بررسي اثر محفاظتي ويتامين C بر روي اسپرماتوسيت ها در موش هاي صحرايي نر مبتلا به صرع شده با پنتلين تترازول به روش ايمونوهيستوشيمي

Evaluation of the Protective Effect of Vitamin C on Spermatocytes in Rats with Pentylenetetrazole-Induced Epilepsy Using Immunohistochemistry

ﻣﻘﺪﻣﻪ: از آن ﺟﺎﯾﯽ ﮐﻪ در ﻣﻮرد ﺗﺄﺛﯿﺮ وﯾﺘﺎﻣﯿﻦ C ﺑﺮ روي اﺳﭙﺮﻣﺎﺗﻮﺳﯿﺖ ﻫﺎي ﻣﻮش ﻫﺎي ﻣﺒﺘﻼ ﺑﻪ ﺻﺮع، ﮔﺰارﺷﯽ ﻣﺸﺎﻫﺪه ﻧﺸﺪ؛ اﯾﻦ ﻣﻄﺎﻟﻌﻪ، ﺑﺎ ﻫﺪف ارزﯾﺎﺑﯽ ﻧﻘﺶ ﻣﺤﺎﻓﻈﺘﯽ وﯾﺘﺎﻣﯿﻦ C ﺑﺮ روي ﺗﻌﺪاد و ﺳﺎﺧﺘﺎر اﺳﭙﺮﻣﺎﺗﻮﺳﯿﺖ ﻣﻮش ﻫﺎي ﻣﺒﺘﻼ ﺑﻪ ﺻﺮع ﺷﺪه ﺑﺎ ﭘﻨﺘﻠﯿﻦ ﺗﺘﺮازول اﻧﺠﺎم ﺷﺪ. روش ﻫﺎ: در اﯾﻦ ﻣﻄﺎﻟﻌﻪ، 40 ﺳﺮ ﻣﻮش ﻧﺮ ﺑﻪ ﺻﻮرت ﺗﺼﺎدﻓﯽ ﺑﻪ 5 ﮔﺮوه 8ﺗﺎﯾﯽ ﺗﻘﺴﯿﻢ ﺷﺪﻧﺪ. ﮔﺮوه ﺷﻢ )اول( ﻣﺒﺘﻼ ﺑﻪ ﺻﺮع ﺷﺪه ﺑﺎ ﭘﻨﺘﻠﯿﻦ ﺗﺘﺮازول )40 ﻣﯿﻠﯽ ﮔﺮم/ﮐﯿﻠﻮﮔﺮم ﺳﺮم ﻓﯿﺰﯾﻮﻟﻮژي(، ﮔﺮوه ﻫﺎي دوم، ﺳﻮم و ﭼﻬﺎرم ﺗﺠﺮﺑﯽ ﻣﺒﺘﻼ ﺑﻪ ﺻﺮع ﺷﺪه ﮐﻪ دزﻫﺎي 125، 25 و 500 ﻣﯿﻠﯽ ﮔﺮم/ﮐﯿﻠﻮﮔﺮم وﯾﺘﺎﻣﯿﻦ C درﯾﺎﻓﺖ ﮐﺮدﻧﺪ. ﮔﺮوه ﭘﻨﺠﻢ، ﮔﺮوه ﺷﺎﻫﺪ ﻣﻨﻔﯽ ﺑﻮد ﮐﻪ ﻓﻘﻂ ﺳﺮم ﻓﯿﺰﯾﻮﻟﻮژي داﺧﻞ ﺻﻔﺎﻗﯽ درﯾﺎﻓﺖ ﮐﺮدﻧﺪ. ﭘﺲ از دوره ي درﻣﺎن، ﺑﯿﻀﻪ ي راﺳﺖ ﺣﯿﻮاﻧﺎت، ﺑﺎ ﺑﯿﻬﻮﺷﯽ ﻋﻤﯿﻖ ﺧﺎرج و ﭘﺲ از ﭘﺎﺳﺎژ ﺑﺎﻓﺘﯽ و رﻧﮓ آﻣﯿﺰي ﻫﻤﺎﺗﻮﮐﺴﯿﻠﯿﻦ- اﺋﻮزﯾﻦ، ﺷﻤﺎرش اﺳﭙﺮﻣﺎﺗﻮﺳﯿﺖ اﻧﺠﺎم ﮔﺮﻓﺖ. ﺗﻐﯿﯿﺮات رﯾﺨﺖ ﺷﻨﺎﺳﯽ ﻧﻤﻮﻧﻪ ﻫﺎ ﺑﺎ اﺳﺘﻔﺎده از روش ﻫﯿﺴﺘﻮﻟﻮژي و اﯾﻤﻮﻧﻮﻫﯿﺴﺘﻮﺷﯿﻤﯽ اﻧﺠﺎم ﺷﺪ. ﺑﺮاي ﺑﺮرﺳﯽ ﺗﻐﯿﯿﺮات ﻣﻌﻨﯽ دار ﺑﯿﻦ ﻧﻤﻮﻧﻪ ﻫﺎي ﻣﻮرد و ﺷﺎﻫﺪ، داده ﻫﺎ ﺑﺎ آزﻣﻮن ANOVA واﮐﺎوي ﺷﺪ. ﯾﺎﻓﺘﻪ ﻫﺎ: ﻣﯿﺎﻧﮕﯿﻦ ﺗﻌﺪاد اﺳﭙﺮﻣﺎﺗﻮﺳﯿﺖ ﻣﻮش ﻫﺎي ﮔﺮوه ﻣﺒﺘﻼ ﺑﻪ ﺻﺮع ﺷﺪه در ﻣﻘﺎﯾﺴﻪ ﺑﺎ ﮔﺮوه ﺷﺎﻫﺪ ﮐﺎﻫﺶ ﯾﺎﻓﺖ )0/001 < P(. اﻓﺰاﯾﺶ ﻣﻌﻨﯽ داري در ﻣﯿﺎﻧﮕﯿﻦ ﺗﻌﺪاد ﺳﻠﻮل ﻫﺎي اﺳﭙﺮﻣﺎﺗﻮﺳﯿﺖ ﻣﻮش ﻫﺎي ﻣﺒﺘﻼ ﺑﻪ ﺻﺮع ﺷﺪه ﮐﻪ 250 ﻣﯿﻠﯽ ﮔﺮم/ﮐﯿﻠﻮﮔﺮم وﯾﺘﺎﻣﯿﻦ C ﻣﯽ ﮔﺮﻓﺘﻨﺪ، در ﻣﻘﺎﯾﺴﻪ ﺑﺎ ﮔﺮوﻫﯽ ﮐﻪ 125 ﻣﯿﻠﯽ ﮔﺮم/ﮐﯿﻠﻮﮔﺮم وﯾﺘﺎﻣﯿﻦ C و ﯾﺎ ﺳﺮم ﻓﯿﺰﯾﻮﻟﻮژي درﯾﺎﻓﺖ ﻣﯽ ﮐﺮدﻧﺪ، دﯾﺪه ﺷﺪ )0/010 < P(. ﺗﻐﯿﯿﺮات رﯾﺨﺖ ﺷﻨﺎﺳﯽ ﺑﺮ روي اﺳﭙﺮﻣﺎﺗﻮﺳﯿﺖ ﻫﻤﭽﻮن ﻣﺘﺮاﮐﻢ ﺷﺪن ﻫﺴﺘﻪ و اﺳﯿﺪوﻓﯿﻠﯽ در ﺳﯿﺘﻮﭘﻼﺳﻢ ﺳﻠﻮل ﻫﺎي اﺳﭙﺮﻣﺎﺗﻮﺳﯿﺖ در ﻣﻮش ﻫﺎي ﻣﺒﺘﻼ ﺑﻪ ﺻﺮع ﺷﺪه، ﻧﺴﺒﺖ ﺑﻪ ﮔﺮوه ﺷﺎﻫﺪ ﺑﯿﺸﺘﺮ ﻣﺸﺎﻫﺪه ﺷﺪ. ﻧﺘﯿﺠﻪ ﮔﯿﺮي: ﻧﺘﺎﯾﺞ ﻣﻄﺎﻟﻌﻪ ي ﺣﺎﺿﺮ ﻧﺸﺎن داد ﮐﻪ اﻓﺰودن وﯾﺘﺎﻣﯿﻦ C، اﺛﺮات ﺳﻤﯽ ﭘﻨﺘﻠﯿﻦ ﺗﺘﺮازول را ﺑﺮ روي اﺳﭙﺮﻣﺎﺗﻮﺳﯿﺖ ﻫﺎي ﻣﻮش ﻫﺎي ﻣﺒﺘﻼ ﺑﻪ ﺻﺮع ﮐﺎﻫﺶ ﻣﯽ دﻫﺪ.

