ﻣﻘﺎﯾﺴﻪ ﺗﺄﺛﯿﺮ ﻋﺼﺎره ﺳﯿﺮ و ﺟﻨﺘﺎﻣﺎﯾﺴﯿﻦ روي ﺑﯿﺎن ژن ﺑﯿﻮﻓﯿﻠﻢ icaA در Staphylococcus aureus

Comparison of the effect of garlic extract and gentamicin on icaA biofilm gene expression in Staphylococcus aureus

Staphylococcus aureus ﯾﮑﯽ از ﺷﺎﯾﻊ ﺗﺮﯾﻦ ﺑﯿﻤﺎرﮔﺮﻫﺎي ﺑﺎﮐﺘﺮﯾﺎﯾﯽ اﺳﺖ. ﺟﻨﺘﺎﻣﺎﯾﺴﯿﻦ ﯾﮑﯽ از آﻧﺘﯽ ﺑﯿﻮﺗﯿﮏ ﻫﺎﯾﯽ اﺳﺖ ﮐﻪ ﺑﺮاي درﻣﺎن ﻋﻔﻮﻧﺖ ﻫﺎي اﺳﺘﺎﻓﯿﻠﻮﮐﻮﮐﻮﺳﯽ ﻣﻮﺛﺮ اﺳﺖ وﻟﯽ ﻣﺘﺎﺳﻔﺎﻧﻪ ﺑﻪ دﻟﯿﻞ ﺗﻮﻟﯿﺪ ﺑﯿﻮﻓﯿﻠﻢ، ﺗﺄﺛﯿﺮ اﯾﻦ آﻧﺘﯽﺑﯿﻮﺗﯿﮏ ﮔﺎﻫﯽ ﺑﺎ ﻣﺸﮑﻞ روﺑﻪرو ﻣﯽ ﮔﺮدد. در اﯾﻦ ﭘﮋوﻫﺶ، ﺗﺄﺛﯿﺮ ﻋﺼﺎره ﺳﯿﺮ و ﻣﻘﺎﯾﺴﻪ آن ﺑﺎ آﻧﺘﯽ ﺑﯿﻮﺗﯿﮏ ﺟﻨﺘﺎﻣﺎﯾﺴﯿﻦ روي ﺳﻪ ﺳﻮﯾﻪ از ﺑﺎﮐﺘﺮي ﻫﺎي S. aureus ﺑﺮ اﺳﺎس روش آﻧﺘﯽ ﺑﯿﻮﮔﺮام و آزﻣﻮن MIC ﻣﻮرد ﺳﻨﺠﺶ ﻗﺮار ﮔﺮﻓﺖ ﮐﻪ ﻧﺘﺎﯾﺞ آن ﺗﺄﯾﯿﺪﮐﻨﻨﺪه ﺗﺄﺛﯿﺮ ﺿﺪ اﺳﺘﺎﻓﯿﻠﻮﮐﻮﮐﯽ ﻋﺼﺎره ﺳﯿﺮ و اﺛﺮ ﻫﻢ اﻓﺰاﯾﯽ آن ﺑﺎ آﻧﺘﯽ ﺑﯿﻮﺗﯿﮏ ﺟﻨﺘﺎﻣﺎﯾﺴﯿﻦ ﺑﻮد. ﺑﺮ اﺳﺎس آزﻣﻮن ﻓﻨﻮﺗﯿﭙﯽ، اﺛﺮ ﻋﺼﺎره ﺳﯿﺮ و ﺟﻨﺘﺎﻣﺎﯾﺴﯿﻦ ﺑﻪﺗﻨﻬﺎﯾﯽ و ﺗﻮأم ﺑﺎ ﻫﻢ ﺑﺮ ﮐﺎﻫﺶ ﻣﯿﺰان ﺗﻮﻟﯿﺪ ﺑﯿﻮﻓﯿﻠﻢ در اﯾﻦ ﺑﺎﮐﺘﺮي ارزﯾﺎﺑﯽ و اﺛﺒﺎت ﺷﺪ. وﺟﻮد ژن icaA در ﺳﻮﯾﻪ ﻫﺎي ﻣﻮرد ﻣﻄﺎﻟﻌﻪ ﺗﻮﺳﻂ PCR ﺑﻪ اﺛﺒﺎت رﺳﯿﺪ و ﺑﺎ اﺳﺘﻔﺎده از ﺗﮑﻨﯿﮏ Real time PCR، ﻣﯿﺰان ﺑﯿﺎن ژن icaA ﺗﺤﺖ ﺗﯿﻤﺎر ﻋﺼﺎره ﺳﯿﺮ، ﺟﻨﺘﺎﻣﺎﯾﺴﯿﻦ و ﺗﯿﻤﺎر ﺗﻮأﻣﺎن ﺳﯿﺮ و ﺟﻨﺘﺎﻣﺎﯾﺴﯿﻦ ﻧﺴﺒﺖ ﺑﻪ ﻧﻤﻮﻧﻪ ﻫﺎي ﺑﺪون ﺗﯿﻤﺎر ﻣﻮرد ﻣﻄﺎﻟﻌﻪ ﻗﺮار ﮔﺮﻓﺖ. در ﻣﻮرد ﺳﻮﯾﻪ اﺳﺘﺎﻧﺪارد، ﺟﻨﺘﺎﻣﺎﯾﺴﯿﻦ و ﻋﺼﺎره ﺳﯿﺮ ﺑﻪ ﺗﺮﺗﯿﺐ ﺑﺎﻋﺚ ﮐﺎﻫﺶ 59 درﺻﺪي و 64 درﺻﺪي ﺑﯿﺎن ژن icaA ﺷﺪﻧﺪ و اﺳﺘﻔﺎده ﺗﻮأم اﯾﻦ دو ﻣﺎده، ﺑﯿﺎن اﯾﻦ ژن را ﺑﻪ ﺣﺪود 25 درﺻﺪ ﻧﻤﻮﻧﻪ ﮐﻨﺘﺮل ﮐﺎﻫﺶ داد ﯾﻌﻨﯽ ﻣﻨﺠﺮ ﺑﻪ ﮐﺎﻫﺶ 75 درﺻﺪي ﺑﯿﺎن اﯾﻦ ژن ﮔﺮدﯾﺪ. اﯾﻦ ﺗﯿﻤﺎرﻫﺎ در ﻣﻮرد ﺳﻮﯾﻪ ﻫﺎي ﺑﯿﻤﺎرﮔﺮ ﻧﯿﺰ ﺗﺄﺛﯿﺮ زﯾﺎدي ﺑﺮ ﮐﺎﻫﺶ ﺑﯿﺎن icaA ﻧﺸﺎن دادﻧﺪ ﺑﻪ ﻃﻮري ﮐﻪ اﺳﺘﻔﺎده ﻫﻢزﻣﺎن ﻋﺼﺎره ﺳﯿﺮ و ﺟﻨﺘﺎﻣﺎﯾﺴﯿﻦ ﺗﻮاﻧﺴﺖ ﺑﯿﺎن اﯾﻦ ژن را ﺑﻪ 11 درﺻﺪ ﻧﻤﻮﻧﻪ ﺑﺪون ﺗﯿﻤﺎر ﮐﺎﻫﺶ دﻫﺪ و ﻣﻨﺠﺮ ﺑﻪ ﮐﺎﻫﺶ 89 درﺻﺪي ﺑﯿﺎن icaA ﮔﺮدد. ﻧﺘﺎﯾﺞ ﺗﻤﺎم اﯾﻦ آزﻣﻮن ﻫﺎ ﺣﺎﮐﯽ از ﺗﺄﺛﯿﺮ ﻣﻄﻠﻮب ﻋﺼﺎره ﺳﯿﺮ در ﻣﻘﺎﺑﻞ ﺑﺎﮐﺘﺮي S. aureus ﺑﻮد.

