OBJECTIVE To investigate the secondary metabolite flavonoids containing beside epigallo catechin 3-gallate(EGCG)from n-hexane,chloroform and ethanolic fraction,cytotoxic potential from some fractions of Scurrula artro...OBJECTIVE To investigate the secondary metabolite flavonoids containing beside epigallo catechin 3-gallate(EGCG)from n-hexane,chloroform and ethanolic fraction,cytotoxic potential from some fractions of Scurrula artropurpurea(Blume)Danser(SAD),and its potential as DNA methyl transferase(DNMT)inhibitor.METHODS To identify 3 isolate from different regions,Lawang and Lembang(plateau region),Wlingi(lowland region)by 3fractions(n-hexane,chloroform and ethanol)using thin layer chromatography(TLC)and a spectrometer.EGCG level measured by Liquid-Chromatography mass spectrophotometer(LC-MS).The viability of HeLa cells measured by MTT assay.Acridine ethidium-bromide staining was applied to evaluate cell death.Molecular docking conducted for DNMT inhibition by Autodock Vina using Pyrex0.8 programs and molecular interaction by ligand scout V 2.0 program.RESULTS The different region showed different secondary metabolite.SAD from Lawang and Lembang(both are polite)have similar active compound in n-hexan fraction(didhydroflavonol,flavonone,flavanone),chloroform fraction(dihydroflavonol,flavanone,catechin)and ethanolic fraction(flavonone,flavonol and EGCG).While Wlingi isolate have no dihydroflavonol in chloroform fraction,but flavone.We used Lawang isolate for next research to evaluate cytotoxicity potential for HeLa cells for 24 h.IC50chloroform fraction was the highest,followed by ethanol and nhexane fraction(96.16,298.82 and 489.66μg·mL-1,respectively).Index apoptosis were observed at half IC50 concentration of chloroform(99.97%),ethanol(66.79%)and n-hexane fractions(14.92%).In silico analysis showed some active compounds have specific binding site to the DNMT with low energy binding affinity such as-10.4for EGCG,-8.4for catechin,-8.2 for dihydroflavonol,-8.0 for flavone,-7.9 for flavonol and -7.8kcal·mol-1 for flavonone.The impact of of DNMT and bioactive compounds complex showed its potential for epigenetic manipulation such as demethylation.Inhibition of DNMT on ABCG2 gene could combat drug efflux which main problem of multidrug resistance in cervical cancer chemotherapy.CONCLUSION Chloroform fraction(without EGCG containing)have the highest potential for cytotoxic agent for Hela cells rather than ethanol and n-hexane fraction and could develop as chemotherapeutic agent due to its property against multi drugs resistance as well.展开更多
基金The project supported by Research Development Unit of Medical Faculty,Brawijaya University 2014
文摘OBJECTIVE To investigate the secondary metabolite flavonoids containing beside epigallo catechin 3-gallate(EGCG)from n-hexane,chloroform and ethanolic fraction,cytotoxic potential from some fractions of Scurrula artropurpurea(Blume)Danser(SAD),and its potential as DNA methyl transferase(DNMT)inhibitor.METHODS To identify 3 isolate from different regions,Lawang and Lembang(plateau region),Wlingi(lowland region)by 3fractions(n-hexane,chloroform and ethanol)using thin layer chromatography(TLC)and a spectrometer.EGCG level measured by Liquid-Chromatography mass spectrophotometer(LC-MS).The viability of HeLa cells measured by MTT assay.Acridine ethidium-bromide staining was applied to evaluate cell death.Molecular docking conducted for DNMT inhibition by Autodock Vina using Pyrex0.8 programs and molecular interaction by ligand scout V 2.0 program.RESULTS The different region showed different secondary metabolite.SAD from Lawang and Lembang(both are polite)have similar active compound in n-hexan fraction(didhydroflavonol,flavonone,flavanone),chloroform fraction(dihydroflavonol,flavanone,catechin)and ethanolic fraction(flavonone,flavonol and EGCG).While Wlingi isolate have no dihydroflavonol in chloroform fraction,but flavone.We used Lawang isolate for next research to evaluate cytotoxicity potential for HeLa cells for 24 h.IC50chloroform fraction was the highest,followed by ethanol and nhexane fraction(96.16,298.82 and 489.66μg·mL-1,respectively).Index apoptosis were observed at half IC50 concentration of chloroform(99.97%),ethanol(66.79%)and n-hexane fractions(14.92%).In silico analysis showed some active compounds have specific binding site to the DNMT with low energy binding affinity such as-10.4for EGCG,-8.4for catechin,-8.2 for dihydroflavonol,-8.0 for flavone,-7.9 for flavonol and -7.8kcal·mol-1 for flavonone.The impact of of DNMT and bioactive compounds complex showed its potential for epigenetic manipulation such as demethylation.Inhibition of DNMT on ABCG2 gene could combat drug efflux which main problem of multidrug resistance in cervical cancer chemotherapy.CONCLUSION Chloroform fraction(without EGCG containing)have the highest potential for cytotoxic agent for Hela cells rather than ethanol and n-hexane fraction and could develop as chemotherapeutic agent due to its property against multi drugs resistance as well.
