بررسي اثرات آنتي‌اكسيداني و مهاري عصاره دانه سويا (Glycine max) بر استيل كولين استراز و توليد نانوبيوفيبريل‌هاي آميلوئيدي

Investigation of antioxidant and Inhibitory properties of soybean seeds extract (Glycine max) on the acetylcholinesterase and the production of Amyloid Nano-biofibrils

ﺳﻮﯾﺎ )Glycine max( ﮔﯿﺎﻫﯽ از ﺧﺎﻧﻮاده ﺑﻘﻮﻻت و از ﻣﻬﻢ ﺗﺮﯾﻦ ﮔﯿﺎﻫﺎن روﻏﻨﯽ اﺳﺖ. داﻧﻪ ﺳﻮﯾﺎ ﺳﺮﺷﺎر از ﭘﺮوﺗﺌﯿﻦ و ﺗﺮﮐﯿﺒﺎت ﻓﯿﺘﻮﺷﯿﻤﯿﺎﯾﯽ ﻣﺨﺘﻠﻒ، ﺑﻪ وﯾﮋه اﯾﺰوﻓﻼون ﻫﺎ ﻣﯽﺑﺎﺷﺪ. ﻫﺪف از اﻧﺠﺎم اﯾﻦ ﺗﺤﻘﯿﻖ ﺑﺮرﺳﯽ اﺛﺮات آﻧﺘﯽاﮐﺴﯿﺪاﻧﯽ و ﻣﻬﺎري ﻋﺼﺎره ﻫﯿﺪرواﻟﮑﻠﯽ داﻧﻪ ﺳﻮﯾﺎ ﺑﺮ اﺳﺘﯿﻞ ﮐﻮﻟﯿﻦ اﺳﺘﺮاز و ﺗﻮﻟﯿﺪ ﻧﺎﻧﻮﺑﯿﻮﻓﯿﺒﺮﯾﻞ ﻫﺎي آﻣﯿﻠﻮﺋﯿﺪي در ﭘﺮوﺗﺌﯿﻦ آﻟﺒﻮﻣﯿﻦ ﺳﺮم ﮔﺎوي ﺑﻪﻋﻨﻮان ﯾﮏ ﭘﺮوﺗﺌﯿﻦ ﻣﺪل اﺳﺖ. اﺑﺘﺪا داﻧﻪ زردرﻧﮓ ﺳﻮﯾﺎ ﭘﻮدر ﺷﺪه، ﺳﭙﺲ ﻋﺼﺎره ﻫﯿﺪرواﻟﮑﻠﯽ آن ﺗﻬﯿﻪ ﮔﺮدﯾﺪ. ﺑﺎ روش ﮔﺎز ﮐﺮوﻣﺎﺗﻮﮔﺮاﻓﯽ ﺟﺮﻣﯽ، ﺗﺮﮐﯿﺒﺎت ﻋﺼﺎره ﺑﻪ دﺳﺖ آﻣﺪ. اﺛﺮات ﺿﺪآﻟﺰاﯾﻤﺮي ﺑﺎ ﺳﻨﺠﺶ ﻓﻌﺎﻟﯿﺖ آﻧﺘﯽ آﮐﺴﯿﺪاﻧﯽ ﺑﻪ روش DPPH، ﺑﺮرﺳﯽ ﻣﻬﺎر آﻧﺰﯾﻢ اﺳﺘﯿﻞ ﮐﻮﻟﯿﻦ اﺳﺘﺮاز ﺑﻪ روش اﻟﻤﻦ و ﺑﺮرﺳﯽ ﻣﻬﺎر ﺗﻮﻟﯿﺪ ﻧﺎﻧﻮﺑﯿﻮﻓﯿﺒﺮﯾﻞ ﻫﺎي آﻣﯿﻠﻮﺋﯿﺪي ﺑﻪ روش ﺟﺬب ﺳﻨﺠﯽ ﮐﻨﮕﻮرد اﻧﺠﺎم ﺷﺪ. از ﺗﺼﻮﯾﺮﺑﺮداري ﻣﯿﮑﺮوﺳﮑﻮپ اﻟﮑﺘﺮوﻧﯽ ﮔﺬاره ﺑﺮاي ﺗﺄﯾﯿﺪ ﺣﻀﻮر رﺷﺘﻪ ﻫﺎي آﻣﯿﻠﻮﺋﯿﺪي اﺳﺘﻔﺎده ﺷﺪ. ﺣﻀﻮر ﻣﺎده ﻣﺆﺛﺮه ﺟﻨﯿﺴﺘﺌﯿﻦ، ﺑﻪ ﻣﻘﺪار 2/97 درﺻﺪ و اﺳﯿﺪﻫﺎي ﭼﺮب ﭘﺎﻟﻤﯿﺘﯿﮏ و ﻟﯿﻨﻮﻟﺌﯿﮏ اﺳﯿﺪ ﺑﻪ ﺗﺮﺗﯿﺐ 3/11 و 10/69 درﺻﺪ در ﻋﺼﺎره داﻧﻪ زرد ﺳﻮﯾﺎ ﺗﺄﯾﯿﺪ ﺷﺪ. ﺑﺮرﺳﯽ ﻓﻌﺎﻟﯿﺖ آﻧﺘﯽ اﮐﺴﯿﺪاﻧﯽ ﻋﺼﺎره ﺳﻮﯾﺎ ﺑﺎ دوزﻫﺎي ﻣﺨﺘﻠﻒ ﻧﺸﺎن داد ﮐﻪ ﺑﺎ اﻓﺰاﯾﺶ ﻏﻠﻈﺖ ﻋﺼﺎره درﺻﺪ ﻓﻌﺎﻟﯿﺖ آﻧﺘﯽ اﮐﺴﯿﺪاﻧﯽ ﻧﯿﺰ اﻓﺰاﯾﺶ ﻣﯽﯾﺎﺑﺪ. ﺑﺮرﺳﯽ ﻣﻬﺎر آﻧﺰﯾﻢ اﺳﺘﯿﻞﮐﻮﻟﯿﻦاﺳﺘﺮاز ﺑﺎ اﺳﺘﻔﺎده از آﻧﺎﻟﻮگ ﺳﻮﺑﺴﺘﺮا ﺑﻪ ﻧﺎم اﺳﺘﯿﻞ ﺗﯿﻮﮐﻮﻟﯿﻦ ﯾﺪﯾﺪ )ATCI( ﻧﺸﺎن داد ﮐﻪ ﺑﺎ اﻓﺰاﯾﺶ ﻏﻠﻈﺖ ﻋﺼﺎره ﻣﯿﺰان ﻣﻬﺎر آﻧﺰﯾﻢ اﻓﺰاﯾﺶ ﻣﯽﯾﺎﺑﺪ. ﻣﯿﺰان ﻋﺼﺎره در ﻏﻠﻈﺖﻫﺎي ﺑﺎﻻ ﻣﻮﺟﺐ ﮐﺎﻫﺶ ﺗﻮﻟﯿﺪ ﻧﺎﻧﻮرﺷﺘﻪﻫﺎي آﻣﯿﻠﻮﺋﯿﺪي ﺷﺪ. اﯾﻦ ﻣﻄﻠﺐ ﺑﺎ ﺗﺼﺎوﯾﺮ ﻣﯿﮑﺮوﺳﮑﻮپ اﻟﮑﺘﺮوﻧﯽ ﻣﻮرد ﺗﺄﯾﯿﺪ ﻗﺮار ﮔﺮﻓﺖ. داﻧﻪ ﻫﺎي زرد ﺳﻮﯾﺎ، ﺑﺎ اﺛﺮات آﻧﺘﯽ اﮐﺴﯿﺪاﻧﯽ و ﻣﻬﺎري ﺧﻮﺑﯽ ﮐﻪ ﺑﺮ آﻧﺰﯾﻢ اﺳﺘﯿﻞﮐﻮﻟﯿﻦاﺳﺘﺮاز دارﻧﺪ، ﻣﯽ ﺗﻮاﻧﻨﺪ ﮐﺎﻧﺪﯾﺪاي ﻣﻨﺎﺳﺒﯽ ﺑﺮاي ﮐﺎﻫﺶ ﻋﻮارض ﺟﺎﻧﺒﯽ آﻟﺰاﯾﻤﺮ ﺑﺎﺷﻨﺪ.

