| Mangosteen(Garcinia mangostana L.)is a fruit widely cultivated in the tropics.It is rich in a variety of nutrients and has extremely high medicinal value.In recent years,there have also been some reports on the extract of mangosteen and its active components.Malaria is transmitted through mosquito bites or blood transfusions infected with malaria parasites.Inhibition of the activity of quinone reductase 2(QR-2,NQO2)in red blood cells leads to the accumulation of superoxide radicals in cells,which may be responsible for killing the parasites.Protein tyrosine phosphatase 1 B(PTP1B)is a kind of important regulation of glucose homeostasis and energy metabolism,insulin and leptin signaling pathway plays an important role in the negative control,excessive PTP1 B expression,reducing the activity of protein tyrosine kinase,thereby weakening insulin receptor binding of insulin,insulin resistance,eventually leading to type 2 diabetes.In this study,UF-HPLC-QTOF/MS was used to screen and identify possible QR-2 and PTP1 B inhibitors in the blood components of mangosteen extract.Components extracted by heat ethanol refluxing extraction,blood samples were collected from rats following the oral administration of mangosteen extract and then incubated with QR-2 followed by UF-HPLC-QTOF/MS analysis to rapidly screen for and identify the QR-2-inhibiting xanthones.A total of 16 kinds of xanthone compounds were identified,which were mangostenone C,1,3,6,7-tetrahydroxy-8-prenylxanthone,Garcinone C,Garcimangosone C,1,5,8-trihydroxy-3-methoxy-2-(3-methylbut-2-enyl)xanthone,Garcinone D,mangostanol,1,7 dihydroxy-2-(3-methylbut-2-enyl)-3-methoxyxanthone gamma mangostin,8-deoxygartanin,1,3,7-trihydroxy-2,8-di-(3-methylbut-2-enyl)-xanthone,alpha mangostin,garcinone E,9-hydroxycalabaxanthone,beta mangostin,tovophyllin A.Then,six compounds(γ-mangostin,8-deoxyartanin,1,3,7-trihydroxy-2,8-di-(3-methylbut-2-enyl)-xanthone,α-mangostin and 9-hydroxycalabaxanthone)were selected for in vitro enzymatic activity assay.The results showed that γ-mangostin exhibited the strongest inhibitory activity with IC50 value of 3.82±0.51 μM.Docking experiments with QR-2 indicated the relationship between the structure and the inhibitory activity.There was the least residual interaction with QR-2,and the inhibition activity was the least,which might be due to the fact that there were fewer phenolic hydroxyl groups and isoprene groups in the structure of xanthone.In addition,the results of pharmacokinetics showed that all the six oxanthrones could be distributed rapidly in the body with a second peak phenomenon,and the hepatointestinal circulation might be the main reason for the double peak phenomenon of the analytes.The mangosteen fruit and its main active ingredient(xanthones)can inhibit the growth of Plasmodium falciparum,but the mechanism of action is still unclear.Therefore,we hypothesized that the presence of some ingested mangosteen components in the blood exerted an antimalarial effect by inhibiting QR-2 activity,which may serve as the theoretical basis for on-demand mangosteen consumption to help prevent malaria.In this study,the UF-HPLCQTOF/MS method was used to screen the blood samples of rats after oral administration of mangosteen extract to determine whether compounds in mangosteen can inhibit QR-2 activity.In addition,the pharmacokinetic analysis of mangosteen extract orally in rats was carried out and the significance of the experiment was discussed.The results of this study may provide some theoretical support for the benefits of regular mangosteen consumption in preventing malaria.Components extracted by heat ethanol refluxing extraction,extracts of animals to fill the stomach,and to take blood,eyes take blood method for serum protein deposition process,get extract serum after incubation with PTP1 B,UF-HPLC-QTOF/MS method for components of extracts in blood of rats after the treatment with PTP1 B complexes formed by the compound screening,Six xanthones(γ-mangostin,8-deoxyartanin,1,3,7-trihydroxy-2,8-di-(3-methylbut-2-enyl)-xanthone,α-mangostin,garcinone E and 9-hydroxycalabaxanthone)were identified from the blood of mice and the enzyme activity inhibition test was carried out in vitro.Garcinone E was the most effective PTP1 B inhibitor with IC50 value of 0.43±0.11μM.The tissue distribution test by UHPLC-QQQ method showed that these xanthones were absorbed rapidly and distributed widely in different tissues.The experiment results show that found that part of the components of extracts xanthone can also improve the symptoms of diabetes,the components of these xanthone can produce inhibition of PTP1 B after being absorbed.In addition,the findings of this study may also provide some clues as to the daily intake of mangosteen needed to mitigate the effects of diabetes.In order to make full use of mangosteen resources,the extraction rate of six main active components(γ-mangostin,8-deoxyartanin,1,3,7-trihydroxy-2,8-di-(3-methylbut-2-enyl)-xanthone,α-mangostin and 9-hydroxycalaba)in mangosteen extract was increased.Firstly,the single factor of heating extraction method,ultrasoundassisted heating extraction method and ball mill extraction method were investigated.The effects of liquid-solid ratio,extraction time and ethanol concentration on the extraction yield were investigated respectively.The response surface analysis(RSM)was used to optimize the extraction process to determine the best extraction process.Taking γ-mangostin as an example,the actual extraction parameters of the heating extraction method were as follows: extraction time 150 min,extraction temperature70 ℃,liquid-solid ratio 40 m L/g,the actual yield was 0.711%,and the relative error was about 2.1% compared with the theoretical predicted value.The actual extraction parameters of the ball mill extraction method were as follows: extraction time 30.0 min,power of the ball mill 20.0 Hz,liquid/solid ratio 32 m L/g.The actual measured yield was 0.534%,and the relative error was about 4.5% compared with the theoretical predicted value.The ultrasonic assisted heating extraction method was not suitable for the extraction of six oxanthrones in mangosteen due to its long extraction time.In conclusion,this study can provide a new perspective for the potential application of mangosteen as a functional food ingredient in inhibiting QR-2 or PTP1 B activity.However,the required daily intake of mangosteen and the exact contribution of mangosteen to malaria control remain to be studied.In addition,the findings of this study may also provide some clues as to the daily intake of mangosteen required to mitigate the effects of diabetes with inhibitory activity.The exact benefits of mangosteen for diabetes,as well as the extent and time required for ingesting mangosteen to have a positive effect,are still subject to further research. |
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