اﺛﺮ ﻋﺼﺎره ﻫﯿﺪرواﺗﺎﻧﻠﯽ داﻧﻪ ﺧﺮﺑﺰه Cucumis melo L ﺑﺮ ﺑﯿﺎن ﮐﺒﺪي ژن ﻫﺎي ﮐﺪ ﮐﻨﻨﺪه NADPH اﮐﺴﯿﺪاز و ﮔﻠﻮﺗﺎﺗﯿﻮن ﭘﺮاﮐﺴﯿﺪاز در ﻣﻮش ﻫﺎي ﺻﺤﺮاﯾﯽ ﻣﺴﻤﻮم ﺷﺪه ﺗﻮﺳﻂ اﺗﯿﻠﻦ ﮔﻠﯿﮑﻮل

Effect of hydro-ethanolic extract of Cucumis melo seeds on hepatic expression of genes encoding NADPH-oxidase and glutathione peroxidase in ethylene glycol-induced toxicity in male rats

ﻣﺴﻤﻮﻣﯿﺖ ﮐﺒﺪي ﻫﻤﯿﺸﻪ ﺑﺎ اﺳﺘﺮس اﮐﺴﯿﺪاﺗﯿﻮ و اﻟﺘﻬﺎب ﻫﻤﺮاه اﺳﺖ و آﻧﺘﯽاﮐﺴﯿﺪان ﻫﺎ ﻣﻮﺟﺐ ﮐﺎﻫﺶ اﺛﺮات ﻋﻮاﻣﻞ ﺳﻤﯽ ﺑﺮ ﮐﺒﺪ ﻣﯽ ﺷﻮﻧﺪ. ﻓﻌﺎﻟﯿﺖ آﻧﺘﯽ اﮐﺴﯿﺪاﻧﯽ ﮔﯿﺎﻫﺎن ﻣﮑﺎﻧﯿﺴﻢ ﮐﻠﯽ اﺳﺖ ﮐﻪ ﺗﻮﺳﻂ آن اﮐﺜﺮ ﮔﯿﺎﻫﺎن از ﮐﺒﺪ ﻣﺤﺎﻓﻈﺖ ﻣﯽ ﮐﻨﻨﺪ. در ﻣﻄﺎﻟﻌﻪ ﺣﺎﺿﺮ ﺗﺎﺛﯿﺮ ﻋﺼﺎره ﻫﯿﺪرواﺗﺎﻧﻠﯽ داﻧﻪ ﺧﺮﺑﺰه ﺑﺮ ﺳﻄﺢ ﺑﯿﺎن ژن ﻫﺎي Nox4 و gpx1 در ﻣﻮش ﻫﺎي ﻣﺴﻤﻮم ﺷﺪه ﺑﺎ اﺗﯿﻠﻦ ﮔﻠﯿﮑﻮل ﻣﻮرد ارزﯾﺎﺑﯽ ﻗﺮار ﮔﺮﻓﺖ. در ﻣﻄﺎﻟﻌﻪ ﺣﺎﺿﺮ در اﺑﺘﺪا ﻋﺼﺎره داﻧﻪ ﺧﺮﺑﺰه ﺑﻪ روش ﺧﯿﺴﺎﻧﺪن در اﺗﺎﻧﻞ 80 درﺻﺪ ﺗﻬﯿﻪ ﺷﺪ، ﻣﻮش ﻫﺎ در 5 ﮔﺮوه 6 ﺗﺎﯾﯽ ﺑﻪﺻﻮرت ﯾﮏ ﮔﺮوه ﮐﻨﺘﺮل ﺳﺎﻟﻢ، ﯾﮏ ﮔﺮوه ﮐﻨﺘﺮل ﻣﺴﻤﻮم و ﺳﻪ ﮔﺮوه ﻣﺴﻤﻮم ﺗﺠﺮﺑﯽ ﺗﻘﺴﯿﻢ ﺷﺪﻧﺪ. ﺑﻪ ﮔﺮوه ﻣﺴﻤﻮم ﺗﺠﺮﺑﯽ ﻏﻠﻈﺖ ﻫﺎي 300 ،150 و 600 ﻣﯿﻠﯽﮔﺮم ﺑﺮ ﮐﯿﻠﻮﮔﺮم ﻋﺼﺎره ﻫﯿﺪرواﺗﺎﻧﻠﯽ داﻧﻪ ﺧﺮﺑﺰه ﺑﻪﺻﻮرت ﺧﻮراﮐﯽ ﺧﻮراﻧﺪه ﺷﺪ. ﻣﻮشﻫﺎ ﺑﻪﻣﺪت 38 روز ﺗﯿﻤﺎر ﺷﺪه و ﺳﭙﺲ ﮐﺒﺪ ﻣﻮش ﻫﺎ ﺧﺎرج ﺷﺪه و ﺳﭙﺲ RNA ﮐﻞ اﺳﺘﺨﺮاج ﺷﺪ. ﺳﭙﺲ cDNA ﺳﺎﺧﺘﻪ ﺷﺪه و ﻣﯿﺰان ﺑﯿﺎن ژن ﻫﺎي ﻣﻮرد ﻧﻈﺮ و ﻫﻤﯿﻦ ﻃﻮر ژن GAPDH ﺑﻪﻋﻨﻮان ژن ﺧﺎﻧﻪ دار ﺑﺎ روش Real time PCR ﻣﻮرد ارزﯾﺎﺑﯽ ﻗﺮار ﮔﺮﻓﺖ. ﻧﺘﺎﯾﺞ ﻣﻄﺎﻟﻌﻪ ﺣﺎﺿﺮ ﻧﺸﺎن ﻣﯽ دﻫﺪ ﺗﯿﻤﺎر روزاﻧﻪ ﺧﻮراﮐﯽ اﺗﯿﻠﻦ ﮔﻠﯿﮑﻮل ﻣﻮﺟﺐ اﻓﺰاﯾﺶ ﻣﻌﻨﯽ دار ﺑﯿﺎن ژن ﮐﺪﮐﻨﻨﺪه nox4 و gpx1 در ﮔﺮوه ﻫﺎي ﻣﺴﻤﻮم ﻧﺴﺒﺖ ﺑﻪ ﮐﻨﺘﺮل ﺳﺎﻟﻢ ﻣﯽ ﺷﻮد. اﻓﺰاﯾﺶ ﺑﯿﺎن ژن nox4 و gpx1 ﻧﺸﺎن دﻫﻨﺪه وﻗﻮع اﻟﺘﻬﺎب ﻧﺎﺷﯽ از ﻣﺼﺮف اﺗﯿﻠﻦ ﮔﻠﯿﮑﻮل و ﻣﺴﻤﻮﻣﯿﺖ آن ﻣﯽ ﺑﺎﺷﺪ. ﺗﯿﻤﺎر ﻋﺼﺎره ﻫﯿﺪرواﺗﺎﻧﻠﯽ داﻧﻪ ﺧﺮﺑﺰه در ﻏﻠﻈﺖ ﻫﺎي 300 ،150 و 600 ﻣﯿﻠﯽ ﮔﺮم ﺑﺮ ﮐﯿﻠﻮﮔﺮم وزن ﺑﺪن ﻣﻮﺟﺐ ﮐﺎﻫﺶ ﻣﻌﻨﯽ دار ﺑﯿﺎن ﻫﺮ دو ژن در ﮔﺮوهﻫﺎي ﺗﺠﺮﺑﯽ در ﻣﻘﺎﯾﺴﻪ ﺑﺎ ﮔﺮوه ﮐﻨﺘﺮل ﻣﺴﻤﻮم ﻣﯽ ﺷﻮد. ﮐﺎﻫﺶ ﻣﯿﺰان ﺑﯿﺎن ژن nox4 در ﻣﻘﺎﯾﺴﻪ ﺑﺎ ﮔﺮوه ﮐﻨﺘﺮل ﻣﺴﻤﻮم