بررسي تأثير داروي L-Dopa ليپوزومي و اگزوزوم‌هاي مشتق از سلول‌هاي SH-Sy5y بر بقاي نورون‌هاي دوپاميرنژيك از طريق تنظيم بيان ژن‌هاي Bdnf و Dat

Evaluation of the Effect of Liposomal L-Dopa and Exosomes Derived from SH-Sy5y Cells on the Survival of Dopaminergic Neurons by Regulating the Expression of Bdnf and Dat Genes

ﺑﯿﻤﺎري ﭘﺎرﮐﯿﻨﺴﻮن ﯾﮏ ﻧﺎﻫﻨﺠﺎري ﻧﻮرودژﻧﺮاﺗﯿﻮ ﭘﯿﺶ روﻧﺪه ﻣﯽﺑﺎﺷﺪ ﮐﻪ در ﻃﯽ آن ﻧﻮرونﻫﺎي دوﭘﺎﻣﯿﻨﺮژﯾﮏ در ﻧﺎﺣﯿﻪ ﺟﺴﻢ ﺳﯿﺎه ﺑﻪ ﺗﺪرﯾﺞ از ﺑﯿﻦ ﻣﯽروﻧﺪ. اﺧﯿﺮاً، روشﻫﺎي درﻣﺎﻧﯽ ﺗﺮﮐﯿﺒﯽ ﺑﻪ ﻋﻨﻮان روﯾﮑﺮدﻫﺎي درﻣﺎﻧﯽ ﻣﺆﺛﺮ ﺑﺴﯿﺎر ﻣﻮرد ﺗﻮﺟﻪ ﻗﺮار ﮔﺮﻓﺘﻪاﻧﺪ. در اﯾﻦ ﻣﻄﺎﻟﻌﻪ، اﺛﺮات ﺑﺎﻟﻘﻮه اﮔﺰوزومﻫﺎي ﺟﺪا ﺷﺪه از ﺳﻠﻮلﻫﺎي SH-Sy5y و داروي L-Dopa ﻟﯿﭙﻮزوﻣﯽ ﺑﺮ ﺷﺮاﯾﻂ ﭘﺎﺗﻮﻟﻮژﯾﮑﯽ ﻣﻮشﻫﺎي ﺻﺤﺮاﯾﯽ ﻣﺒﺘﻼ ﺑﻪ ﭘﺎرﮐﯿﻨﺴﻮن ﺑﺮرﺳﯽ ﮔﺮدﯾﺪ. ﻣﻮاد و روشﻫﺎ: 25 ﺳﺮ ﻣﻮش ﺻﺤﺮاﯾﯽ ﻧﺮ ﻧﮋاد وﯾﺴﺘﺎر در ﭘﻨﺞ ﮔﺮوه ﻃﺒﻘﻪﺑﻨﺪي ﺷﺪﻧﺪ. ﻣﺪل ﭘﺎرﮐﯿﻨﺴﻮن از ﻃﺮﯾﻖ ﺗﺰرﯾﻖ ﺗﺮﮐﯿﺐ 6-OHDA در ﻧﺎﺣﯿﻪ ﺟﺴﻢ ﺳﯿﺎه ﺳﻤﺖ راﺳﺖ ﻣﻐﺰ اﯾﺠﺎد ﺷﺪ. اﮔﺰوزومﻫﺎي ﺟﺪاﺳﺎزي ﺷﺪه از ﺳﻠﻮلﻫﺎي SH-Sy5y ﺑﻪ ﺗﻨﻬﺎﯾﯽ و ﻫﻤﺮاه ﺑﺎ داروي L-Dopa ﻟﯿﭙﻮزوﻣﯽ ﺷﺪه ﺑﻪ ﻣﺪلﻫﺎي ﺑﯿﻤﺎر ﺗﺰرﯾﻖ ﺷﺪﻧﺪ. اﺛﺮات ﺗﯿﻤﺎرﻫﺎي ﺻﻮرت ﮔﺮﻓﺘﻪ از ﻃﺮﯾﻖ اﻧﺠﺎم ﺗﺴﺖ رﻓﺘﺎري، ﺑﺮرﺳﯽﻫﺎي ﺑﺎﻓﺖﺷﻨﺎﺳﯽ و ﺑﺮرﺳﯽ ﺗﻐﯿﯿﺮات ﺑﯿﺎن دو ژن Bdnf و Dat ﺑﻪ روش real-time PCR ﺑﺮرﺳﯽ ﮔﺮدﯾﺪﻧﺪ. ﻧﺘﺎﯾﺞ: ﺗﯿﻤﺎر ﻫﻤﺰﻣﺎن ﻣﺪلﻫﺎي ﺑﯿﻤﺎر ﺑﺎ اﮔﺰوزوم و داروي L-Dopa ﻟﯿﭙﻮزوﻣﯽ ﺷﺪه ﺑﺎﻋﺚ اﻓﺰاﯾﺶ ﺟﻤﻌﯿﺖ ﻧﻮرونﻫﺎي دوﭘﺎﻣﯿﺮﻧﮋﯾﮏ در ﻧﺎﺣﯿﻪ ﺟﺴﻢ ﺳﯿﺎه ﻣﻐﺰ ﮔﺮدﯾﺪ. ﻫﻤﭽﻨﯿﻦ در ﻣﺪلﻫﺎي ﺗﯿﻤﺎر ﺷﺪه ﺑﺎ ﻫﺮ دو اﮔﺰوزوم و داروي L-Dopa ﻟﯿﭙﻮزوﻣﯽ ﺷﺪه اﻓﺰاﯾﺶ ﻣﻌﻨﺎدار ﺳﻄﺢ ﺑﯿﺎن ژنﻫﺎي Bdnf و Dat ﻣﺸﺎﻫﺪه ﺷﺪ در ﺣﺎﻟﯿﮑﻪ اﻓﺰاﯾﺶ ﺑﯿﺎن ﻣﺸﺎﻫﺪه ﺷﺪه در ﺗﯿﻤﺎرﻫﺎي ﺟﺪاﮔﺎﻧﻪ از ﻧﻈﺮ آﻣﺎري ﻣﻌﻨﺎدار ﻧﺒﻮدﻧﺪ )0/05>P (. ﻧﺘﯿﺠﻪﮔﯿﺮي: ﻧﺘﺎﯾﺞ ﻣﻄﺎﻟﻌﻪ ﺣﺎﺿﺮ ﻧﺸﺎن دادﻧﺪ ﮐﻪ اﺳﺘﻔﺎده از داروي L-Dopa ﺑﻪ ﻫﻤﺮاه اﮔﺰوزمﻫﺎي ﺳﻠﻮلﻫﺎي SH-Sy5y اﺛﺮات ﺳﯿﻨﺮژﯾﮏ ﻣﺜﺒﺘﯽ ﺑﺮ اﻓﺰاﯾﺶ ﺑﯿﺎن ژنﻫﺎي Bdnf و Dat داﺷﺘﻪ و ﺑﺎﻋﺚ اﻟﻘﺎي ﺗﻤﺎﯾﺰ ﺳﻠﻮلﻫﺎي ﭘﯿﺶ ﺳﺎز ﻋﺼﺒﯽ ﺑﻪ ﻧﻮرونﻫﺎي دوﭘﺎﻣﯿﻨﺮژﯾﮏ و در ﻧﺘﯿﺠﻪ اﻓﺰاﯾﺶ ﺟﻤﻌﯿﺖ و ﺑﻘﺎي آﻧﻬﺎ در ﻧﺎﺣﯿﻪ ﺟﺴﻢ ﺳﯿﺎه ﻣﻐﺰ ﻣﯽﮔﺮدد.

