الفهرس 
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Abstract
Epilepsy is a widely spread disease with a recurrent spontaneous seizure. Several epidemiological studies indicate mortality in epileptics. Most of the increased risk is directly related to the cause of epilepsy itself. Sudden unexplained death in epilepsy patients is the most important cause of epilepsy-related deaths especially in the young and middle-aged groups which is connected with seizures, particularly tonic-clonic seizures. Anti-epileptic drugs (AEDs) are used to reduce the frequency of seizures. Valproic acid (VPA) is a drug of first choice and one of the most frequently prescribed antiepileptic drugs worldwide for the therapy of generalized and focal epilepsies, including special epileptic. This is done in high doses and for long time, which is always accompanied with side effects. This study aims to use Bee Venom (BV) as a natural product with anti-convulsant effect and comparing its effect with the used VPA to reduce the oxidation and reduce the drugs-side effects.
In this study thirty-two white adult male Sprague Dawley rats weighing about 150 -180 g were used. Rats were given access to water and a well-balanced standard meal daily. The rats were randomly classified into four groups eight rats/group as follows:
(1) Control group (C) received only standard diet, free access to sterile water and orally fed with (PBS, pH 7.4; 0.2 ml/rat) using intragastric intubation at intervals parallel to the treated groups.
(2) Positive epileptic group, epileptic control “EP group” was treated with 300 mg pilocarpine hydrochloride/Kg. b.wt of rats.
(3) Valproic acid-treated epileptic group (EP-VPA group): this group was injected with 300 mg/kg of pilocarpine hydrochloride injection as described previously, and then orally fed with 500 mg VPA® /Kg. b.wt of rats, dissolved in PBS (pH 7.4; 0.2 ml/rat). VPA® inoculation was performed twice/week for four consecutive weeks, using intragastric intubation at intervals parallel to other groups for four consecutive weeks.
(4) Bee venom-treated epileptic group (EP-BV group): It was injected with 300 mg/kg of pilocarpine hydrochloride injection as described previously, and then injected intradermal (i.d.) with 10 µg/animal once every 3 days for four consecutive weeks of pure BV dissolved in PBS (pH 7.4).
Behavioral tests were carried out and analyzed by two investigators unaware of the treatment of animals. After that, the brain was excised and dissected for biochemical and histopathological studies.
Moreover, liver and testis were obtained for further histological investigation. Epileptogenesis was assured by measuring the level of electrolytes and oxidative stress in both serum and hippocampus tissues. On the other hand, serum liver functions and sexual hormonal is measured. Hippocampus stained by Golgi, Hematoxylin and Eosin and glutaraldehyde staining. In addition, liver and testis tissues fixed and stained for histological light and electron investigation.
The present investigation obtained the following results:
1- The results of locomotors activity for Open field test (OFT), Hot plate test (HPT) and Forced swimming test (FST) in EP-BV showed good results when compared to other groups.
2- These imbalanced levels of electrolytes (Na+, K+, Ca2+ and CL-) were corrected after treatment with BV. There is an increase in intracellular potassium and chloride whereas sodium and calcium decreased.
3- Liver functions are routinely monitored. BV decreased plasma liver enzymes (ALT and AST) and increase the total proteins comparing with EP group.
4- BV showed a significant improvement in LH, FSH, testosterone levels.
5- The pro-oxidation mechanism represented by Lipid Peroxidation, Nitric Oxide and the antioxidant defense mechanism (Catalase, Superoxide Dismutase, Glutathione Peroxidase) showed ameliorative results in epileptic-treated groups when compared to the positive pilocarpine epileptic one.
6- Pilocarpine induced several histopathological alterations in hippocampus including degenerated and decrease of pyramidal cells number, deeply stained pyramidal cells with irregular pyknotic nuclei, while epileptic group with Depakine® revealed most pyramidal cells with large vesicular nuclei and a smaller number of pyramidal cells with darkly stained nucleus but BV showed marked amelioration of this histopathological changes that is assured by light microscope.
7- In Golgi–Cox-stain, pyramidal cells of epileptic group showed less dendritic branching compared with that of the control group. Epileptic group treated with VPA revealed moderate increase in pyramidal cells’ dendritic branching, while epileptic-treated group with BV show marked pyramidal cells’ dendritic branching similar to the control group.
8- An electron micrograph of the hippocampus nerve cell of epileptic animals showed degenerated pyramidal cell with irregular nuclear envelop nucleus, vacuolated cytoplasm, dilated RER, electron-dense mitochondria with ill-defined cristae and degenerated myelinated nerve fibres, while epileptic-treated with VPA and BV showed nearly normal pyramidal cell with nucleus, RER, ribosomes and mitochondria except oligodendrocyte with some vacuoles surrounded by myelinated nerve fibres.
9- Pilocarpine displayed several histological changes in testis as detachment of germ cells from the basement membrane, degenerated seminiferous tubules odema and marked decrease of sperms were observed. Treatment of epileptic animals with BV successfully induced great recovery and restoration of normal testicular configurations comparing with the epileptic rats and that assured by light and electron microscope.
10- Marked changes were revealed in epileptic-treated rats; these include congested blood vessels and sinusoids, vacuolated hepatocytes, fat cells with variable-sized, pyknotic nuclei and other karyolytic nuclei, binucleated cells, swollen hepatocytes, focal necrosis and mononuclear leukocytic infiltration. Other hepatocytes revealed marked deposition of collagen and electron-dense mitochondria appeared with ill-defined cristae. The liver sections of epileptic group treated with VPA demonstrated mild improvement except few vacuolated hepatocytes and slight hyperemic sinusoid while the liver sections of epileptic group treated with BV revealed an obvious amelioration of the hepatic cells with normal nucleus, mitochondria and glycogen. Also, Kupffer cell appeared with nearly normal nucleus. These results confirmed by using stained sections with H&E stain and an electron microscope.
In conclusion, BV can reduce the high rate of mental retardation and behavioral problems and exerting antioxidant effect. It attenuates neuronal inflammation in the hippocampus as well as severe histopathological and ultrastructure changes in the liver and testis. This is assured by its electrolyte balancing effect. BV can maintain liver functions and can ameliorate sexual performance by affecting sexual hormones. So, BV could confer beneficial effects against epileptogenesis due to its antioxidant effect.