Staphylococcus aureus is one of the most common bacterial pathogens. Gentamicin is one of the antibiotics that is effective in treating staphylococcal infections, but unfortunately due to biofilms, the effect of this antibiotic is sometimes confronted with the problem. In this study, the effect of garlic extract and its comparison with gentamicin antibiotic on three strains of S. aureus bacteria was evaluated based on antibiogram method and MIC test, the results of which confirm the anti-staphylococcal effect of garlic extract and its synergistic effect with gentamicin antibiotic. Based on phenotypic test, the effect of garlic extract and gentamicin alone and together on reducing the production of biofilm in this bacterium was evaluated and proven. The presence of icaA gene in the studied strains was confirmed by PCR and using Real time PCR technique, the expression of icaA gene under the treatment of garlic extract, gentamicin and the combined treatment of garlic and gentamicin compared to the untreated samples was studied. For the standard strain, gentamicin and garlic extract reduced icaA gene expression by 59% and 64%, respectively, and combined use of these two substances reduced the expression of this gene by about 25% compared to the control sample. In other words, it led to a 75% reduction in the expression of this gene. In the case of pathogenic strains, these treatments also showed a significant effect on reducing icaA expression, so that the simultaneous use of garlic extract and gentamicin could reduce the expression of this gene to 11% of the untreated sample. In other words, it leads to an 89% reduction in icaA expression. The results of all these tests showed the favorable effect of garlic extract against S. aureus.