تاثير كاهش بيان گيرنده كموكايني CXCR3 بر اختلال عملكرد سلول هاي اندوتليال رگ

The Effect of Chemokine CXCR3 Receptor Downregulation on Vascular Endothelial Cell Dysfunction

آﺗﺮواﺳﮑﻠﺮوز ﯾﮏ ﺑﯿﻤﺎري ﻋﺮوﻗﯽ ﻣﺰﻣﻦ و ﻋﺎﻣﻞ اﺻﻠﯽ ﻣﺮگ و ﻣﯿﺮ در ﺳﺮاﺳﺮ ﺟﻬﺎن اﺳﺖ. اﺧﺘﻼل در ﻋﻤﻠﮑﺮد ﺳﻠﻮلﻫﺎي اﻧﺪوﺗﻠﯿﺎل ﻋﺎﻣﻞ ﻣﻬﻤﯽ در اﯾﺠﺎد آﺗﺮواﺳﮑﻠﺮوزﯾﺰ اﺳﺖ. اﻓﺰاﯾﺶ ﺑﯿﺎن ژنﻫﺎي ﺷﺎﺧﺺ ﭼﺴﺒﻨﺪﮔﯽ ﺳﻠﻮﻟﯽ و ﮐﺎﻫﺶ ﭘﺮوﺗﺌﯿﻦﻫﺎي ﻣﺘﺼﻞﮐﻨﻨﺪه ﺳﻠﻮلﻫﺎ ﻣﻨﺠﺮ ﺑﻪ ﻋﻤﻠﮑﺮد ﻏﯿﺮﻃﺒﯿﻌﯽ اﻧﺪوﺗﻠﯿﻮم ﻣﯽﺷﻮد. اﯾﻦ ﺗﻐﯿﯿﺮات ﻣﻮﻟﮑﻮﻟﯽ از ﻣﻬﻢﺗﺮﯾﻦ ﻧﺸﺎﻧﮕﺮﻫﺎي اﺧﺘﻼل در ﺳﻠﻮلﻫﺎي اﻧﺪوﺗﻠﯿﺎل و ﭘﯿﺸﺮﻓﺖ ﺑﯿﻤﺎري آﺗﺮواﺳﮑﻠﺮوز اﺳﺖ. CXCR3 ﯾﮏ ﮔﯿﺮﻧﺪه ﮐﻤﻮﮐﺎﯾﻨﯽ Gﭘﺮوﺗﺌﯿﻨﯽ اﺳﺖ ﮐﻪ ﺗﻮﺳﻂ ﺳﻠﻮلﻫﺎي اﻧﺪوﺗﻠﯿﺎل ﺑﯿﺎن ﻣﯽﺷﻮد. ﻧﻘﺶ ﮔﯿﺮﻧﺪه CXCR3 و ﻟﯿﮕﺎﻧﺪﻫﺎي آن در ﻋﻤﻠﮑﺮد ﺳﻠﻮلﻫﺎي اﻧﺪوﺗﻠﯿﺎﻟﯽ و اﯾﺠﺎد ﺑﯿﻤﺎريﻫﺎي ﻗﻠﺒﯽ ﻫﻨﻮز ﺑﻪدرﺳﺘﯽ ﺷﻨﺎﺧﺘﻪ ﻧﺸﺪه اﺳﺖ. در ﻣﻄﺎﻟﻌﻪ ﺣﺎﺿﺮ، ﺗﺎﺛﯿﺮ ﮐﺎﻫﺶ ﺑﯿﺎن ژن CXCR3 ﺑﺮ ﻣﯿﺰان ﺑﯿﺎن ﻧﺸﺎﻧﮕﺮﻫﺎي ﭼﺴﺒﻨﺪﮔﯽ )I-CAM-1 و V-CAM-1( و اﺗﺼﺎل ﻣﺤﮑﻢ (TJP1) ﺑﯿﻦ ﺳﻠﻮﻟﯽ ﻣﻮرد ﺑﺮرﺳﯽ ﻗﺮار ﮔﺮﻓﺖ. ﺑﻪﻣﻨﻈﻮر ﮐﺎﻫﺶ ﺑﯿﺎن ژن CXCR3 از DNAzyme ﺑﺮشدﻫﻨﺪه ﻋﻠﯿﻪ mRNA ژن CXCR3 اﺳﺘﻔﺎده ﺷﺪ. DNAzyme ﺗﻮﺳﻂ ﺗﻮرﺑﻮﻓﮑﺖ ﺑﻪ درون ﺳﻠﻮلﻫﺎي HUVEC ﺗﺮاﻧﺴﻔﮑﺖ ﺷﺪ. ﺑﻌﺪ از ﺗﺎﯾﯿﺪ ﮐﺎﻫﺶ ﺑﯿﺎن ژن CXCR3، اﺳﺘﺨﺮاج RNA و ﺳﻨﺘﺰ CDNA اﻧﺠﺎم ﺷﺪ و ﺳﭙﺲ ﺑﯿﺎن ﺗﺮاﻧﺴﮑﺮﯾﭙﺖ ژنﻫﺎي ﻫﺪف ﺗﻮﺳﻂ ﺗﮑﻨﯿﮏ RT-qPCR ﻣﻮرد ارزﯾﺎﺑﯽ ﻗﺮار ﮔﺮﻓﺖ. ﻧﺘﺎﯾﺞ ﻧﺸﺎن داد ﮐﻪ ﺳﻄﺢ ﺑﯿﺎن ICAM-1 و VCAM-1 ﺑﻪﻃﻮر ﻗﺎﺑﻞ ﺗﻮﺟﻬﯽ در ﺳﻠﻮلﻫﺎي ﺗﺮاﻧﺴﻔﮑﺖﺷﺪه در ﻣﻘﺎﯾﺴﻪ ﺑﺎ ﺳﻠﻮلﻫﺎي ﮐﻨﺘﺮل اﻓﺰاﯾﺶ ﯾﺎﻓﺖ، در ﺣﺎﻟﯽ ﮐﻪ ژن TJP1 ﺗﻐﯿﯿﺮ ﻗﺎﺑﻞ ﺗﻮﺟﻬﯽ ﻧﺸﺎن ﻧﺪاد. ﺑﻪ ﻧﻈﺮ ﻣﯽرﺳﺪ ﮐﻪ ﮐﺎﻫﺶ ﺑﯿﺎن ژن CXCR3 ﻣﯽﺗﻮاﻧﺪ زﻣﯿﻨﻪﺳﺎز اﯾﺠﺎد اﺧﺘﻼل در ﻋﻤﻠﮑﺮد ﺳﻠﻮلﻫﺎي اﻧﺪوﺗﻠﯿﺎﻟﯽ از ﻃﺮﯾﻖ اﻓﺰاﯾﺶ ﺑﯿﺎن ﻣﻮﻟﮑﻮلﻫﺎي ﭼﺴﺒﻨﺪﮔﯽ ﺷﻮد. ﺑﻨﺎﺑﺮاﯾﻦ، اﯾﻦ ﮔﯿﺮﻧﺪه ﻣﯽﺗﻮاﻧﺪ ﺑﻪﻋﻨﻮان ﯾﮏ ﻫﺪف ﻣﻮﻟﮑﻮﻟﯽ ﺑﺎﻟﻘﻮه ﺑﺮاي درك ﺑﻬﺘﺮي از ﻣﮑﺎﻧﯿﺰ م اﯾﺠﺎد ﺑﯿﻤﺎري آﺗﺮواﺳﮑﻠﺮوز در ﻧﻈﺮ ﮔﺮﻓﺘﻪ ﺷﻮد.

