tabulated and expressed as mean±SD. They were fed into the computer using statistical package for the social sciences (SPSS, version 20; IBM SPSS, Chicago, Illinois, USA) software package [14]. Statistical analysis was carried out using one-way analysis of variance and the post-hoc test (Scheffe) for pair wise comparison. A level of significance of p<0.05 was considered to be statistically significant.

Results

There was no significant difference between Group I (Control) and Group II (Ginseng treated) rats in all the outcomes; therefore, these two groups were pooled in one group (Control).

Hormonal assay

The biochemical profiles of the three groups are shown in (Table 1). SCH was confirmed by measuring plasma T3, T4 and TSH levels. The serum TSH concentration in the SCH group was significantly increased compared with that in the control group.