Aktivitas Antioksidan dan Penghambat Xantin Oksidase dari Ekstrak Buah Andaliman (Zanthoxylum acanthopodium DC.)

The purpose of this study was to evaluate the antioxidant activities, tyrosinase inhibitory effects on the fruiting bodies of Pleurotus salmoneostramineus extracted with acetone, methanol and hot water. The antioxidant activities were performed on β-carotene-linoleic acid, reducing power, DPPH, ferrous ions chelating abilities, and xanthine oxidase inhibitory activities. In addition to this, phenolic compounds were also detected. Acetonic, methanolic and hot water extracts of P. salmoneostramineus showed the similar pattern of β-carotene-linoleic acid inhibition. At 8 mg/ml, methanolic extract showed a high reducing power of 1.62. The scavenging effects on DPPH, acetonic and methanolic extracts were effective than hot water extract. The strongest chelating effect was obtained from the acetonic and methanolic extracts at 1.0 mg/ml. Gallic acid, protocatechuic acid, chlorogenic acid, formononetin, and biochanin-A were detected from acetonitrile and 0.1N hydrochloric acid (5:1) solvent extract. The xanthine oxidase and tyrosinase inhibitory activities of the acetonic, methanolic, and hot water extracts increased with increasing concentration. The results suggested that fruiting bodies of P. salmoneostramineus may have potential as a natural antioxidants. Key words: Antioxidant, phenolic compounds, Pleurotus salmoneostramineus, tyrosinase inhibition, xanthine oxidase.

Euphorbia heterophylla Linn (Euphorbiaceae) is a medicinal plant used by traditional herbal practitioners in Nigeria and some parts of West Africa for the treatment of constipation, bacterial and inflammatory disease conditions (arthritis and rheumatism). Powdered plant material was subjected to phytochemical screening using standard experimental procedures. The crude powdered sample of Euphorbia heterophylla leaves was extracted with n-hexane, chloroform, ethylacetate and methanol. The precipitates from the fractions were subjected to chromatographic and re-crystallization procedures. The structures of isolated compounds were characterized and elucidated with chemical and spectroscopic techniques such as IR, NMR and MS experiments. The in vitro biological activity of the isolated and characterized compounds were evaluated by superoxide scavenging assay using xanthine – xanthine oxidase system to generate superoxides. The result of the study showed that the crude plant material contained some secondary metabolites such as saponins, flavonoids and tannins. The phytochemical investigation led to the isolation of four known compounds stigmasterol, β-stigmasterol glucoside, benzoic acid and 4 – hydroxyl benzoic acid. The four compounds exhibited good activity against the xanthine oxidase enzymes while the 4-hydroxybenzoic acid showed a marked activity. The isolation of the compounds from the leaves of E. heterophylla, which inhibited the xanthine oxidase enzyme, has justified the claims for which the plant is known and used. Key words: Euphorbia heterophylla, euphorbiaceae, xanthine oxidase activity, superoxide scavenging, steriods.