Soybean (Glycine max) is a plant of the legume family and one of the most important oil plants. Soybean is rich in various proteins and phytochemicals, especially isoflavones. The aim of this study was to investigate the antioxidant and inhibitory effects of hydro-alcoholic extract of soybean seed on acetyl cholinesterase and production of amyloid nanobiofibrils in bovine serum albumin protein as a model protein. First, the yellow soybean seeds was powdered, then its hydro-alcoholic extract was prepared. The extracts composition were obtained by GC-MS spectroscopy. Anti-Alzheimer's effects were performed by measuring antioxidant activity by DPPH method, Inhibition of acetylcholinesterase by ellman method and inhibition of the production of amyloid nanobiofibrils by congored absorption method. Transmission electron microscopy was used to confirm the presence of amyloid fibrils. The presence of genistein active component in yellow soybean seed extract was confirmed by 2.97% and palmitic and linoleic acids by 3.11 and 10.69%, respectively. Investigation of antioxidant activity of soybean extract in different doses showed that with increasing the concentration of the extract, the percentage of antioxidant activity also increases. Investigation of acetylcholinesterase inhibition using a substrate analogue called acetylthiocholine iodide (ATCI) showed that with increasing the concentration of the extract, the amount of enzyme inhibition increases. The amounts of extract at high concentrations reduced the production of amyloid nanofibrils. This was confirmed by electron microscope images. Yellow soybeans, with their good antioxidant and inhibitory effects on the acetylcholinesterase enzyme, can be a good candidate for reducing the side effects of Alzheimer's.