Review of Global Medicine and Healthcare Research

Volume 3 No. 2 (2012) Publisher: DRUNPP Managed by: IOMC Group ISSN: 1986-5872

REVIEW OF GLOBAL MEDICINE AND HEALTHCARE RESEARCH (RGMHR) ISSN: 1986-5872

EFFECT OF PALM VITAMIN E ON SPERM MORPHOLOGY OF STREPTOZOTOCIN INDUCED DIABETIC RATS Jamaludin M. *, Daren K., Siti Balkis B. 1

Biomedical Science Programme, Faculty of Health Science, Universiti Kebangsaan Malaysia, 50300 Kuala Lumpur, Malaysia

Corresponding author: Jamaludin Mohamed, PhD; Programme of Biomedical Sciences, Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, MALAYSIA. Tel. No. : +603-9289 7632 | Email : [email protected] ABSTRACT

Diabetes mellitus is a common endocrine metabolic disorder, which results in the impairment of uptake and storage of glucose and reduced glucose utilization for energy purposes. Free radicals are very reactive chemical species, can cause oxidative injury. The initial and most essential step of the infertility evaluation is semen analysis. The role of vitamin E as the body’s primary lipid-soluble antioxidant makes it a good candidate for investigation of effects against reactive oxygen species (ROS). The aim of the present study was to investigate the morphological changes of sperm in streptozotocin induced diabetic rats treated with palm vitamin E (PVE). A total of 40 (250-280g) Sprague Dawley rats were used and divided into four groups randomly. To induce diabetes mellitus STZ 45 mg/kg was injected intravenously. The diabetic rats were divided into two groups: supplemented with PVE (200 mg/kg/day) and non-supplemented with PVE. The non-diabetic rats were also divided into two groups: supplemented with PVE (200 mg/kg/day) and non-supplemented with PVE. Following of four weeks of force feeding treatment, rats were sacrificed and sperm samples were taken from the cauda. For qualitative analysis of sperm histology, cauda is excised in petri dish containing Hank’s buffer solution. Sperm obtained from the cauda is analysed for sperm viability using eosin-nigrosin staining. As for blood glucose, the mean level of blood glucose for normal rats are 172.01, normal + PVE is 144.96, diabetic rats are 510.41 and for diabetic + PVE is 536.91mmol/L in the period of four weeks. Based on the staining observation there is notable improvement in the sperm analysis such as viability and morphology. This study suggested that PVE may be effective in reducing oxidative stress in sperm of STZ induced diabetic rats. It could be concluded from the present results that administration of PVE offers significant protection to diabetes-induced oxidative stress by decreasing lipid peroxidation and activating antioxidant enzymes in the testes, thereby ameliorating diabetes-induced suppressed reproduction in male rats. Keywords: Palm vitamin E, sperm, lipid peroxidation, diabetes

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REVIEW OF GLOBAL MEDICINE AND HEALTHCARE RESEARCH (RGMHR) ISSN: 1986-5872

