تغييرات بياني ژن‌هاي كلاژن І و ІІІ در سلول‌هاي فيبروبلاست پوست انسان در اثر عصاره ريزجلبك Chlorella vulgaris و مقايسه آن با ويتامين C

Gene expression changes of collagen І and ІІІ in human skin fibroblast cells in effect of microalga Chlorella vulgaris extract and compared to vitamin C

ﭘﯿﺮي ﭘﻮﺳﺖ ﯾﮏ ﻓﺮآﯾﻨﺪ ﺑﯿﻮﻟﻮژﯾﮏ ﻧﺎﺷﯽ از ﮐﺎﻫﺶ ﺗﻮﻟﯿﺪ ﮐﻼژن و ﻧﯿﺰ اﻓﺰاﯾﺶ آﻧﺰﯾﻢﻫﺎي ﻣﺘﻌﺪد ﺷﺎﻣﻞ ﻣﺎﺗﺮﯾﮑﺲ ﻣﺘﺎﻟﻮﭘﺮوﺗﺌﯿﻨﺎزﻫﺎ )MMPS( ﺑﻮده ﮐﻪ اﯾﻦ آﻧﺰﯾﻢﻫﺎ ﻧﯿﺰ ﺑﺎﻋﺚ ﺗﺨﺮﯾﺐ ﮐﻼژن ﻣﯽﺷﻮﻧﺪ. Chlorella vulgaris ﯾﮏ رﯾﺰ ﺟﻠﺒﮏ درﯾﺎﯾﯽ ﻣﯽﺑﺎﺷﺪ و اﺛﺮات ﻣﻔﯿﺪش ﺑﺮ ﭘﻮﺳﺖ، آن را ﺑﻪ ﯾﮏ ﻣﺎده ﻣﻨﺎﺳﺐ ﺑﺮاي اﺳﺘﻔﺎده در ﻣﺤﺼﻮﻻت ﺿﺪ ﭘﯿﺮي ﺗﺒﺪﯾﻞ ﮐﺮده اﺳﺖ. در اﯾﻦ ﭘﮋوﻫﺶ، ﺗﺄﺛﯿﺮ ﻋﺼﺎره ﺟﻠﺒﮏ C. vulgaris در ﺗﻮﻟﯿﺪ ﮐﻼژن ﻧﻮع І و ІІІ در ﻣﻘﺎﯾﺴﻪ ﺑﺎ وﯾﺘﺎﻣﯿﻦ C در ﺳﻠﻮلﻫﺎي ﻓﯿﺒﺮوﺑﻼﺳﺖ ﭘﻮﺳﺖ )Hu02( در ﻣﺮﮐﺰ ﻣﻠﯽ ذﺧﺎﯾﺮ ژﻧﺘﯿﮑﯽ و زﯾﺴﺘﯽ اﯾﺮان در ﺷﻬﺮﯾﻮر 1397 ﺑﺮرﺳﯽ ﺷﺪ. ﻋﺼﺎرهﮔﯿﺮي از ﺟﻠﺒﮏ ﺑﻪ روش اوﻟﺘﺮاﺳﻮﻧﯿﮏ و ﻫﯿﺪروﻟﯿﺰ آﻧﺰﯾﻤﯽ اﻧﺠﺎم و ﻋﺼﺎره ﺣﺎﺻﻞ در ﻏﻠﻈﺖﻫﺎي ﻣﺨﺘﻠﻒ ﺑﺮاي ﺗﺄﺛﯿﺮ آن ﺑﺮ ﺑﯿﺎن ژن ﮐﻼژنﻫﺎي ﻧﻮع І وІІІ ﺑﺮ ﺳﻠﻮلﻫﺎي Hu02 اﺛﺮ داده ﺷﺪ. در اﺑﺘﺪا اﺛﺮ ﺳﻤﯿﺖ ﺳﻠﻮﻟﯽ آن ﺑﺮ ﺳﻠﻮل آزﻣﺎﯾﺶ و ﺑﺎ ﺗﺴﺖ MTT ﻣﺸﺨﺺ ﺷﺪ ﮐﻪ ﻋﺼﺎره ﻓﺎﻗﺪ اﺛﺮ ﺳﻤﯽ ﺑﺮاي ﺳﻠﻮلﻫﺎي Hu02 اﺳﺖ. ﻫﻤﭽﻨﯿﻦ ﺑﺎ اﺳﺘﻔﺎده از qRT-PCR ﺗﺄﯾﯿﺪ ﺷﺪ ﮐﻪ ﻋﺼﺎره ﺑﺎﻋﺚ اﻓﺰاﯾﺶ ﺑﯿﺎن ژن ﮐﻼژن ﻧﻮع І و ІІІ در ﻣﻘﺎﯾﺴﻪ ﺑﺎ ﮔﺮوه ﮐﻨﺘﺮل ﻣﯽﺷﻮد. ﻋﺼﺎره ﺟﻠﺒﮏ و وﯾﺘﺎﻣﯿﻦ C ﺑﺎ درﺻﺪ زﻧﺪهﻣﺎﻧﯽ )Viability( ﯾﮑﺴﺎن )97 درﺻﺪ( ﺑﺮاي ﺗﻌﯿﯿﻦ ﺗﺄﺛﯿﺮ ﻋﺼﺎره ﺑﺮ ﺑﯿﺎن ژنﻫﺎي ﮐﻼژن І و ІІІ در ﻣﻘﺎﯾﺴﻪ ﺑﺎ وﯾﺘﺎﻣﯿﻦ C اﺳﺘﻔﺎده ﺷﺪ. ﻋﺼﺎره ﺟﻠﺒﮏ و وﯾﺘﺎﻣﯿﻦ C ﺑﺎﻋﺚ اﻓﺰاﯾﺶ ﺑﯿﺎن ژن ﮐﻼژن ﻧﻮع І ﺑﻪ ﺗﺮﺗﯿﺐ ﺑﻪ ﻣﯿﺰان 3/14 ﺑﺮاﺑﺮ و 1/42 ﺑﺮاﺑﺮ ﺷﺪﻧﺪ. ﻋﺼﺎره ﺟﻠﺒﮏ ﺗﺄﺛﯿﺮ ﺑﯿﺸﺘﺮي در ﻣﻘﺎﯾﺴﻪ ﺑﺎ وﯾﺘﺎﻣﯿﻦ C ﺑﺮ ﺑﯿﺎن ژن ﮐﻼژن ﻧﻮع І و وﯾﺘﺎﻣﯿﻦ C ﺗﺄﺛﯿﺮ ﺑﯿﺸﺘﺮي در اﻓﺰاﯾﺶ ﺑﯿﺎن ژن ﮐﻼژن ﻧﻮع ІІІ در ﻣﻘﺎﯾﺴﻪ ﺑﺎ ﻋﺼﺎره ﺟﻠﺒﮏ داﺷﺖ و ﺑﺎﻋﺚ اﻓﺰاﯾﺶ ﺑﯿﺎن اﯾﻦ ژن ﺑﻪ ﻣﯿﺰان 2/12 ﺑﺮاﺑﺮ ﺷﺪ. درﺣﺎﻟﯽﮐﻪ اﯾﻦ ﻣﻘﺪار ﺑﺮاي ﻋﺼﺎره 1/14 ﺑﺮاﺑﺮ ﺑﻮد. اﯾﻦ ﯾﺎﻓﺘﻪﻫﺎ ﻧﺸﺎن داد ﮐﻪ ﻋﺼﺎره ﺟﻠﺒﮏ C. vulgaris و وﯾﺘﺎﻣﯿﻦ C ﺑﺎﻋﺚ ﺗﺤﺮﯾﮏ ﺳﻨﺘﺰ ﮐﻼژن در ﺳﻠﻮلﻫﺎي Hu02 ﻣﯽﺷﻮﻧﺪ و ﻣﯽﺗﻮاﻧﻨﺪ ﺟﺎﯾﮕﺰﯾﻦ ﻣﻨﺎﺳﺐ ﺑﺮاي اﻧﻮاع ﻣﻮاد ﺷﯿﻤﯿﺎﯾﯽ ﻣﻀﺮي ﺑﺎﺷﻨﺪ ﮐﻪ ﺑﺮاي ﮐﻼژن ﺳﺎزي در ﭘﻮﺳﺖ اﺳﺘﻔﺎده ﻣﯽﺷﻮﻧﺪ

Skin aging is a biological process that is due to the reduction of collagen production and increase of multiple enzymes, including matrix metalloproteinase (MMPS), which degrade collagen. Chlorella vulgaris is a marine microalga and its beneficial effects on the skin make it a proper ingredient to be used in anti-aging products. In this study, the effect of C. vulgaris extract comparing to vitamin C on types І and ІІІ collagen production in the human cell line Hu02 was investigated at the Iranian Biological Resource Center in September 2018. Chlorella was extracted using ultrasonication plus enzymatic hydrolysis, and Chlorella extract at different concentration was investigated for its effect on gene expression of collagen І and ІІІ in Hu02 cells. First, it was investigated whether alga extract induced cytotoxicity in Hu02 cells. MTT assay showed that extract was non-toxic to Hu02 cells. Using quantitative PCR, it was confirmed that extract increased the gene expression of types І and ІІІ collagen, comparing to the control group. Chlorella extract and vitamin C with a similar viability (97%) was used to investigate the effect of extract on gene expression of types І and ІІІ collagen, compared to vitamin C. Extract and vitamin C increased gene expression of type І collagen 3.14-fold and 1.42-fold, respectively. Alga extract had more effect on gene expression of type І collagen, compared to the vitamin C and vitamin C had more effect on gene expression of type ІІІ collagen, compared to the alga extract and increased its expression 2.12-fold, whereas this amount for the extract was 1.14 fold. These findings indicated that C. vulgaris extract and vitamin C induces collagen synthesis in Hu02 cells and could be proper alternative ingredients to the harmful chemicals used to make skin collagen.