اﺛﺮ ﺳﻄﻮح ﻣﺨﺘﻠﻒ ﮔﻠﻮﺗﺎﺗﯿﻮن درون ﭘﻮﺷﺎﻧﯽ ﺷﺪه ﺑﺮ ﮐﯿﻔﯿﺖ اﻧﺠﻤﺎد اﺳﭙﺮم ﮔﺎو

Effect of different levels of encapsulated glutathione on cryopreservation of bull sperm

ﻫﺪف از اﯾﻦ ﻣﻄﺎﻟﻌﻪ، ارزﯾﺎﺑﯽ اﺛﺮ ﺳﻄﻮح ﻣﺨﺘﻠﻒ ﮔﻠﻮﺗﺎﺗﯿﻮن اﺣﯿﺎ در رﻗﯿﻖﮐﻨﻨﺪه ﺑﺮ ﮐﯿﻔﯿﺖ اﺳﭙﺮم ﮔﺎو ﭘﺲ از ﻓﺮاﯾﻨﺪ اﻧﺠﻤﺎد و ذوب ﺑﻮد. در ﭘﮋوﻫﺶ ﺣﺎﺿﺮ، رﻗﯿﻖﮐﻨﻨﺪه ﺣﺎوي ﻧﺎﻧﻮﻟﯿﭙﻮزومﻫﺎي ﻏﻨﯽ ﺷﺪه ﺑﺎ ﭼﻬﺎر ﺳﻄﺢ ﺻﻔﺮ، 2/5 ،1 و 5 ﻣﯿﻠﯽ ﻣﻮﻻر آﻧﺘﯽ اﮐﺴﯿﺪان ﮔﻠﻮﺗﺎﺗﯿﻮن ﺑﻮد. ﻟﯿﭙﻮزومﻫﺎي ﺣﺎوي ﮔﻠﻮﺗﺎﺗﯿﻮن ﺑﺎ اﺳﺘﻔﺎده از روش ﻓﯿﻠﻢ ﻧﺎزك ﺗﻬﯿﻪ ﺷﺪ. اﻧﺪازه ذرات ﺑﺎ اﺳﺘﻔﺎده از دﺳﺘﮕﺎه ﺳﻮﻧﯿﮑﺎﺗﻮر ﺑﻪ اﺑﻌﺎد ﻧﺎﻧﻮ ﮐﺎﻫﺶ ﯾﺎﻓﺖ. ﭼﻬﻞ اﻧﺰال ﻃﯽ ﺷﺶ ﻫﻔﺘﻪ از ﺷﺶ راس ﮔﺎو ﻧﺮ ﻫﻠﺸﺘﺎﯾﻦ ﺟﻤﻊآوري و ﻣﻮرد اﻧﺠﻤﺎد ﻗﺮار ﮔﺮﻓﺖ. ﺻﻔﺎت ﻣﻮرد ارزﯾﺎﺑﯽ ﭘﺲ از اﻧﺠﻤﺎد و ذوب ﺷﺎﻣﻞ، ﻓﺮاﺳﻨﺠﻪﻫﺎي ﺟﻨﺒﺎﯾﯽ، ﻓﻌﺎﻟﯿﺖ ﻏﺸﺎ و ﯾﮑﭙﺎرﭼﮕﯽ ﻏﺸﺎ و رﯾﺨﺖﺷﻨﺎﺧﺘﯽ اﺳﭙﺮم ﺑﻮد. ﻧﺘﺎﯾﺞ اﯾﻦ ﻣﻄﺎﻟﻌﻪ ﻧﺸﺎن داد ﮐﻪ اﺳﺘﻔﺎده از 1 ﻣﯿﻠﯽ ﻣﻮﻻر ﮔﻠﻮﺗﺎﺗﯿﻮن ﺟﻨﺒﺎﯾﯽ ﮐﻞ و ﺟﻨﺒﺎﯾﯽ ﭘﯿﺸﺮوﻧﺪه را ﺑﻪ ﻃﻮر ﻣﻌﻨﯽ داري اﻓﺰاﯾﺶ ﻣﯽ دﻫﺪ )47/5±1/7 و 32/7±2 درﺻﺪ(، اﮔﺮﭼﻪ، ﺗﻔﺎوت ﻣﻌﻨﯽ داري ﺑﺎ ﻏﻠﻈﺖ 2/5 ﻣﯿﻠﯽ ﻣﻮﻻر ﻧﺪاﺷﺖ )45/0±1/7 و 26/2±2 درﺻﺪ(. ﻫﻤﭽﻨﯿﻦ، ﻏﻠﻈﺖ 2/5 ﻣﯿﻠﯽ ﻣﻮﻻر ﺑﺎﻻﺗﺮﯾﻦ ﻣﯿﺰان ﯾﮑﭙﺎرﭼﮕﯽ و ﺳﻼﻣﺖ ﻏﺸﺎ را ﻧﺸﺎن داد، اﻣﺎ ﺗﻔﺎوت ﻣﻌﻨﯽ داري ﺑﺎ ﻏﻠﻈﺖ 1 ﻣﯿﻠﯽ ﻣﻮﻻر ﻧﺸﺎن ﻧﺪاد. ﺑﺎ اﯾﻦ ﺣﺎل، 5 ﻣﯿﻠﯽﻣﻮﻻر ﮔﻠﻮﺗﺎﺗﯿﻮن درون ﭘﻮﺷﺎﻧﯽ ﺷﺪه ﺳﺒﺐ ﮐﺎﻫﺶ ﻓﺮاﺳﻨﺠﻪﻫﺎي ﮐﯿﻔﯽ اﺳﭙﺮم ﺷﺪ. ﺑﻨﺎﺑﺮاﯾﻦ، اﺳﺘﻔﺎده از ﺳﻄﺢ 1 ﻣﯿﻠﯽ ﻣﻮﻻر ﮔﻠﻮﺗﺎﺗﯿﻮن ﺑﻪ ﺷﮑﻞ درونﭘﻮﺷﺎﻧﯽ ﺷﺪه در رﻗﯿﻖﮐﻨﻨﺪه ﻣﯽﺗﻮاﻧﺪ ﺣﻔﺎﻇﺖ ﺑﻬﺘﺮي از اﺳﭙﺮم ﮔﺎو ﻧﺴﺒﺖ ﺑﻪ ﺳﺎﯾﺮ ﺳﻄﻮح ﺑﻪ ﻋﻤﻞ آورد.

This study was designed to evaluate the effects of encapsulated reduced glutathione added to extender on the quality of bull sperm after freezing and thawing process. Extender contains lecithin nanoliposomes supplemented with different concentrations of glutathione (0, 1, 2.5 and 5 Mm concentration). The supplemented nanoliposomes with glutathione were prepared by thin-film preparation method. The particle size decreased to nanometer by sonication. 40 ejaculations were collected from 6 Holstein bulls and frozen for 6 weeks. Evaluated characteristics after freezing and thawing process were kinetic parameters (CASA), membrane activity (HOST), membrane integrity (eosin-nigrosin) and morphology of sperm ( Hancock's solution). Results showed that level 1 mM significantly increased the total and progressive motility (47.5±1.7, 32.7±2 %). Although, there was no significant difference with level 2.5 mM glutathione (45.0±1.7, 26.2±2 %). Also, the concentration of 2.5 mM showed the highest value of cell membrane integrity and cell membrane functionality but did not differ with the concentration of 1mM. However, supplementation of nanoliposomes with 5 mM glutathione reduced the quality of post-thaw sperm. Thus, level 1 M encapsulated glutathione can provide better protection compare to the other level of that for bull sperm.