1. Introduction Infertility is defined as a condition where a couple is unable to conceive after one year of unprotected intercourse and male infertility contributes largely to infertile couples (Jonathan et al. 2002). Majority of male infertility causes are due to deficient sperm production of unknown origin, environmental and nutritional factors (Sinclair 2000). Patients suffering from diabetes mellitus (DM) often develop reproductive disturbances including impotence and hypospermia (Kolodny, 1974). Diabetes mellitus is a common endocrine metabolic disorder, which results in the impairment of uptake and storage of glucose and reduced glucose utilization for energy purposes (Wild et al. 2004). Oxidative stress occurs in diabetes when products called free radicals and reactive oxygen species, which are formed in normal physiology but become deleterious when not quenched by antioxidants systems (Hayden & Tyagi, 2002; Fang et al. 2002; Dröge, 2001). Free radicals are very reactive chemical species, can cause oxidative injury to the living beings by attacking the macromolecules like lipids, carbohydrates, proteins and nucleic acids. Under normal condition, there is an established equilibrium between production of reactive oxygen species (ROS) and antioxidant system deactivates them (Halliwell, 2004). The initial and most essential step of the infertility evaluation is semen analysis. The evaluation of male fertility potential is based on the assessment standard semen variables which include sperm concentration, progressive sperm motility and sperm morphology. Of these semen variables, sperm morphology has been regarded as one of the most important for determining the quality of a semen sample and a male's fertility potential (MacLeod and Gold, 1951; Hartman et al., 1964; Eliasson, 1971; Menkveld and Kruger, 1996; Coetzee et al., 1998). The role of vitamin E as the body’s primary lipid-soluble antioxidant (Wang and Quinn, 1999) makes it a good candidate for investigation of effects against diseases of aging, especially those that involve ROS as a main component (Robert and John 2010). Alpha tocopherol the naturally occurring antioxidant in biological systems and is present in the cell membrane of various tissues including the intestines and stomach. Vitamin E prevents free radical-induced injury by blocking the free radical chain reaction (Kamsiah et al, 2006). The antioxidant function of α-tocopherol is well documented. But it is now evident that depending on the dose levels, α-tocopherol could exhibit pro-oxidant activity. More recently evidence has accumulated to show that vitamin E has biological functions other than its classical antioxidant activity (Traber & Packer, 1995; Azzi & Stocker, 2000).

2. Materials and methods 2.1 Chemicals Gluteraldehyde, Normal saline, Eosin Y 1%, Nigrosin 10% 2.2 Experimental Design

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REVIEW OF GLOBAL MEDICINE AND HEALTHCARE RESEARCH (RGMHR) ISSN: 1986-5872

Healthy male Sprague-Dawley rats were purchased from the Laboratory Animal Resource Unit, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur and kept one week for acclimation. The rats were maintained at a controlled temperature (25± 2) and natural light-dark cycle. After acclimatization, rats weighing 250 and 290g were divided into 2 groups and each group has 10 rats. The rats are housed in plastic cages (two rats per cage) with free access to food and water for 30 days. The rats is group I served as control and was given oral administration of Olein (10mg/kg/bw) and group II is induced with diabetes and given oral administration olein same as the control group Experimental diabetes was induced in 12 h fasted rats by single intravenous injection of streptozotocin (STZ) (45mg/kg body weight). Diabetes was confirmed in the overnight-fasted rats by measuring blood glucose concentration 72 h after injection with STZ. The rats with blood glucose above 240 mg/dl were considered to be diabetic and selected for the experiment (Palsamy et al. 2010). At the end of experience, all the animals were fasted overnight, weighed and sacrificed by cardiac puncture after a light anaesthesia with diethyl ether. 2.3 Body and organ weights Body weights of animals were recorded at the beginning and at the end of the treatment period. The animals were dissected and organs of the reproductive tract such as testes, seminal vesicles, ventral prostate, epididymides (caput and corpus) were excised, cleared of adhering fat and weighed. 2.4 Sperm Analysis Sperm viability was accessed by eosin-nigrosin staining. 10µl of sperm is placed on a glass slide followed by 2 drops of eosin Y and allowed for 30 seconds then 3 drops of nigrosin is placed and after 30 seconds a smear is made with a cover slip and allowed to air dry before viewing under light microscope.

3. Results

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REVIEW OF GLOBAL MEDICINE AND HEALTHCARE RESEARCH (RGMHR)

glucose oxidase (mmol\L)

ISSN: 1986-5872

a,b 29.77

40 35

a,c 28.30

30 25 10.457

8.184

Normal Group

N+PVE

20 15 10 5 0

Diabetic Group

D+PVE

Experimental Groups

Figure 1: The effect of treatment with PVE on blood glucose in the Experimental groups of rats a- Significantly different from the Normal group (negative control) P