Serum uric acid was be elevated because of urate overproduction and/or underexcretion, and it will cause hyperuricemia. Hyperuricemia may cause the deposition of uric acid in joints and lead to painful inflammation. There were also many researches indicated that hyperuricemia was related with atherosclerosis, hypertension, cardiovascular diseases, diabetes mellitus and renal disease. Xanthine oxidase (XO) will catalyze the hypoxanthine and xanthine to uric acid, which is the main enzyme in hyperuricemia. The current agent used to treat the hyperuricemia is XO inhibitor, but the allopurinol used in clinical will cause side effect and prompt the development of XO inhibitor. Reactive oxygen species (ROS) had been known to cause lipid peroxidation which induce nutritional degradation of food products. ROS were also related to chronic cardiac failure, ageing, acute respiratory distress syndrome. Because of the side effect of the medicine and the fashion of healthy food, many scholars start to pay attention to the research of non side effect or low side effect compound from traditional medicine. Among these researches, most scholars are focusing on XO inhibitor agents and antioxidant. Although the natural antioxidant is safe in general, daily usage may cause potential complication, for example it may affect the function of liver and kidney or it may also impact on fertilized egg implantation and fetal organogenesis when the rats were oral administration with high dosage during the period of pregnancy. Therefore this study was investigated to the bioactivity and toxicity of extractive from the leaves of Tradescantia albiflora Kunth, which is a Chinese medicine plant to reduce serum uric acid level on rat. The experiment was designed to use oxonate to be injected intraperitonelly to induce rat in hyperuricemic model, then these rats were oral administration crude extractive from the leaves, stem of T. albiflora or PBS respectively. Blood samples were collected to process hematology and biochemistry study every 90 mins, and monitored the change of blood pressure in the same time. We have used hexane, ethyl acetate, butanol and distilled water to extract the dry leaves of T. albiflora then got hexane extractive (HE), ethyl acetate extractive (EAE), butanol extractive (BE) and water extractive (WE) respectively. The experiment of decrease the plasma uric acid (PUA) level with extractives used by rat in hyperuricemic model to oral administration with HE, EAE, BE and WE extractive respectively, and bloods samples were collected to process PUA test every 30 mins. The experiments of eliminated uric acid were utilized by rat in hyperuricemic model to oral administration with the PBS or WE respectively, and urine samples were collected to process urine uric acid (UUA) test after 30、90、150 and 210 mins. These tests of the efficiency for scavenge of DPPH and the ferric thiocyanate (FTC) have utilized different concentration of extractives belonging to 4 layers react with agents, and tested the absorption with 517 and 500 nm respectively. Crude extractives were divided into three groups. One was the control group (oral phosphate buffer). Second group was tested for oral consumption of 50 times the effective dosage of reducing plasma uric acid level (625 mg/kg) for 1~4 days during pregnancy. Third group was tested during 8~11 days of pregnancy. Subacute toxicities were tested the consumption of the 10 times effective dosage of reducing plasma uric acid level (125 mg/kg) for 28 days and blood samples were collected to be conducted hematology, biochemistry and histology study. The results of these studies indicated that T. albiflora can decrease the level of blood uric acid, these 4 different types of extractive had the same ability, but the WE had the best affect in last estimative point (240 min). These 4 different types of extractive can also inhibit the XO, and WE had the best efficacy. The result of eliminated uric acid test revealed that WE hadn’t affect to eliminate uric acid. The BE had better efficacy in the scavenge of DPPH testing and the FTC testing. The toxicological result showed that fertilized egg implantation rate, fetal organogenesis and resorption rate were not be impacted on the femate rat after oral administrateion with 50 times the effective dosage extractives of reducing plasma uric acid level (625 mg/kg) from the leaves of T. albiflora extractive during the pregnant period.The test of subacute toxicity revealed the Hb, HCT and RBC had significant increase (p＜0.05); the ALKP and BUN had significant decrease (p＜0.05) compared with control group. This study has shown that T. albiflora can decrease the blood uric acid by inhibiting XO, and it also can eliminate free radical by preventing fatty oxidantion to achieve antioxidant. Female rat oral administrated high dosage during the period of pregnancy may not impact fertilized egg implantation, fetal organogenesis and resorption, but oral administrated T. albiflora for long and high dosage may cause hepatic cell swelling and slightly hyperplasia of bile duct, it could suggest that T. albiflora is not suitable to medicate in long and high dosage.

