ﺑﺮرﺳﯽ اﺛﺮ آﻟﮑﺎﻟﻮﺋﯿﺪ ﻫﺎرﻣﯿﻦ ﺑﺮ روي ﺑﻠﻮغ آزﻣﺎﯾﺸﮕﺎﻫﯽ اووﺳﯿﺖ ﮔﺎو

Investigation of the Effects of Harmine Alkaloid on Bovine Oocyte in Vitro Maturation

ﺑﻠﻮغ اووﺳﯿﺖﻫﺎ در ﺷﺮاﯾﻂ آزﻣﺎﯾﺸﮕﺎﻫﯽ ﺗﮑﻨﯿﮑﯽ اﺳﺖ ﮐﻪ ﻣﯽﺗﻮاﻧﺪ ﺑﺎﻋﺚ ﮐﺎﻫﺶ ﻫﺰﯾﻨﻪﻫﺎ و از ﺑﯿﻦ ﺑﺮدن اﺛﺮات ﺟﺎﻧﺒﯽ اﺳﺘﻔﺎده از ﮔﻨﺎدوﺗﺮوﭘﯿﻦﻫﺎ ﺑﺮاي ﻟﻘﺎح آزﻣﺎﯾﺸﮕﺎﻫﯽ ﺷﻮد. ﺗﺤﻘﯿﻖ و ﭘﮋوﻫﺶ در زﻣﯿﻨﻪ ﺗﻮﺳﻌﻪ و ﺑﻬﺒﻮد ﺷﺮاﯾﻂ ﮐﺸﺖ آزﻣﺎﯾﺸﮕﺎﻫﯽ ﺑﺮاي ﺑﻠﻮغ اووﺳﯿﺖ اﻧﺴﺎﻧﯽ ﺑﺴﯿﺎر ﺳﺨﺖ اﺳﺖ. اﻣﺮوزه اﺳﺘﻔﺎده از ﻣﺪل ﺣﯿﻮاﻧﺎت اﻫﻠﯽ ﭘﯿﺸﺮﻓﺘﻪﺗﺮﯾﻦ و ﻋﺎﻟﯽﺗﺮﯾﻦ ﺳﯿﺴﺘﻢ را ﺑﺮاي ﻣﻄﺎﻟﻌﻪ در زﻣﯿﻨﻪ ﺑﻠﻮغ ﻓﻮﻟﯿﮑﻮلﻫﺎي ﻧﺎرس در آزﻣﺎﯾﺸﮕﺎه ﻓﺮاﻫﻢ ﮐﺮده اﺳﺖ. ﺑﺎ ﺗﻮﺟﻪ ﺑﻪ ﻣﺎده ﻣﻮﺛﺮه و دﯾﮕﺮﺗﺮﮐﯿﺒﺎت ﻣﻮﺟﻮد در آﻟﮑﺎﻟﻮﺋﯿﺪ ﻫﺎرﻣﯿﻦ ﮐﻪ داراي اﺛﺮات ﻓﺎرﻣﺎﮐﻮﻟﻮژﯾﮏ ﻣﺘﻌﺪد از ﺟﻤﻠﻪ ﺿﺪ رادﯾﮑﺎل آزاد، ﺿﺪاﻟﺘﻬﺎب و اﺛﺮ ﺑﺮ ﺳﯿﺴﺘﻢ اﯾﻤﻨﯽ و ﻏﯿﺮه ﻣﯽﺑﺎﺷﺪ، ﺑﻨﺎﺑﺮاﯾﻦ در اﯾﻦ ﻣﻄﺎﻟﻌﻪ اﺛﺮات آﻟﮑﺎﻟﻮﺋﯿﺪ ﻫﺎرﻣﯿﻦ ﺑﺮ ﻣﯿﺰان ﺑﻠﻮغ ﺗﺨﻤﮏﻫﺎي ﮔﺎوي ﻣﻮرد ﺑﺮرﺳﯽ ﻗﺮار ﻣﯽ ﮔﯿﺮد. در اﯾﻦ ﻣﻄﺎﻟﻌﻪ ﺗﺨﻤﮏ ﻫﺎي آﺳﭙﯿﺮه ﺷﺪه ﺣﺪاﻗﻞ داراي ﺳﻪ ﻻﯾﻪ ﮐﻮﻣﻮﻟﻮس از ﻓﻮﻟﯿﮑﻮلﻫﺎي 2-8 ﻣﯿﻠﯽﻣﺘﺮي ﺗﺨﻤﺪانﻫﺎي ﮔﺎوي اﺧﺬ از ﺷﺪه از ﮐﺸﺘﺎرﮔﺎه ﭘﺲ از ﺳﻪ ﻣﺮﺗﺒﻪ ﺷﺴﺘﺸﻮ در ﻣﺤﯿﻂ ﺷﺴﺘﺸﻮ ﺑﻪ ﻣﺤﯿﻂﻫﺎي ﺑﻠﻮغ ﺑﺎ ﻏﻠﻈﺖﻫﺎي 0/5 و 1 و 2/5 و 5 ﻣﯿﮑﺮوﮔﺮم در ﻣﯿﻠﯽﻟﯿﺘﺮ از آﻟﮑﺎﻟﻮﺋﯿﺪ ﻫﺎرﻣﯿﻦ در دﻣﺎي 38/5 درﺟﻪ ﺳﺎﻧﺘﯽﮔﺮاد ﺑﺎ 5 درﺻﺪ دي اﮐﺴﯿﺪ ﮐﺮﺑﻦ ﺑﻪ ﻣﺪت 22-24 ﺳﺎﻋﺖ اﻧﮑﻮﺑﻪ ﺷﺪﻧﺪ. ﭘﺲ از ﮔﺬﺷﺖ زﻣﺎن اﻧﮑﻮﺑﺎﺳﯿﻮن ﮐﻮﻣﻮﻟﻮسﻫﺎي اﻃﺮاف ﺗﺨﻤﮏﻫﺎ ﺑﺮداﺷﺘﻪ ﺷﺪ و ﺑﺎﺑﺖ آزاد ﺷﺪن ﺟﺴﻤﮏ ﻗﻄﺒﯽ )ﺑﻠﻮغ( ﻣﻮرد ارزﯾﺎﺑﯽ ﻗﺮار ﮔﺮﻓﺖ. ﻧﺘﺎﯾﺞ ﺑﺎ اﺳﺘﻔﺎده از ﻧﺮماﻓﺰار SPSS ﺑﺮﻧﺎﻣﻪ ANOVA ﻣﻮرد ﺗﺠﺰﯾﻪ و ﺗﺤﻠﯿﻞ ﻗﺮار ﮔﺮﻓﺖ. ﻧﺘﺎﯾﺞ اﯾﻦ ﻣﻄﺎﻟﻌﻪ ﻧﺸﺎن دﻫﻨﺪه ﺗﺎﺛﯿﺮ ﻣﺜﺒﺖ آﻟﮑﺎﻟﻮﺋﯿﺪ ﻫﺎرﻣﯿﻦ در ﻏﻠﻈﺖ 1 ﻣﯿﮑﺮوﮔﺮم ﺑﺮ ﻣﯿﻠﯽﻟﯿﺘﺮ ﺑﻮده اﺳﺖ )0/05

