بررسي اثر نانوذرات نقره بر بيان ژن هاي اگزوتوكسين A و S در سودوموناس آئروژينوزا

The effect of silver nanoparticles on the expression of exotoxin A and S genes in Pseudomonas aeruginosa

ﭼﮑﯿﺪه ﻫﺪف: ﺳﻮدوﻣﻮﻧﺎس آﺋﺮوژﯾﻨﻮزا ﯾﮑﯽ از ﻣﻬﻢ ﺗﺮﯾﻦ ﻋﻮاﻣﻞ ﻋﻔﻮﻧﺖ ﻫﺎي ﺑﯿﻤﺎرﺳﺘﺎﻧﯽ اﺳﺖ و ژن ﻫﺎي اﮔﺰوﺗﻮﮐﺴﯿﻦ در ﺑﺮوز اﯾﻦ ﻋﻔﻮﻧﺖ داراي اﻫﻤﯿﺖ ﻣﯽ ﺑﺎﺷﺪ. از ﺳﻮﯾﯽ ﺑﺎ ﭘﯿﺸﺮﻓﺖ ﻋﻠﻢ ﻓﻨﺎوري ﻧﺎﻧﻮ، اﻣﺮوزه اﺳﺘﻔﺎده از ﻧﺎﻧﻮذرات ﺑﻪ ﻃﻮر ﮔﺴﺘﺮده اي در زﻣﯿﻨﻪ درﻣﺎﻧﯽ و داروﯾﯽ ﺑﻮﯾﮋه ﺑﺮ ﻋﻠﯿﻪ ﻣﯿﮑﺮوب ﻫﺎ ﻣﻮرد ﺗﻮﺟﻪ ﻗﺮار ﮔﺮﻓﺘﻪ اﺳﺖ. ﻟﺬا، ﺗﺤﻘﯿﻖ ﺣﺎﺿﺮ ﺑﺎ ﻫﺪف ﺑﺮرﺳﯽ ﺗﺎﺛﯿﺮ ﻧﺎﻧﻮذرات ﻧﻘﺮه AgNPs)( ﺑﺮ ﺑﯿﺎن ژن ﻫﺎي اﮔﺰوﺗﻮﮐﺴﯿﻦ A و S ﺳﻮدوﻣﻮﻧﺎس آﺋﺮوژﯾﻨﻮزا اﻧﺠﺎم ﺷﺪه اﺳﺖ.ﻣﻮاد و روش ﻫﺎ: در اﯾﻦ ﻣﻄﺎﻟﻌﻪ ﺗﺠﺮﺑﯽ، ﺣﺪاﻗﻞ ﻏﻠﻈﺖ ﻣﻬﺎر رﺷﺪ ﻧﺎﻧﻮذرات ﻧﻘﺮه ﺑﺎ روش ﻣﯿﮑﺮوداﯾﻠﻮﺷﻦ ﺗﻌﯿﯿﻦ ﺷﺪ. اﺑﺘﺪا ﮐﻢ ﺗﺮﯾﻦ ﻏﻠﻈﺖ ﻣﻬﺎرﮐﻨﻨﺪه رﺷﺪ ﺑﺎﮐﺘﺮي ﺑﺎ ﻧﺎﻧﻮذرات ﻧﻘﺮه ﺗﻌﯿﯿﻦ ﮔﺮدﯾﺪه، ﺳﭙﺲ در ﻏﻠﻈﺖ ﭘﺎﯾﯿﻦ ﺗﺮ از ﮐﻢ ﺗﺮﯾﻦ ﻏﻠﻈﺖ ﻣﻬﺎر رﺷﺪ، ﻣﯿﺰان ﺑﯿﺎن ژن ﻫﺎي اﮔﺰوﺗﻮﮐﺴﯿﻦ A و S ﺳﻮدوﻣﻮﻧﺎس آﺋﺮوژﯾﻨﻮزا ﺑﺎ روش Real Time PCR ﻣﻮرد ﺑﺮرﺳﯽ ﻗﺮار ﮔﺮﻓﺖ. داده ﻫﺎي ﺑﻪ دﺳﺖ آﻣﺪه ﺑﺎ اﺳﺘﻔﺎده از ANONA ﯾﮏ ﻃﺮﻓﻪ، ﺗﺴﺖ ﺗﻮﮐﯽ و p value ﮐﻢ ﺗﺮ از 0/05 ارزﯾﺎﺑﯽ ﺷﺪ. ﻧﺘﺎﯾﺞ ﻧﺎﻧﻮذرات ﻧﻘﺮه ﺑﺎ ﻏﻠﻈﺖ 62/5 ﻣﯿﮑﺮوﮔﺮم ﺑﺮ ﻣﯿﻠﯽ ﻟﯿﺘﺮ، اﺛﺮات ﻣﻬﺎر رﺷﺪ ﺑﺮ ﺑﺎﮐﺘﺮي ﺳﻮدوﻣﻮﻧﺎس آﺋﺮوژﯾﻨﻮزا را داﺷﺘﻪ، ﻫﻤﭽﻨﯿﻦ در ﻏﻠﻈﺖ 31/25 ﺑﯿﺎن ژن ﻫﺎي اﮔﺰوﺗﻮﮐﺴﯿﻦ A و S ﺳﻮدوﻣﻮﻧﺎس آﺋﺮوژﯾﻨﻮزا ﺑﻪ ﻧﺴﺒﺖ ﻣﻌﻨﯽ دار ﮐﺎﻫﺶ ﯾﺎﻓﺖ )P<0/05(. ﻧﺘﺎﯾﺞ: ﻧﺎﻧﻮذرات ﻧﻘﺮه ﺑﺎ ﻏﻠﻈﺖ 62/5 ﻣﯿﮑﺮوﮔﺮم ﺑﺮ ﻣﯿﻠﯽ ﻟﯿﺘﺮ، اﺛﺮات ﻣﻬﺎر رﺷﺪ ﺑﺮ ﺑﺎﮐﺘﺮي ﺳﻮدوﻣﻮﻧﺎس آﺋﺮوژﯾﻨﻮزا را داﺷﺘﻪ، ﻫﻤﭽﻨﯿﻦ در ﻏﻠﻈﺖ 31/25 ﺑﯿﺎن ژن ﻫﺎي اﮔﺰوﺗﻮﮐﺴﯿﻦ A و S ﺳﻮدوﻣﻮﻧﺎس آﺋﺮوژﯾﻨﻮزا ﺑﻪ ﻧﺴﺒﺖ ﻣﻌﻨﯽ دار ﮐﺎﻫﺶ ﯾﺎﻓﺖ )P<0/05(. ﻧﺘﯿﺠﻪ ﮔﯿﺮي: ﻧﺘﺎﯾﺞ ﺣﺎﺻﻞ از اﯾﻦ ﻣﻄﺎﻟﻌﻪ ﻧﺸﺎن ﻣﯽ دﻫﺪ ﮐﻪ ﻏﻠﻈﺖ ﭘﺎﯾﯿﻦ ﺗﺮ )31/25 ﻣﯿﮑﺮوﮔﺮم ﺑﺮ ﻣﯿﻠﯽ ﻟﯿﺘﺮ( از ﻏﻠﻈﺖ ﻣﻬﺎر رﺷﺪ )62/5 ﻣﯿﮑﺮوﮔﺮم ﺑﺮ ﻣﯿﻠﯽ ﻟﯿﺘﺮ(، ﻣﯽ ﺗﻮاﻧﺪ ﺑﺎﻋﺚ ﮐﺎﻫﺶ ﺑﯿﺎن ژن ﻫﺎي اﮔﺰوﺗﻮﮐﺴﯿﻦ A و S ﺳﻮدوﻣﻮﻧﺎس آﺋﺮوژﯾﻨﻮزا ﺷﻮد.