Peperomia pellucida (L.) Kunth known as Suruhan is a potential medicinal plants, used traditionally to treat gout. Suruhan herb (Peperomia pellucida (L.) Kunth) had been studied in vivo, and found to be able to lowering uric acid levels in the blood. Ethanol extract of Peperomia pellucida (L.) Kunth herb also had been studied in vitro, and found to be potential to inhibit xanthine oxidase. The purpose of this study is to know the potential of fraction from ethanol extract of Peperomia pellucida (L.) Kunth. herb as xanthine oxidase inhibitor compared to allopurinol and its ethanolic extract. Ethanol extract of Peperomia pellucida (L.) Kunth herb was obtained by percolation method using ethanol 96%. The fractionation is done by column chromatography method using silica as stationary phase and n-hexane, n-hexane-ethyl acetate, ethyl acetate, ethyl acetate-ethanol, and ethanol as mobile phases. The fraction which contained flavonoid compounds was tested its xanthine oxidase inhibition potency using UV spectrophotometer at λ 290 nm. The absorbance was observed every 10 seconds for 10 minutes for extract and fraction with the concentration of 0.25 ppm - 5 ppm, while allopurinol was determined with the concentration of 0.2 ppm - 3.2 ppm. The result showed that ethyl acetate-ethanol fraction potentially inhibited xanthine oxidase with IC50 value of 5.00 ± 0.06 ppm, while ethanolic extract of Peperomia pellucida (L.) Kunth herb and allopurinol have IC50 value 0.33± 0.07 ppm and 0.84 ± 0.02 ppm respectively. Thus it can be concluded that ethyl acetateethanol fraction had potential as xanthine oxidase inhibitor, but the potential is lower than the ethanolic extract of Peperomia pellucida (L.) Kunth herb.

Objective: The study deals with the evaluation of the in vitro antimalarial activity of three crude extracts (ethanol and 80% methanol) from the leaves of Morinda morindoides (Baker) Milne-Redh. (Rubiaceae), that of their respective soluble fractions and isolated compounds, and that of the dichloromethane extract. It also reports the in vivo antimalarial activity of the three crude extracts and the petroleum ether soluble fraction from the ethanol extract as well as the toxicity of crude extracts in mice. Materials and methods: The ethanol, 80% methanol and dichloromethane extracts were obtained by maceration and percolation of powdered dried M. morindoides leaves. The ethanol extract was dissolved in 100 ml distilled water and extracted with petroleum ether, and then acidified with HCl 2N (pH 2-3) and successively and exhaustively treated with chloroform and isoamylic alcohol. On the other hand the 80% methanol extract was dissolved in 100 ml distilled water and successively and exhaustively extracted with chloroform, ethyl acetate and n-butanol. A series of flavonoids, anthraquinones and iridoids were isolated from the 80% methanol extract. All dried samples were tested for their potential in vitro antiplasmodial activity against Congolese the chloroquine-sensitive or the chloroquine-sensitive NF54/64, clone A19 strains of Plasmodium falciparum according to the case. The three extracts and one soluble fraction were tested in vivo against Plasmodium berghei berghei in a classical 4-suppressive test. Results: The petroleum ether, isoamylic alcohol and chloroform soluble fractions from the partition of ethanol extract showed an in vitro antiplasmodial activity against Congolese the chloroquine-sensitive strain of P. falciparum with IC 50 values of 1.8 ± 0.2, 15.3 ± 3.6 and 8.8 ± 2.5 μg/ml respectively. Only the chloroform soluble fraction from the partition of the 80% methanol exhibited good antiplasmodial activity with IC 50 value of 8.3 ± 1.6 μg/ml against the chloroquinesensitive NF54/64, clone A1A9 strain of P. falciparum . Among isolated compounds, quercetin exhibited good antiplasmodial activity with IC 50 value of 5.5. ± 1.8 μg/ml, alizarin and chrysarin showed a moderate activity with IC 50 ranging from 14 to 26 μg/ml. In vivo test, at a daily oral dose of 200 mg/kg, ethanol, 80% methanol and dichloromethane extracts, and the petroleum ether soluble fraction produced 33%, 54% and 73%, and 75% chemosuppression respectively. Conclusion: These results may partly justify and support the traditional use of Morinda morinoides leaves for the treatment of malaria.

