اﺛﺮ ﻋﺼﺎره ﮔﯿﺎه ﻣﺸﮕﮏ ﺑﺮ ﺑﯿﺎن ﺑﺮﺧﯽ ژن ﻫﺎي ﻣﺮﺗﺒﻂ ﺑﺎ اﯾﻤﻨﯽ در ﻗﺰل آﻻي رﻧﮕﯿﻦ ﮐﻤﺎن

Effects of Ducrosia anethifolia Essential Oil on Some Immune Related Genes Expressions in Rainbow Trout

در ﺗﺤﻘﯿﻖ ﺣﺎﺿﺮ اﺛﺮ اﺳﺎﻧﺲ ﮔﯿﺎه ﻣﺸﮕﮏ ﺑﻪ ﻋﻨﻮان ﻣﺤﺮك اﯾﻤﻨﯽ در دوزﻫﺎي 0 ، 0/1 و 0/01، در ﻏﺬاي ﻣﺎﻫﯽ ﻗﺰل آﻻ در ﻣﺪت 50 روز ﺑﺮ ﺑﯿﺎن ژنﻫﺎي اﯾﻨﺘﺮﻟﻮﮐﯿﻦ (IL-1β) و ﻓﺎﮐﺘﻮر ﻧﮑﺮوز ﺗﻮﻣﻮري (TNF-α) در ﺑﺎﻓﺖﻫﺎي ﮐﻠﯿﻪ و ﻃﺤﺎل ﺑﺮرﺳﯽ ﺷﺪ. در روزﻫﺎي 10، 30 و 50 ﻧﻤﻮﻧﻪﺑﺮداري از ﺑﺎﻓﺖ ﮐﻠﯿﻪ و ﻃﺤﺎل ﻣﺎﻫﯽﻫﺎ ﺻﻮرت ﮔﺮﻓﺖ. ﭘﺲ از اﻧﺠﺎم ﻣﺮاﺣﻞ اﺳﺘﺨﺮاج RNA و ﺳﺎﺧﺖ ،cDNA، ﺑﯿﺎن ژنﻫﺎي IL-1β و TNF-α به روش Rt-qPCR ﺳﻨﺠﯿﺪه ﺷﺪ. دادهﻫﺎي ﺣﺎﺻﻞ ﺑﻪ روش CT∆∆ آﻧﺎﻟﯿﺰ ﺷﺪﻧﺪ و ﺳﭙﺲ اﺧﺘﻼف آﻣﺎري ﺗﯿﻤﺎرﻫﺎي ﻣﺨﺘﻠﻒ در روزﻫﺎي ﻣﺘﻔﺎوت ﺑﺎ اﺳﺘﻔﺎده از روﯾﻪ Mix و ﺑﻪ روش اﻧﺪازهﮔﯿﺮي ﺗﮑﺮارﺷﻮﻧﺪه در ﻧﺮم اﻓﺰار SAS ﻣﻘﺎﯾﺴﻪ ﮔﺮدﯾﺪ. ﻧﺘﺎﯾﺞ ﺣﺎﺻﻠﻪ ﻧﺸﺎن داد ﮐﻪ ﺑﯿﺸﺘﺮﯾﻦ ﺑﯿﺎن ﺑﺮاي ﻫﺮ دو ژن IL-1β و TNF-α در ﺑﺎﻓﺖ ﮐﻠﯿﻪ، ﻣﺮﺑﻮط ﺑﻪ ﺗﯿﻤﺎر 001/. ، 0 و روز 30 ﺑﻮده اﺳﺖ .و ﮐﻤﺘﺮﯾﻦ ﺑﯿﺎن ﺑﺮاي ژن IL-1β در تيمار 001/. روز 50 و ﺑﺮاي ژن TNF-α تيمار 0.1 در روز 50 مي باشد. درﻣﻮرد ﺑﺎﻓﺖ ﻃﺤﺎل،ﺑﯿﺸﺘﺮﯾﻦ ﺑﯿﺎن ﺑﺮاي IL-1β و TNF-α در روز 30 و به ترتيب در تيمارهاي 001/. و 0.1 ﺑﻮده اﺳﺖ. ﮐﻤﺘﺮﯾﻦ ﺑﯿﺎن اﻧﺪازهﮔﯿﺮي ﺷﺪه در اﯾﻦ ﺑﺎﻓﺖ، ﺑﺮاي ﺑﺮاي ﻫﺮ دو ژن IL-1β و TNF-αﻧﯿﺰ در روز 10 و ﺑﻪ ﺗﺮﺗﯿﺐ ﺗﯿﻤﺎرﻫﺎي 001/. و 0/1 ﺑﻮده اﺳﺖ. ﺑﻪ ﻃﻮر ﮐﻠﯽ ﻧﺘﺎﯾﺞ اﯾﻦ ﺗﺤﻘﯿﻖ ﻧﺸﺎن داد ﺗﻐﺬﯾﻪ ﻣﺎﻫﯿﺎن در ﻃﯽ ﺳﯽ روز ﺑﺎ دوزﻫﺎي ﻣﯿﺎﻧﻪ و ﭘﺎﯾﯿﻦ ﺑﯿﺸﺘﺮﯾﻦ ﺗﺎﺛﯿﺮ را در ﺑﯿﺎن ژﻧﻬﺎي ﻣﻮرد ﻣﻄﺎﻟﻌﻪ داﺷﺘﻪ اﺳﺖ.

The effect of moshgak (Ducrosia anethifolia) essential oil, as immunostimulant on the expression of IL-1β and TNF-α was studied in spleen and kidney of rainbow trout (Oncorhynchus mykiss). Fish were allocated in four different groups and fed on the diet containg essential oil at levels of 0, 0.1, 0.01 and 0.001% for 50 days. Tissue samples were taken on days 10, 30 and 50, and after RNA extraction and cDNA syntesis, the expession of IL-1β and TNF-α was studied by Rt-qPCR. Transcription data were analysed using ∆∆Ct. The significancy of means were analysed by repeated measurement method in Mix procedure using SAS software. Results showed that the highest expression of both IL-1β and TNF-α genes in kidney was observed on day 30 and the lowest expression for IL-1β and TNF-α was obtained on day 50 at doses 0.001 and 0.1%, respectively. In spleen, the highest expression of IL-1β and TNF-α was obtained on day 30 at doses 0.1 and 0.01%, respectively. While the lowest expression level of both genes was observed on day 10 at doses 0.001 and 0.1%, respectively. In conclusion, the results of this study showed that feeding of rainbow trout with low and moderate levels of moshgak essential oil for 30 days led to highest expression of studied immune related genes.