بررسي نقش miR-124-3p در بيان ژن هاي عصبي و غير عصبي در سلولهاي بنيادي فوليكول مو

Study on the role of miR-124-3p on the expression of neuronal and nonneuronal genes in the hair follicle stem cells

ﺳﺎﺑﻘﻪ و ﻫﺪف: ﺳﻠﻮلﻫﺎي ﺑﻨﯿﺎدي ﻓﻮﻟﯿﮑﻮل ﻣﻮ، ﻗﺎﺑﻠﯿﺖ ﺗﻤﺎﯾﺰ ﺑﻪ ﺳﻠﻮلﻫﺎي ﻧﻮروﻧﯽ و ﮔﻠﯿﺎل را دارﻧﺪ. ﻣﯿﮑﺮوآرﻧﺎﻫﺎي ﻣﺨﺘﻠﻒ، ﺗﮑﺜﯿﺮ و ﺗﻤﺎﯾﺰ ﺳﻠﻮلﻫﺎي ﺑﻨﯿﺎدي ﻓﻮﻟﯿﮑﻮل ﻣﻮ را ﺗﻨﻈﯿﻢ ﻣﯽﮐﻨﻨﺪ. ﺑﻨﺎﺑﺮاﯾﻦ ﻫﺪف از اﯾﻦ ﻣﻄﺎﻟﻌﻪ، ﺑﺮرﺳﯽ ﻧﻘﺶ دﻗﯿﻖ miR-124-3p ﺑﺮ روي ﺑﯿﺎن ژنﻫﺎي ﻏﯿﺮﻋﺼﺒﯽ ))RCOR2 (CoREST ) ، NR2E1 (Tlx) ، CCND1 (cyclinD1( و ژنﻫﺎي ﻋﺼﺒﯽ )MAP2 ، DLK1 و )NeuN( Rbfox3( ﻣﯽﺑﺎﺷﺪ. ﻣﻮاد و روشﻫﺎ: ﻣﻄﺎﻟﻌﻪ ﺣﺎﺿﺮ ﺑﻪﺻﻮرت ﺗﺠﺮﺑﯽ ﺑﺮ روي ﺳﻠﻮلﻫﺎي ﺑﻨﯿﺎدي ﺟﺪاﺷﺪه از ﻧﺎﺣﯿﻪ ﺑﺎﻟﺞ ﻓﻮﻟﯿﮑﻮلﻫﺎي ﻣﻮي ﻟﺐ ﻓﻮﻗﺎﻧﯽ ﻣﻮش اﻧﺠﺎم ﺷﺪ. ﺳﻠﻮلﻫﺎي ﺑﻨﯿﺎدي ﻓﻮﻟﯿﮑﻮل ﻣﻮ ﺑﻪ ﺳﻠﻮلﻫﺎي ﻧﻮروﻧﯽ ﺗﻤﺎﯾﺰ داده ﺷﺪﻧﺪ. ﻫﻤﭽﻨﯿﻦ، ﺑﺨﺸﯽ از ﻣﯿﮑﺮوآرﻧﺎﻫﺎي ﺗﻘﻠﯿﺪﮐﻨﻨﺪه )Mimic( و ﻣﻬﺎرﮐﻨﻨﺪه )Inhibitor( ﻣﺮﺑﻮط ﺑﻪ miR-124-3p ﺑﻪ ﺳﻠﻮلﻫﺎي ﺑﻨﯿﺎدي ﻓﻮﻟﯿﮑﻮل ﻣﻮ اﻧﺘﻘﺎل داده ﺷﺪﻧﺪ. ﺳﻄﻮح ﺑﯿﺎن ژنﻫﺎي ﻏﯿﺮﻋﺼﺒﯽ و ﻋﺼﺒﯽ ﺗﻮﺳﻂ روش real-time PCR ﻣﻮرد ارزﯾﺎﺑﯽ ﻗﺮار ﮔﺮﻓﺖ. ﻧﺘﺎﯾﺞ: ﻧﺘﺎﯾﺞ ﺑﻪدﺳﺖآﻣﺪه ﻧﺸﺎن داد اﻟﻘﺎي ﺗﻤﺎﯾﺰ ﻧﻮروﻧﯽ و اﺳﺘﻔﺎده از ﻣﯿﮑﺮوآرﻧﺎﻫﺎي ﺗﻘﻠﯿﺪﮐﻨﻨﺪه miR-124-3p، ﺳﻄﺢ ﺑﯿﺎن ژنﻫﺎي ﻋﺼﺒﯽ Rbfox3 ، MAP2 و DLK1 را ﺑﻪﺻﻮرت ﻣﻌﻨﯽداري اﻓﺰاﯾﺶ ﻣﯽدﻫﺪ؛ وﻟﯽ اﻧﺘﻘﺎل ﻣﻬﺎرﮐﻨﻨﺪه miR-124-3p ﺑﻪ داﺧﻞ ﺳﻠﻮلﻫﺎ، ﺑﺎﻋﺚ اﻓﺰاﯾﺶ ﻣﻌﻨﯽدار 82/71 و 962/07 ﺑﺮاﺑﺮي در ﺳﻄﻮح mRNAي ژنﻫﺎي ﻏﯿﺮﻋﺼﺒﯽ CCND1 و NR2E1 ﮔﺮدﯾﺪ. ﻧﺘﯿﺠﻪﮔﯿﺮي: اﯾﻦ ﯾﺎﻓﺘﻪﻫﺎ ﻧﺸﺎن ﻣﯽدﻫﺪ ﮐﻪ miR-124-3p ﺑﺎ ﻣﻬﺎر ژنﻫﺎي ﻏﯿﺮﻋﺼﺒﯽ CCND1 و NR2E1 ﻣﻤﮑﻦ اﺳﺖ ﻣﻨﺠﺮ ﺑﻪ اﻓﺰاﯾﺶ ﺗﻤﺎﯾﺰ ﻋﺼﺒﯽ ﺳﻠﻮلﻫﺎي ﺑﻨﯿﺎدي ﻓﻮﻟﯿﮑﻮل ﻣﻮ ﺷﻮد.

Hair follicle stem cells (HFSCs) can be differentiated into neurons and glial cells. Different miRNAs regulate the proliferation and differentiation of HFSCs. So, this study aimed to evaluate the exact role of miR-124-3p on the expression of non-neuronal genes (CCND1 (cyclinD1), Tlx )NR2E1) and RCOR2 (CoREST)( and neuronal genes (DLK1, MAP2, and NeuN (Rbfox 3)). Material and Methods: This experimental study was performed on the stem cells isolated from the bulge region of mouse vibrissa hair follicles. The hair follicle stem cells differentiated into neuronal cells. Then, the cells transfected with miR-124-3p mimic and inhibitor. The mRNA levels of non-neuronal and neural genes were detected by real-time PCR (RT-PCR). Results: The results showed that neural induction and miR-124-3p mimic transfection significantly increased mRNA expression levels of neuronal genes MAP2, Rbfox3 and DLK1, but transfection with the miR-124-3p inhibitor increased mRNA level of non-neural genes CCND1 (82.71) and NR2E1 (962.07), respectively. Conclusion: These results show that miR-124-3p may promote neuronal differentiation of HFSCs by inhibiting non-neural genes CCND1 and NR2E1