The corneas of albino rabbits were irradiated (5 min exposure once a day) with UVB rays (312 nm) for 4 days (shorter procedure) or 8 days (longer procedure). The eyes were examined microbiologically and only the corneas of sterile eyes or eyes with non-pathogenic microbes were employed. Histochemically, the activities of reactive oxygen species (ROS)-generating oxidases (xanthine oxidase, D-amino acid oxidase and alpha-hydroxy acid oxidase) were examined in cryostat sections of the whole corneas. Biochemically, the activity of xanthine oxidoreductase/xanthine oxidase was investigated in the scraped corneal epithelium. UVB rays significantly changed enzyme activities in the corneas. In comparison to the normal cornea, where of ROS-generating oxidases only xanthine oxidase showed significant activity in the corneal epithelium and endothelium, D-amino acid oxidase was very low and alpha-hydroxy acid oxidase could not be detected at all, in the cornea repeatedly irradiated with UVB rays, increased activities of xanthine oxidase and D-amino acid oxidase were observed in all corneal layers. Only after the longer procedure the xanthine oxidase and D-amino acid oxidase activities were decreased in the thinned epithelium in parallel with its morphological disturbances. Further results show that the xanthine oxidase/xanthine oxidoreductase ratio increased in the epithelium together with the repeated irradiation with UVB rays. This might suggest that xanthine dehydrogenase is converted to xanthine oxidase. However, in comparison to the normal corneal epithelium, the total amount of xanthine oxidoredutase was decreased in the irradiated epithelium. It is presumed that xanthine oxidoreductase might be released extracellularly (into tears) or the enzyme molecules were denatured due to UVB rays (particulary after the longer procedure). Comparative histochemical and biochemical findings suggest that reactive oxygen species-generating oxidases (xanthine oxidase, D-amino acid oxidase) contribute to the corneal damage evoked by UVB rays.

Solanum nigrum Linn (SNL), a member of the Solanaceae family , is a common herb that grows wildly and abundantly in open fields .Traditionally, it has been used in chinese medicine for curing inflammatory disease and various kinds of cancers. Approximately, 25 % of all cancers are somehow associated with chronic infection and inflammation. Chronic inflammation is considered as a possible mechanism in initiating tumorigenesis, resulting from the generation of ROS (Reactive Oxygen Species)/RNS (Reactive Nitrogen Species) by macrophages in the inflamed tissue and resulted in DNA damage. Chronic dysregulated activation of some cellular ROS/Eicosanoid generating enzymes, NOX-2 (NADPH Oxidase-2), XO (Xanthine Oxidase),iNOS (inducible Nitric Oxide Synthase) and COX-2 (Cyclooxygenase-2), have been shown to play pivotal roles during this process. The signal transduction pathways related to these enzymes in macrophages mainly including NF-κB/AP-1/MAPK pathway. Thus, to study the mechanism of the potential agents with anti-inflammatory activities might be important in the effects of chemoprevention of chronic inflammation and carcinogenesis. The main aim of this study is to evaluate the anti-oxidant and anti-inflammatory effects about Solanum nigrum Linn water extracts (SNE). Using the xanthine oxidase activity assay, we found that the stem water extracts of SNL didn't inhibit xanthine oxidase activity, but only the leaf and green fruit water extracts of SNL demonstrate the inhibitory activity. We also used the nitrite assay to investigate the inhibition ability of SNL on the nitrite released in culture medium of LPS-stimulated RAW 264.7 cells. We found that only the leaf and green fruit water extracts of SNL effectively inhibited the nitrite excretion. Additionally, the leaf water extracts of SNL effectively inhibited the ROS production in LPS-stimulated RAW 264.7 cells, but the stem and fruit water extracts of SNL didn’t. These results suggested that the leaf water extracts of SNL might exert the anti-inflammatory actions by suppressing NO and ROS production and xanthine oxidase activity. Moreover, we used western blot to investigate the inhibition ability of the leaf water extracts on the cellular inflammatory signal transduction pathway . The data demonstrated that the leaf water extracts greatly inhibited the p38/JNK MAPK activity , and also had the ability to decrease the NF-κB (p65)/AP-1 (phospho c-jun) nuclear translocation level. Then, we used RT-PCR to assay the iNOS mRNA expression, indicating that the leaf water extracts could inhibit iNOS mRNA expression dose-dependently.

