ﻣﻘﺎﯾﺴﻪ اﺛﺮات ﺿﺪ ﻗﺎرﭼﯽ ﻋﺼﺎره ﻫﺎي آﺑﯽ و اﻟﮑﻠﯽ و اﺳﺎﻧﺲ ﭘﻮﻧﻪ ﺑﺎ داروي ﻓﻠﻮﮐﻮﻧﺎزول ﺑﺮ رﺷﺪ ﺳﻮﯾﻪ ﻫﺎي ﻗﺎرچ ﮐﺎﻧﺪﯾﺪا آﻟﺒﯿﮑﻨﺲ

In Vitro comparison of the efficacy of peppermint aqueous, alcoholic extract, essential oil and Fluconazole against Candida Strains

در ﺳﺎل ﻫﺎي اﺧﯿﺮ ﺷﯿﻮع ﺑﯿﻤﺎري ﻫﺎي ﻗﺎرچ ﻫﺎي ﻓﺮﺻﺖ ﻃﻠﺐ اﻓﺰاﯾﺶ ﭼﺸﻢ ﮔﯿﺮي ﯾﺎﻓﺘﻪ اﺳﺖ. ﮐﺎﻧﺪﯾﺪا آﻟﺒﯿﮑﻨﺲ ﯾﮑﯽ از ﭘﺎﺗﻮژن ﻫﺎي ﻗﺎرﭼﯽ ﻣﻌﻤﻮل اﺳﺖ ﮐﻪ ﻧﺴﺒﺖ ﺑﻪ داروﻫﺎي ﮔﺮوه آزول ﻣﻘﺎوم ﺷﺪه اﺳﺖ. در اﯾﻦ ﺗﺤﻘﯿﻖ ﺑﻪ ﻣﻘﺎﯾﺴﻪ اﺛﺮ ﺿﺪﻗﺎرﭼﯽ ﻋﺼﺎره آﺑﯽ و اﻟﮑﻠﯽ و اﺳﺎﻧﺲ ﮔﯿﺎه ﭘﻮﻧﻪ روي رﺷﺪ ﮐﺎﻧﺪﯾﺪا آﻟﺒﯿﮑﻨﺲ ﺑﺎ داروي ﻓﻠﻮﮐﻮﻧﺎزول ﭘﺮداﺧﺘﻪ ﺷﺪه اﺳﺖ. در اﯾﻦ ﻣﻄﺎﻟﻌﻪ 100 ﻧﻤﻮﻧﻪ ﻗﺎرﭼﯽ ﮐﺎﻧﺪﯾﺪاﯾﯽ از ﺑﯿﻤﺎران ﺗﻬﯿﻪ ﺷﺪ. ﺑﺎ اﻧﺠﺎم ﺗﺴﺖ ﻫﺎي ﺗﺸﺨﯿﺼﯽ ﻣﯿﮑﺮوﺳﮑﻮﭘﯽ، ﺑﯿﻮﺷﯿﻤﯿﺎﯾﯽ و آزﻣﺎﯾﺶ ﺟﺬب ﻗﻨﺪ ﺗﻮﺳﻂ ﮐﯿﺖ API20C ﻧﻤﻮﻧﻪ ﻫﺎي ﻗﺎرﭼﯽ ﮐﺎﻧﺪﯾﺪا آﻟﺒﯿﮑﻨﺲ ﺟﺪا ﺷﺪ. ﺳﭙﺲ روش ﻣﻮﻟﮑﻮﻟﻲ PCR-RFLP ﺑﺮاي ﺷﻨﺎﺳﺎﯾﻲ ﮔﻮﻧﻪ ﻫﺎي ﻣﺨﺘﻠﻒ ﻣﺨﻤﺮ ﮐﺎﻧﺪﯾﺪا و اﻓﺘﺮاق آن ﻫﺎ از ﯾﮑﺪﯾﮕﺮ )ﺑﺎ ﮐﺎﻧﺪﯾﺪا آﻟﺒﯿﮑﻨﺲ( ﻣﻮرد اﺳﺘﻔﺎده ﻗﺮار ﮔﺮﻓﺖ. ﻃﺒﻖ اﺳﺘﺎﻧﺪارد اﻧﺘﺸﺎر از دﯾﺴﮏ NCCLS، دﯾﺴﮏ ﻫﺎي 25 ﻣﯿﻠﯽ ﮔﺮﻣﯽ ﻓﻠﻮﮐﻮﻧﺎزول ﺗﻬﯿﻪ ﺷﺪ. اﺳﺘﺨﺮاج ﻋﺼﺎره ﺑﻪ روش ﭘﺮﮐﻮﻻﺳﯿﻮن و اﺳﺘﺨﺮاج اﺳﺎﻧﺲ ﺑﻪ روش ﺗﻘﻄﯿﺮ ﺑﺎ آب ﺻﻮرت ﮔﺮﻓﺖ. ﺳﻮﯾﻪ اﺳﺘﺎﻧﺪارد ﮐﺎﻧﺪﯾﺪاي ATCC14053 ﺟﻬﺖ ارزﯾﺎﺑﯽ ﮐﻨﺘﺮل ﮐﯿﻔﯽ ﮐﺎر اﺳﺘﻔﺎده ﺷﺪ. ﻣﻘﺎﯾﺴﻪ اﺛﺮ ﺿﺪﻗﺎرﭼﯽ ﻋﺼﺎره ﻫﺎي ﭘﻮﻧﻪ ﺑﻪ ﻫﻤﺮاه اﺳﺎﻧﺲ ﭘﻮﻧﻪ ﺑﺎ داروي ﻓﻠﻮﮐﻮﻧﺎزول، روي اﯾﻦ ﺟﺪاﯾﻪ ﻫﺎ ﺑﺎ دو روش دﯾﺴﮏ دﯾﻔﯿﻮژن و ﻣﯿﮑﺮوداﯾﻠﻮﺷﻦ اﻧﺠﺎم ﺷﺪ. ﺑﯿﺶ ﺗﺮﯾﻦ ﻓﺮاواﻧﯽ ﻧﺴﺒﯽ ﻣﺨﻤﺮ ﺟﺪا ﺷﺪه از ﻧﻤﻮﻧﻪ ﻫﺎي ﮐﺎﻧﺪﯾﺪاﯾﯽ ﻣﺮﺑﻮط ﺑﻪ ﮐﺎﻧﺪﯾﺪا آﻟﺒﯿﮑﻨﺲ ﺑﻮد ﮐﻪ در روش ﺗﺸﺨﯿﺺ ﺑﯿﻮﺷﯿﻤﺎﯾﯽ 35% و در روش PCR-RFLP ﻣﯿﺰان 38% از ﮐﻞ ﻧﻤﻮﻧﻪ ﻫﺎ را ﺷﺎﻣﻞ ﺷﺪ. ﻣﯿﺎﻧﮕﯿﻦ ﻗﻄﺮ ﻫﺎﻟﻪ ﻫﺎي ﻋﺪم رﺷﺪ ﺑﺮاي ﻋﺼﺎره ﻫﺎي آﺑﯽ و اﻟﮑﻠﯽ ﭘﻮﻧﻪ 1، اﺳﺎﻧﺲ ﭘﻮﻧﻪ 23/4 و ﺑﺮاي