ﻧﺸﺎن ﻣﯽ دﻫﺪ ﻣﺼﺮف ﻋﺼﺎره ﻫﯿﺪرواﺗﺎﻧﻠﯽ داﻧﻪ ﺧﺮﺑﺰه ﻣﻮﺟﺐ ﮐﺎﻫﺶ اﻟﺘﻬﺎب ﻧﺎﺷﯽ از ﻣﺴﻤﻮﻣﯿﺖ ﺑﺎ اﺗﯿﻠﻦ ﮔﻠﯿﮑﻮل ﺷﺪه اﺳﺖ. ﺑﺎ ﺗﻮﺟﻪ ﺑﻪ اﯾﻦﮐﻪ ﺑﻪدﻧﺒﺎل ﻣﺴﻤﻮﻣﯿﺖ ﮐﺒﺪي ﻧﺎﺷﯽ از اﺗﯿﻠﻦ ﮔﻠﯿﮑﻮل ﻣﯿﺰان H2O2 اﻓﺰاﯾﺶ ﻣﯽ ﯾﺎﺑﺪ ﺑﻪﻫﻤﯿﻦ دﻟﯿﻞ ﻣﯿﺰان ﺑﯿﺎن ژن ﮐﺪﮐﻨﻨﺪه ﮔﻠﻮﺗﺎﺗﯿﻮن ﭘﺮاﮐﺴﯿﺪاز )gpx( اﻓﺰاﯾﺶ ﻣﯽ ﯾﺎﺑﺪ. ﺑﻪدﻧﺒﺎل ﺗﯿﻤﺎر ﺑﺎ ﻋﺼﺎره ﻫﯿﺪرواﺗﺎﻧﻠﯽ ﺧﺮﺑﺰه، ﻣﯿﺰان ﺗﻮﻟﯿﺪ H2O2 ﮐﺎﻫﺶ ﻣﯽ ﯾﺎﺑﺪ ﮐﻪ اﯾﻦ ﮐﺎﻫﺶ ﻣﯽ ﺗﻮاﻧﺪ ﺑﻪدﻟﯿﻞ اﺛﺮات ﺗﺮﮐﯿﺒﺎت آﻧﺘﯽ اﮐﺴﯿﺪاﻧﯽ ﻣﻮﺟﻮد در ﻋﺼﺎره ﺑﺎﺷﺪ. ﺑﻨﺎﺑﺮاﯾﻦ ﺑﻪدﻟﯿﻞ ﮐﺎﻫﺶ H2O2 ﻋﺎﻣﻞ ﻣﺤﺮك ﺑﺮ اﻓﺰاﯾﺶ ﺑﯿﺎن ژن ﮔﻠﻮﺗﺎﺗﯿﻮن ﭘﺮاﮐﺴﯿﺪاز )gpx( وﺟﻮد ﻧﺪاﺷﺘﻪ و ﺑﻨﺎﺑﺮاﯾﻦ ﻣﯿﺰان ﺑﯿﺎن اﯾﻦ ژن ﻧﯿﺰ ﮐﺎﻫﺶ ﻣﯽ ﯾﺎﺑﺪ. ﺑﻨﺎﺑﺮاﯾﻦ، ﺑﺎ ﺗﻮﺟﻪ ﺑﻪ ﮐﺎﻫﺶ ﻣﯿﺰان ﺑﯿﺎن ﻫﺮ دو ژن ﮔﻠﻮﺗﺎﺗﯿﻮن ﭘﺮاﮐﺴﯿﺪاز )gpx( و NADPH اﮐﺴﯿﺪاز )nox4( و ارﺗﺒﺎط ﻣﯿﺰان ﺑﯿﺎن ﻫﺮ دو ژن ﺑﺎ ﻣﯿﺰان H2O2، ﺑﻪﻧﻈﺮ ﻣﯽ رﺳﺪ ﻋﺼﺎره ﻫﯿﺪرواﻟﮑﻠﯽ داﻧﻪ ﺧﺮﺑﺰه ﺑﻪدﻟﯿﻞ اﺛﺮات آﻧﺘﯽ ﮐﺴﯿﺪاﻧﺘﯽ ﻣﻮﺛﺮ ﻣﻮﺟﻮد در آن ﻣﯿﺰان H2O2 ﮐﺎﻫﺶ ﯾﺎﻓﺘﻪ و ﺑﻪﻫﻤﯿﻦ دﻟﯿﻞ ﻣﯿﺰان ﺑﯿﺎن اﯾﻦ دو ژن ﻧﯿﺰ ﮐﺎﻫﺶ ﻣﯽ ﯾﺎﺑﺪ.

effects of toxins on the liver. The antioxidant activity of plants is the general mechanism by which most plants protect the liver. In the present study, the effect of hydroanthanolic extract of melon seed on the expression level of NOX4 and GPx1 genes in ethylene glycol poisoned mice was evaluated. In the present study, melon seed extract was first prepared by soaking in 80% ethanol. Mice were divided into 5 groups in each group 6 rat: a healthy control group, a poisoned control group and three experimental poisoned groups. To the experimental poisoned group, concentrations of 150, 300 and 600 mg / kg of hydroethanolic extract of melon seed were fed orally. The mice were treated for 38 days and then the mice's liver was removed and then total RNA was extracted. Then cDNA was constructed and the expression of the desired genes as well as