Pharmacological basis for use of Pistacia integerrima leaves in hyperuricemia and gout

Xanthine oxidase activity in human tissues and its inhibition by allopurinol (4-hydroxypyrazolo[3,4-d] pyrimidine).

Taraxacum mirabile Wagenitz, one of the endemic riches of Anatolia, is a species that has remained largely unexplored regarding its enzyme inhibition profile despite its pharmacological potential. The effects of T. mirabile aerial and root extracts, obtained at different polarities, were scrutinized in this study against two important enzymes: lactoperoxidase (LPO), which plays a vital role in the innate immune system, and xanthine oxidase (XO), which is prominently associated with hyperuricemia and oxidative stress. The aerial and root portions of the plant were extracted into fractions of varying polarities using petroleum ether, dichloromethane, ethyl acetate, and butanol. LPO was isolated from buffalo milk (881.6-fold purification, 22.5% yield, and 1249.9 EU/mg specific activity) via affinity chromatography and used in in vitro inhibition assays alongside commercial bovine XO enzyme. The results showed that the ethyl acetate fraction of the aerial part of the plant exhibited the strongest LPO inhibition (IC50: 15.60 ± 0.77 µg/mL) among the fractions. The petroleum ether fraction of both the aerial part (IC50: 11.17 ± 0.94 µg/mL) and the root part (IC50: 11.61 ± 0.59 µg/mL) had the highest inhibitory effect for the XO enzyme. These distinct inhibition profiles allow for significant insights into how plant extracts with varying polarities modulate XO and LPO enzymes. In conclusion, the significant inhibitory activity of T. mirabile extracts toward LPO and XO enzymes highlights their potential as a natural source for developing effective enzyme inhibitors, which could be useful for therapeutic applications.

Free radicals and myocardial ischemia and reperfusion injury.

The enzymes aldehyde oxidase and xanthine oxidase catalyze the oxidation of a wide range of N-heterocycles and aldehydes. These enzymes are widely known for their role in the metabolism of N-heterocyclic xenobiotics where they provide a protective barrier by aiding in the detoxification of ingested nitrogen-containing heterocycles. Isovanillin has been shown to inhibit the metabolism of aromatic aldehydes by aldehyde oxidase, but its inhibition towards the heterocyclic compounds has not been studied. The present investigation examines the oxidation of phthalazine in the absence and in the presence of the inhibitor isovanillin by partially purified aldehyde oxidase from guinea pig liver. In addition, the interaction of phthalazine with freshly prepared guinea pig liver slices, both in the absence and presence of specific inhibitors of several liver oxidizing enzymes, was investigated. ldehyde oxidase rapidly converted phthalazine into 1-phthalazinone, which was completely inhibited in the presence of isovanillin (a specific inhibitor of aldehyde oxidase). In freshly prepared liver slices, phthalazine was also rapidly converted to 1-phthalazinone. The formation of 1-phthalazinone was completely inhibited by isovanillin, whereas disulfiram (a specific inhibitor of aldehyde dehydrogenase) only inhibited 1-phthalazinone formation by 24% and allopurinol (a specific inhibitor of xanthine oxidase) had little effect. Therefore, isovanillin has been proved as an inhibitor of the metabolism of heterocyclic substrates, such as phthalazine, by guinea pig liver aldehyde oxidase, since it had not been tested before. Thus it would appear from the inhibitor results that aldehyde oxidase is the predominant enzyme in the oxidation of phthalazine to 1-phthalazinone in freshly prepared guinea pig liver slices, whereas xanthine oxidase only contributes to a small extent and aldehyde dehydrogenase does not take any part.