Pseudomonas aeruginosa is one of the most important causes of nosocomial infections and exotoxin genes are important in the development of this infection.On the other hand, with the advancement of nanotechnology, today, the use of nanoparticles has been widely considered in the field of medicine and medicine, especially against microbes. The aim of the present study was to investigate the effect of silver nanoparticles (AgNPs) on the expression of Pseudomonas aeruginosa exotoxin A and S genes. Methods: In this experimental study, the minimum growth inhibition concentration of silver nanoparticles was determined by microdilution method. First, the lowest concentration of bacterial growth inhibitor is determined with silver nanoparticles. Then, at a concentration lower than the lowest growth rate, the expression of Pseudomonas aeruginosa exotoxin A and S genes was examined by Real time PCR. Data were analyzed using one-way ANOVA, Tukey test and P value less than 0.05. Findings: Silver nanoparticles with a concentration of 62.5 μg / ml had growth inhibitory effects on Pseudomonas aeruginosa. Also, at a concentration of 31.25, the expression of Pseudomonas aeruginosa exotoxin A and S genes was significantly reduced (P<0.05). Conclusion: The results of this study show that lower concentrations (31.25 μg/ml) than growth inhibition concentrations (62.5μg/ml) can reduce the expression of Pseudomonas aeruginosa exotoxin A and S genes.