In vitro maturation of oocytes is the technique that can reduce costs and eliminate the side effects of the use of gonadotropins for in vitro fertilization. Conducting research on the development and improvement of in vitro culture conditions for human oocyte maturation is very difficuly. The use domestic animals has provided the most advanced and best system to study the maturation of immature follicles in the laboratory. Due to the active ingredient in the harmine alkaloid with multiple pharmacological effects including anti-free radical, anti-inflammation and immune system, etc., the present study assesses the effects of harmine alkaloid on bovine oocyte maturation. In this study, at least three layers of cumulus oocytes aspirated from 2-8 mm follicles obtained from bovine ovaries from slaughterhouses after three washes in the washing environment were incubated for 22-24 hours in maturation environments with different concentrations of 0.5, 1, 2.5, and 5 µg/ml of the alkaloid harmine at a temperature of 38.5 ° C with 5% carbon dioxide. After a while, the cumulus incubation was removed from surrounding eggs were removed and the release of polar corpuscle (maturity) was assessed. Results were evaluated using SPSS and ANOVA. Research results showed the positive impact of the alkaloid harmine at 1 µg/ml (p <0.05) such that the maturity of the groups treated with doses of 0.5, 1, 2.5 and 5 micrograms per milliliter were 59.85, 72.65, 64.34, and 37.22 percent, respectively, which was 56.93 percent in the control group. Adding 1 µg/ml of alkaloids harmine led to the maturity environment increased the maturation of the bovine oocytes, which can be used as a supplement in oocyte maturation environments.