Uric acid cause inflammation of acute gout arthtritis and other complications. Giving chemical drugs has side effects. Flavonoids in shallots (Allium ascalonicum L.) can inhibit the xanthine oxidase enzyme as antihyperuricemia. Gingerol in red ginger (Zingiber officinale var rubrum) as antihyperuricemia with antiinflammatory effect. The research aims to determine the effectiveness of the polyherbal extract of shallots (Allium ascalonicum L.) and red ginger (Zingiber officinale var rubrum) in hyperuricemia rats. Twenty five of male rats were divided into five goups, namely negative control, positive control, (P1) 25% red ginger extract and 75% shallot, (P2) 50% red ginger extract and 50% shallot, (P3) 75% red ginger extract and 25% shallot. Potassium oxonate 250 mg / kg BW was induced intraperitoneally on the 7 th day. Uric acid has been measured on 14 th , 21 th and 28 th days. On the 15 th to 28 th days, herbs/chemical drugs were administered according to the groups. Data were analyzed using Anova Multifactorial Randomized Design, continued with Post Hoc with the Least Significant Different test. The results showed that administration of potassium oxonate, chemical drugs and combination of herbs extracts had a significant effect (p <0.05) on uric acid levels compare with negative control. The conclusion show that the administration of polyherbal shallot extract (Allium ascalonicum L.) and red ginger (Zingiber officinale var rubrum) indicates as antihyperuricemia . The most effective dose is red ginger extract 450 mg/200 g and shallot 150 mg/200 g BW/day for two weeks of administration.

To screen Vietnamese medicinal plants for xanthine oxidase (XO) inhibitory activity and to isolate XO inhibitor(s) from the most active plant. The plants materials were extracted by methanol. The active plant materials were fractionated using different organic solvents, including n-hexane, ethyl acetate, and n-butanol. Bioassay-guided fractionation and column chromatography were used to isolate compounds. The compounds structures were elucidated by analysis of spectroscopic data, including IR, MS, and NMR. Three hundreds and eleven methanol extracts (CME) belonging to 301 Vietnamese herbs were screened for XO inhibitory activity. Among these plants, 57 extracts displayed XO inhibitory activity at 100μg/mL with inhibition rates of over 50%. The extracts of Archidendron clypearia (A.clypearia), Smilax poilanei, Linociera ramiflora and Passiflora foetida exhibited the greatest potency with IC50 values below 30μg/mL. Chemical study performed on the extract of A. clypearia resulted in the isolation of six compounds, including 1-octacosanol, docosenoic acid, daucosterol, methyl gallate, quercitrin and (-)-7-O-galloyltricetiflavan. The compound (-)-7-O-galloyltricetiflavan showed the most potent XO inhibitory activity with an IC50 value of 25.5μmol/L. From this investigation, four Vietnamese medicinal plants were identified to have XO inhibitory effects with IC50 values of the methanol extracts below 30μg/mL. Compound (-)-7-O- galloyltricetiflavan was identified as an XO inhibitor from A. clypearia with IC50 value of 25.5μmol/L.

Andaliman (Zanthoxylum acanthopodium DC) is a native plant of North Sumatra province. Zanthoxylum acanthopodium is a member of Rutaceae family widely found in northern Sumatra, Indonesia. The aim of this study was to barcode Z. acanthopodium in North Sumatra province, Indonesia based on cpDNA maturase K (matK). Samples were collected in seven localities across six regions of North Sumatra province. Phylogenetic analysis was conducted using Maximum Likelihood method. The results of phylogenetic analysis indicate that Z. acanthopodium is a monophyletic group that is derived from a common ancestor. The results of the phylogenetic tree construction show that there is a grouping of accession between Z. acanthopodium species separate from other species in the Zanthoxylum genus as well as those of the Rutaceae family. The results showed that cpDNA matK marker can effectively be used as DNA barcoding to identify Z. acanthopodium.