ﻓﻠﻮﮐﻮﻧﺎزول 26/16 ﻣﯿﻠﯽ ﻣﺘﺮ ﺑﻮد ﮐﻪ ﻧﺸﺎن دﻫﻨﺪه ﻋﻤﻠﮑﺮد ﺿﺪ ﻗﺎرﭼﯽ ﺑﻬﺘﺮ ﻓﻠﻮﮐﻮﻧﺎزول و ﺗﻔﺎوت ﻣﻌﻨﯽ دار آن ﻧﺴﺒﺖ ﺑﻪ اﺳﺎﻧﺲ ﭘﻮﻧﻪ ﺑﻮد )0.05 ≤P (. ﻣﯿﺰان ﺣﺪاﻗﻞ ﻏﻠﻈﺖ ﻋﺼﺎره اﻟﮑﻠﯽ و آﺑﯽ ﭘﻮﻧﻪ و ﻓﻠﻮﮐﻮﻧﺎزول ﺑﻪ ﺗﺮﺗﯿﺐ 4 و 8 و 8 ﻣﯿﻠﯽ ﮔﺮم ﺑﺮ ﻣﯿﻠﯽ ﻟﯿﺘﺮ ﺑﺮاي MIC و 8 و 16 و 8 ﻣﯿﻠﯽ ﮔﺮم ﺑﺮ ﻣﯿﻠﯽ ﻟﯿﺘﺮ ﺑﺮاي MFC ﺑﻮد. ﻣﯿﺰان اﻧﺤﺮاف ﻣﻌﯿﺎر ﻋﺼﺎره اﻟﮑﻠﯽ ﭘﻮﻧﻪ در ﺗﺴﺖ MFCﺑﺮاﺑﺮ ﺑﺎ 3/55 ﺑﻮد؛ در ﺣﺎﻟﯽ ﮐﻪ ﺑﺮاي ﻓﻠﻮﮐﻮﻧﺎزول 3/68 ﺑﻮد. ﺑﺎ ﺗﻮﺟﻪ ﺑﻪ اﯾﻨﮑﻪ از ﻟﺤﺎظ آﻣﺎري ﻫﺮ ﭼﻪ اﻧﺤﺮاف ﻣﻌﯿﺎر ﮐﻤﺘﺮ ﺑﺎﺷﺪ، ﻋﻤﻠﮑﺮد ﺑﻬﺘﺮ اﺳﺖ؛ از اﯾﻦ رو ﻋﺼﺎره اﻟﮑﻠﯽ ﭘﻮﻧﻪ ﻋﻤﻠﮑﺮد ﺑﻬﺘﺮي ﻧﺴﺒﺖ ﺑﻪ ﻓﻠﻮﮐﻮﻧﺎزول داﺷﺖ. اﯾﻦ ﻧﺘﺎﯾﺞ ﺑﯿﺎﻧﮕﺮ ﻋﻤﻠﮑﺮد ﺑﻬﺘﺮ و ﺗﻔﺎوت ﻣﻌﻨﯽ دار ﻋﺼﺎره اﻟﮑﻠﯽ ﭘﻮﻧﻪ ﻧﺴﺒﺖ ﺑﻪ ﻓﻠﻮﮐﻮﻧﺎزول ﻣﯽ ﺑﺎﺷﺪ )0.05 ≤P(. ﺑﺎ ﺗﻮﺟﻪ ﺑﻪ ﻧﺘﺎﯾﺞ ﺣﺎﺻﻞ از ﺗﺤﻘﯿﻖ ﻓﻮق، ﺑﻪ ﻧﻈﺮ ﻣﯽ رﺳﺪ ﻋﺼﺎره اﻟﮑﻠﯽ ﮔﯿﺎه ﭘﻮﻧﻪ ﻣﯽ ﺗﻮاﻧﺪ ﺑﻪ ﻋﻨﻮان ﯾﮏ ﮐﺎﻧﺪﯾﺪ داروﯾﯽ ﻣﻨﺎﺳﺐ ﺑﺎ ﻋﻤﻠﮑﺮدي ﺿﺪ ﻗﺎرﭼﯽ ﺑﻬﺘﺮ از ﻓﻠﻮﮐﻮﻧﺎزول در ﺻﻨﺎﯾﻊ داروﺳﺎزي ﻣﻮرد اﺳﺘﻔﺎده ﻗﺮار ﮔﯿﺮد.

In recent years, the prevalence of opportunistic fungal diseases has increased significantly. Candida albicans is a common fungal pathogen that has become resistant to azole drugs. In this study, the antifungal effect of aqueous and alcoholic extracts and peppermint essential oil on the growth of Candida albicans with fluconazole was investigated. In this study, 100 candidate fungal samples were prepared from patients. Candida albicans fungal samples were isolated by microscopic, biochemical, and glucose uptake tests by API20C kit. Then, the molecular PCR-RFLP method was used to identify different species of Candida yeast and differentiate them from each other (with