Background: There has not been yet reports regarding the protective effect of vitamin C on spermatocytes in epileptic rats. The present study aimed to evaluate the protective role of vitamin C on numbers and structure spermatocytes in rats with pentylentetrazol-induced epilepsy. Methods: Forty male rats were randomly divided into five groups of 8, and were kindled with pentylentetrazol (40 mg/kg). The first group (sham) received normal saline. The second, third, and forth experimental groups treated with 125, 250, and 500 mg/kg of vitamin C, respectively. The negative control group was given only normal saline via intraperitoneal injection. All rats were then deep anesthetized and sacrificed, and their right testis were dissected. Histological sections were prepared, and stained with hematoxylin-eosin (H/E). The number of spermatocytes in sections were counted, and morphological changes were studied using histology and immunohistochemical methods. Data were analyzes using ANOVA test to determine significant differences between cases and controls. Findings: There was a significant decrease in spermatocytes numbers in epileptic rats compared with intact control group (P < 0.001). The spermatocytes significantly increased in rats, which received 250 mg/kg vitamin C compared to those received 125 mg/kg vitamin C or normal saline (P < 0.010). The morphological changes such as dense nuclei and more acidophilic cytoplasm were observed in epileptic rats compared to control groups. Conclusion: The present study showed that vitamin C supplementation would protect spermatocytes by decreasing the neurotoxin effects of pentylentetrazol in epileptic rats.