بررسي متابونوميكس اثر داروي آرتميزينين روي فاز ايستائي انگل ليشمانيا ماژور سويه فردلين در شرايط آزمايش گاهي

The metabonomic changes of leishmania major, s promastigotes (fredlin strain) after in vitro artemisinin treatment at stationary phase

ﺳﺎﺑﻘﻪ و ﻫﺪف: ﺑﺮ اﺳﺎس ﺗﻘﺴﯿﻢ ﺑﻨﺪي ﺳﺎزﻣﺎن ﺑﻬﺪاﺷﺖ ﺟﻬﺎﻧﯽ (WHO)، ﺑﯿﻤﺎري ﻟﯿﺸﻤﺎﻧﯿﺎزﯾﺲ از ﺟﻤﻠﻪ ﺷـﺶ ﺑﯿﻤـﺎري ﻣﻬـﻢ ﻣﻨﻄﻘﻪ ﺣﺎره اي ﻣﺤﺴﻮب ﻣﯽ ﮔﺮدد. ﻫﻢ ﭼﻨﯿﻦ اﯾﻦ ﺑﯿﻤﺎري در ﺧﺎورﻣﯿﺎﻧﻪ و ﮐﺸﻮرﻫﺎي ﻣﺪﯾﺘﺮاﻧﻪ اي ﻧﯿﺰ ﺷﺎﯾﻊ ﻣﯽ ﺑﺎﺷﺪ. اﻫﻤﯿﺖ ﮐﻨﺘـﺮل اﯾﻦ ﺑﯿﻤﺎري، ﺑﻪ دﻟﯿﻞ اﻓﺰاﯾﺶ ﻣﻮارد ﻣﺸﺎﻫﺪه ﺷﺪه در ﺑﯿﻤﺎران داراي ﻧﻘﺺ ﺳﯿﺴﺘﻢ اﯾﻤﻨﯽ از ﺟﻤﻠﻪ اﯾﺪز ﻣﯽ ﺑﺎﺷﺪ و ﺑﺎ ﺗﻮﺟﻪ ﺑـﻪ ﻧﯿـﺎز ﻣﺒﺮم ﺑﻪ داروﻫﺎي ﺟﺪﯾﺪ ﺑﺎ ﻋﻮارض ﮐﻢ ﺗـﺮ ﺑـﺮاي ﮐﻨﺘـﺮل و درﻣـﺎن ﺑﯿﻤـﺎري، ﺗـﺎﺛﯿﺮ داروي آرﺗﻤﯿـﺰﯾﻨﯿﻦ ﺑـﺮ روي ﻣﺘﺎﺑﻮﻟﯿـﺖ ﻫـﺎي ﭘﺮوﻣﺎﺳﺘﯿﮕﻮت ﻫﺎي اﻧﮕﻞ ﻟﯿﺸﻤﺎﻧﯿﺎ ﻣﺎژور (ﺳﻮش ﻓﺮدﻟﯿﻦ) در ﺷﺮاﯾﻂ آزﻣﺎﯾﺶ ﮔﺎﻫﯽ در اﻧﺴﺘﯿﺘﻮ ﭘﺎﺳﺘﻮر اﯾﺮان ﺑﺮرﺳﯽ ﮔﺮدﯾﺪ. ﻣﻮاد و روش ﻫﺎ: ﭘﺲ از ﺗﻌﯿﯿﻦ IC50 داروي آرﺗﻤﯿﺰﯾﻨﯿﻦ ﺑﺮ روي اﻧﮕﻞ ﻟﯿﺸﻤﺎﻧﯿﺎ؛ ﻣﺘﺎﺑﻮﻟﯿﺖ ﻫﺎي اﻧﮕﻞ در دو ﮔﺮوه ﮐﻨﺘﺮل (ﺑـﺪون درﯾﺎﻓﺖ دارو) و ﮔﺮوه ﻣﻮرد آزﻣﺎﯾﺶ (درﯾﺎﻓﺖ ﮐﻨﻨﺪه دوز ﻣﺸﺨﺺ از دارو) ﺑـﺎ اﺳـﺘﻔﺎده از روش ﻃﯿـﻒ ﺳـﻨﺠﯽ ﻣﻐﻨـﺎﻃﯿﺲ ﻫﺴـﺘﻪ HNMR ﻣﻮرد ﺑﺮرﺳﯽ ﻗﺮار ﮔﺮﻓﺖ. ﯾﺎﻓﺘﻪ ﻫﺎ: ﻧﺘﺎﯾﺞ ﻣﻄﺎﻟﻌﺎت ﻣﺎ ﻧﺸﺎن ﻣﯽدﻫﺪ ﮐﻪ ﺗﻐﯿﯿﺮات ﻣﺘﺎﺑﻮﻟﯿﺴﻢ ﻓﺎز اﯾﺴﺘﺎﯾﯽ ﭘﺮوﻣﺎﺳﺘﯿﮕﻮت ﻟﯿﺸـﻤﺎﻧﯿﺎ ﺳـﻮش ﻓـﺮدﻟﯿﻦ در دو ﮔﺮوه ﮐﻨﺘﺮل و ﮔﺮوه ﺗﺤﺖ اﺛﺮ داروي آرﺗﻤﯿﺰﯾﻨﯿﻦ، از ﺑﯿﻦ 41 ﻣﺘﺎﺑﻮﻟﯿﺖ ﭼﺮﺧﻪ ﻣﺘﺎﺑﻮﻟﯿﺴﻤﯽ ﮔﺎﻻﮐﺘﻮز، ﺗﻌﺪاد 5 ﻣﺘﺎﺑﻮﻟﯿﺖ ﺑـﯿﺶ ﺗـﺮﯾﻦ ﺗﻐﯿﯿﺮات را ﻧﺸﺎن دادﻧﺪ، ﻟﺬا ﻣﺘﺎﺑﻮﻟﯿﺖ ﻫﺎي ﻣﺴﯿﺮ ﮔﺎﻻﮐﺘﻮز رﺗﺒﻪ اول ﺗﻐﯿﯿـﺮات را دارد و ﺳـﭙﺲ ﻣﺘﺎﺑﻮﻟﯿـﺖ ﻫـﺎي ﻣﺴﯿﺮ ﺑﯿﻮﺳـﻨﺘﺰ اﺳﻔﻨﮕﻮﻟﯿﭙﯿﺪ و ﺑﯿﻮﺳﻨﺘﺰ واﻟﯿﻦ، ﻟﻮﺳﯿﻦ واﯾﺰوﻟﻮﺳﯿﻦ در درﺟﻪ ﺑﻌﺪي اﻫﻤﯿﺖ ﻗﺮار ﻣﯽ ﮔﯿﺮﻧﺪ. ﻧﺘﯿﺠﻪ ﮔﯿﺮي: داروي آرﺗﻤﯿﺰﯾﻨﯿﻦ در ﺷﺮاﯾﻂ آزﻣﺎﯾﺶ ﮔﺎﻫﯽ ﺑﺎ ﺗﻐﯿﯿﺮ در ﻣﺘﺎﺑﻮﻟﯿﺖ ﻫﺎي ﻣﺮﺑﻮط ﺑﻪ ﭼﺮﺧﻪ ﻫﺎي ﻣﺘﺎﺑﻮﻟﯿﺴﻤﯽ ﻣﺨﺘﻠﻒ از ﺟﻤﻠﻪ ﻣﺴﯿﺮ ﻣﺘﺎﺑﻮﻟﯿﺴﻤﯽ ﮔﺎﻻﮐﺘﻮز، ﻣﺴﯿﺮ ﺑﯿﻮﺳﻨﺘﺰ اﺳﻔﻨﮕﻮﻟﯿﭙﯿﺪ و ﻫﻢ ﭼﻨﯿﻦ ﻣﺴﯿﺮ ﺑﯿﻮﺳﻨﺘﺰ واﻟﯿﻦ، ﻟﻮﺳﯿﻦ و اﯾﺰوﻟﻮﺳﯿﻦ ﻣﻨﺠـﺮ ﺑـﻪ ﺗﻮﻗﻒ ﻓﻌﺎﻟﯿﺖ اﻧﮕﻞ ﻟﯿﺸﻤﺎﻧﯿﺎ ﻣﺎژور ﺳﻮﯾﻪ ﻓﺮدﻟﯿﻦ ﮔﺮدﯾﺪ. ﻃﯿﻒ ﻫـﺎي ﺣﺎﺻـﻠﻪ از آزﻣـﻮن HNMR ﺗﻮﺳـﻂ ﻧـﺮم اﻓـﺰار آﻣـﺎري ﮐﻤﻮﻣﺘﺮﯾﮑﺲ، اﺧﺘﻼف ﻣﻌﻨﯽداري را ﺑﯿﻦ ﻣﺘﺎﺑﻮﻟﯿﺖ ﻫﺎي ﮔﺮوه درﯾﺎﻓﺖ ﮐﻨﻨﺪه داروي آرﺗﻤﯿﺰﯾﻨﯿﻦ و ﮔﺮوه ﺑﺪون درﯾﺎﻓـﺖ دارو ﻧﺸـﺎن داد. ﺑﺎ ﺑﺮرﺳﯽ اﯾﻦ ﺗﻐﯿﯿﺮات و ﺷﻨﺎﺳﺎﯾﯽ ﺟﺎﯾﮕﺎه ﻣﺘﺎﺑﻮﻟﯿﺖ ﻫﺎي ﻣﺬﮐﻮر در ﻣﺴﯿﺮﻫﺎي ﻣﺘﺎﺑﻮﻟﯿﮑﯽ ﺗﻐﯿﯿﺮ ﯾﺎﻓﺘﻪ ﻣﯽ ﺗﻮان ﺟﻬـﺖ از ﺑـﯿﻦ ﺑﺮدن اﻧﮕﻞ در ﺟﺴﺘﺠﻮي ﻫﺪف دار ژن ﻫﺎي ﻣﻮﺛﺮ در ﺑﯿﺎن آﻧﺰﯾﻤﯽ اﯾﻦ ﻣﺴﯿﺮﻫﺎ ﺑـﻮد ﮐـﻪ ﺑـﺮا ي اﻧﮕـﻞ ﺿـﺮور ي ﻫﺴـﺘﻨﺪ و ﮐﻨﺘـﺮل ﻣﺘﺎﺑﻮﻟﯿﮑﯽ ﺑﺎﻻﯾﯽ را در اﻧﮕﻞ اﻧﺠﺎم داده در ﺣﺎﻟﯽ ﮐﻪ ﮐﻨﺘﺮل ﮐﻤﯽ در ﻣﯿﺰﺑﺎن دارﻧﺪ. ﺑﺎ ﺗﻮﺟﻪ ﺑﻪ اﻫﻤﯿﺖ درﻣﺎن ﺑﯿﻤـﺎر ي ﻟﯿﺸـﻤﺎﻧﯿﻮز و ﻧﺘﺎﯾﺞ ﻣﻄﺎﻟﻌﻪ ﺣﺎﺿﺮ از ﻣﺴﯿﺮﻫﺎي ﻣﺘﺎﺑﻮﻟﯿﺴﻤﯽ ﺗﻐﯿﯿﺮ ﯾﺎﻓﺘﻪ در اﻧﮕﻞ ﻟﯿﺸﻤﺎﻧﯿﺎ، ﺑﺎ ﺗﻤﺮﮐﺰ ﺑـﯿﺶ ﺗـﺮ روي اﯾﻦ ﻣﺴـﯿﺮﻫﺎ راﻫـي ﺑـﺮا ي ﺑﺮرﺳﯽ ﻫﺎي آﺗﯽ ﺑﯿﻤﺎري و ﮐﺸﻒ داروﻫﺎ و درﻣﺎن ﻫﺎي ﺟﺪﯾﺪ ﺑﺎز ﻣﯽ ﮔﺮدد.

Introduction: Based on WHO classifications, leishmaniasis enumerates as one of the six important diseases in the tropics. It is also common in the Middle East and the Mediterranean countries. Control of the disease is important due to increased clinical cases in immune-system deficiency patients. With regard to the increasing need to new drugs with lesser side effects for the control and treatment of the disease, we surveyed the in vitro effects of artemisinin on the promastigotes of leishmania major, s metabolites in Pasteur institute of Iran. Materials and Methods: After IC50 determination, the effects of different concentrations of artemisinin drug were assessed on the metabolites of leishmania in two groups ; the control group (without drug reception in culture media) and the case group (with drug reception in culture media), by using NMR technique. Results: The results of this study indicate that in the metabolic changes in promastigotes of leishmania major (fredlin strain) at stationary phase, 5 metabolites of the 41 ones in the galactose metabolic cycle have shown significant changes. Therefore, galactose metabolism has changed the most, while sphingolipid, valine, leucine and isoleucine biosynthesis follow respectively. Conclusion: Artemisinin resulted in metabolic changes in biochemical pathways including galactose, sphingolipids and also valine, lucine, isoleucine that culminated in cessation of leishmania major’s (fredlin strain) activity. Spectra derived from H NMR showed significant difference (p<0.05) between metabolites of the drug received group and the non-drug received group. The identification of altered metabolic sites in these pathways could be used for the annihilation of the parasite. Thus, detecting the important genes for the parasite in these pathways, with high metabolic control over the parasite, but minimal control over the host, is needed. Considering the importance of leishmaniasis treatment and the present data about modified metabolism pathways, we could present a novel route for the future assessment of the disease, drug discovery and new treatment with more focus on these pathways.