Candida albicans). According to the NCCLS disc release standard, 25 mg fluconazole discs were prepared. The extract was extracted by percolation and the essential oil was extracted by water distillation. The standard strain of the ATCC14053 candidate was used to evaluate the quality control of the work. Comparison of the antifungal effect of Mentha pulegium extracts with peppermint essential oil wi was performed on these isolates by disk diffusion and microdilution methods. The highest relative -RFLP method. The mean diameter of non-growth halos for aqueous and alcoholic extracts of Mentha pulegium was 1, Mentha pulegium essential oil was 23.4 mm and for fluconazole was 26.16 mm, which showed better antifungal performance of fluconazole Mentha pulegium concentrations of alcoholic and aqueous extracts of mg/ml for MIC and 8, 16, and 8 mg/ml for MFC, respectively. The standard deviation of ct in the MFC test was 3.55; While for fluconazole, it was 3.68. Given that statistically the smaller the standard deviation, the better the performance; Therefore, pulegium alcoholic extract had a better performance than fluconazole. These results indicate better performance and significant difference between Mentha pulegium alcoholic extract than fluconazole (P ≤ 0.05). According to the results of the above study, it seems that the alcoholic extract of Mentha pulegium can be used as a suitable drug candidate with better antifungal performance than fluconazole in the pharmaceutical industry.