GAPDH gene as housewife gene was evaluated by Real time PCR. The results of the present study show that daily oral administration of ethylene glycol significantly increases the expression of genes encoding NOX4 and GPx1 in toxic groups compared to healthy controls. Increased expression of NOX4 and GPx1 genes indicates the occurrence of ethylene glycol-induced inflammation and toxicity. Treatment with hydroethanolic extract of melon seed at concentrations of 150, 300 and 600 mg / kg body weight significantly reduced the expression of NOX4 and GPx1 genes in the experimental groups compared with the control group. Decreased expression of NOX4 gene in comparison with the poisoned control group shows that administration of hydro-ethanolic extract of melon seed has reduced inflammation caused by ethylene glycol toxicity. Due to the fact that the amount of H2O2 increases following hepatotoxicity caused by ethylene glycol, the expression of the gene encoding glutathione peroxidase (GPx) increases. Following treatment with melon hydro-ethanolic extract, H2O2 production is reduced, which may be due to the effects of antioxidant compounds in the extract. Therefore, due to the decrease in H2O2, there is no stimulus for increasing the expression of glutathione peroxidase (GPx) gene, and therefore the expression of this gene also decreases. In addition, due to the decrease in the expression of both glutathione peroxidase (GPx) and NADPH oxidase (NOx4) genes and the relationship between the expression of both genes and H2O2, it seems that the hydro-ethanolic extract of melon seed decreased H2O2 due to its effective antioxidant effects and therefore the expression of these genes are reduced.