Atherosclerosis is a chronic vascular disease and remains the leading cause of death and morbidity worldwide. Endothelial dysfunction is an important factor in the progression of atherosclerotic disease. Increased expression of cell adhesion index genes and decreased cell-binding proteins lead to abnormal endothelial function. These molecular changes are one of the most important indicators of endothelial cell dysfunction and the progression of atherosclerosis. CXCR3 is a G-protein-coupled chemokine receptor expressed by endothelial cells. The role of the receptor CXCR3 and its ligands in endothelial cells and heart disease is not yet fully understood. In the current study, the effect of CXCR3 downregulation on the expression level of adhesion (I-CAM-1, V-CAM-1), tight junction (TJP1), related to endothelial dysfunction is evaluated. In order to reduce the expression of the CXCR3 gene, the RNA-cleaving DNAzyme was used against the mRNA of the CXCR3 gene. DNAzyme was transfused into HUVEC cells by TurboFectTM. After confirmation of decreased CXCR3 gene expression, RNA extraction and cDNA synthesis were performed and then the expression of markers was evaluated by RT-qPCR technique. The results depicted that the expression level of I-CAM-1 and V-CAM-1 were showed significant up-regulation in transfected cells compared with control cells, while the TJP1 gene was not showed significant change. It seems that reducing the CXCR3 gene expression could induce endothelial dysfunction through the change of adhesion markers genes expression. Therefore, this receptor can be considered as a potential molecular target for a better understanding of the mechanism of atherosclerosis.