Parkinson's disease is a progressive neurodegenerative complication in which dopaminergic neurons located in substrantia nigra are gradually lost. Currently, the combined therapy strategies received more attentions as potential therapeutic approaches for Parkinson's disease. This study aimed to evaluate the potential effects of exosomes released from SH-Sy5y cells and liposomal L-dopa drug on Parkinson's rat models. Methods: Twenty five male Wistar albino rats, in 5 groups, were included in this study. Parkinson's rat models were induced through microinjection of 6-OHDA (2.5 mg/mL) into the right substrantia nigra. The exosomes released from SH-Sy5y cell line were isolated and administrated alone or in combination with liposomal L-Dopa to the defined model groups. The effects of the treatments were evaluated by behavioral testing, histological studies and expression changes of bdnf and dat genes by real-time PCR. Results: The rats in groups received the combined liposomal L-Dopa and exosome treatment and liposomal L-Dopa alone showed a significant improvement in their movement abilities. Also, in the models treated with both exosomes and liposomal L-Dopa drug, a significant increase in the expression level of Bdnf and Dat genes was observed, while the increase in expression observed in separate treatments was not statistically significant (P>0.05). Conclusion: The results of the present study showed that the use of L-Dopa with exosomes of SH-Sy5y cells had positive synergistic effects on increasing the expression of Bdnf and Dat genes and induced differentiation of neuronal progenitor cells into dopaminergic neurons and thus increased their population and survival is in the area.