IC50 Of Extract Research Articles

Anticancer Activity of Micromeria fruticosa and Teucrium polium Growing in Lebanon

The anticancer activities of two Lebanese plant extract Micromeria fruticose (A) and Teucrium polium (B) and their fractions were tested against MCF7 and A549 cancer cell lines using MTT assay. Separation was carried out through column chromatography and TLC analysis. Results showed that both plants possess a promising and dose dependent inhibitory activity with IC50of extract A 28.52 and 26.47 μg/ml on MCF7 and A549 cells respectively, and that of extract B was 41 and 27.9 μg/ml on MCF7 and A549 cell lines, respectively. More interestingly, combination treatment with either extracts A or B and cisplatin, significantly boosted the cytotoxic effect of cisplatin against the two cancer cell lines. Further studies are recommended to determine the active components in both plants responsible for these activities and explore their interactions at molecular level.

Antioxidant Activity of Hydro-Acetonic, Hydro-Methanolic and Aqueous Leaf and Bark Extracts of <i>Sclerocaria birrea</i> (A. Rich.) Hochst

Bioactive compounds in plants are associated with the reduction of chronic diseases. The free radical scavenging activity of different extracts of a medicinal plant, Sclerocarya birrea, has been investigated using the DPPH test, ABTS test and FRAP. Three extracts were prepared from the leaves and bark: hydro-methanolic, hydro-acetonic and aqueous. Phytochemical screening was carried out the standard methods followed by the determination of the polyphenol by Folin-Ciocalteu method. The analysis of variance (ANOVA) with the STATISTICA 7.1 and statistical significance was set at p 0.05. Evolution of percent inhibition (PI) as well as the IC50 of the extracts was obtained using the Origin Pro 8.5 software and Microsoft Excel. The results show that the bark extracts are about twice as rich in polyphenols as the leaves. With DPPH at 1.25 mg/mL, the bark has a PI of 91.04% ± 0.001% while leaves, reach 99.80% ± 0.021%. As for the ABTS test, the bark extract reached its maximum activity at 1.25 mg/mL with a PI of 99.80% ± 0.003% while leaves extract greater value of PI is 99.75 ± 0.003 at 2.5 mg/mL. With FRAP test at 1.25 mg/mL, the bark has a PI of 79.29% ± 0.005% while leaves, reach 80.33% ± 0.001%. The IC50 of the bark and leaf extracts on the smallest DPPH are 0.156 ± 0.001 mg/mL in hydro-methanol, 0.301 ± 0.00 mg/mL in hydro-acetone and 0.407 ± 0.00 mg/mL in aqueous extract. With ABTS test, the best IC50 are obtained with hydro-acetone extracts with value of 0.247 ± 0.001 mg/mL for bark and 0.248 ± 0.0005 mg/mL for leaves while in hydro-methanolic and aqueous extracts the best IC50 are respectively 0.255 ± 0.00 mg/mL and 0.463 ± 0.00 mg/mL. Using ascorbic acid as our standard, the PI was 94.86% ± 0.008% with an IC50 of 0.213 ± 0.00 mg/mL. According to these results, the reducing power of the bark is slightly higher than that of the leaves. We can say that the bark has better activity than the leaves and the alcoholic extracts have given better results than the aqueous extract.

Anticancer Activity of Ethnopharmacological Plants of Golestan Province/Iran against AGS, HT-29 and KYSE-30 Cell Lines through Promoting the Apoptosis and Immunomodulatory Effects.

The anticancer and immunomodulatory effects of medicinal plants from Golestan province, as a promising source of cancer therapy against gastrointestinal cancer cell lines, were investigated in this study. The ethanolic root/aerial part extracts of 9 medicinal plants were screened for their cytotoxicity against normal mouse fibroblast cells (L-929) and three human cancer cell lines including gastric adenocarcinoma (AGS), colorectal adenocarcinoma (HT-29), and esophagus adenocarcinoma (KYSE-30) by performing MTT assay to determine the IC50 of the extracts. The in-vitro antioxidant activity, total phenolic (TPC), and total flavonoid content (TFC) of extracts was evaluated. Flow cytometry and Real-Time PCR were used for apoptosis assay and evaluation of expression of some genes involved in cell signaling; TLR-4, AKT, ERK1/2, and NFκB. Out of the 9 plant extracts screened, Arctiumlappa root (ALR), showed the most potent cytotoxicity against AGS, KYSE-30, and HT-29 cells with IC50 values of 10, 200, and 2030 µg/mL, respectively. In addition, ALR exerts high TPC (215.8 ± 0.3 mg GAE/g), TFC (69.03 ± 0.7 mg QUE/g) and high radical scavenging activity with IC50 (1250 ‎±‎ 0.1 µg/mL) in DPPH method. Also, ALR stimulates TLR-4 signaling, increased apoptosis, and decreased cancer cell attachment to the surface compared to the untreated cells. This plant, with a strong cytotoxic effect on cancer cells as well as increased apoptosis and its effect on molecules involved in TLR4 signaling as the immunomodulatory effect can be a suitable candidate for in-vivo studies in the future for cancer therapy.

Modulation of steroid hormone synthesis by methanolic extracts of Boerhavia diffusa

Steroid hormones such as oestrogen and progesterone play multidimensional role in regulation of reproduction, metabolic and neuroendocrine functions in humans and animals. Phytoestrogens bind to oestrogen receptors and mimic the role of oestrogen, thus play a role in the management of fertility as well as therapy of hormone dependent cancers. The whole plant of Boerrhavia diffusa was collected locally from the campus of College of Veterinary and Animal sciences, Mannuthy, Thrissur. The qualitative phytochemical analysis was performed on the methanolic extracts of Boerrhavia diffusa for the presence of steroids, alkaloids, tannins, phenolic compounds, flavonoids, diterpenes, triterpenes and saponins [1]. In the present study, methanolic extract of whole plant of B. diffusa was analysed for its modulatory activity on steroidogenesis. The cytotoxicity of methanolic extract of B. diffusa was explored using MTT assay in MCF-7 cells and IC50 was calculated. The cells were also cultured in six well plates and exposed to (IC50, 2 times IC50 and half dose of IC50) for 96 hours. The culture media were collected every 48 and 96 hours, was stored at -80 °C and used for the estimation of progesterone and oestrogen by ELISA. The phytochemical analysis revealed the presence of alkaloids, glycosides, tannins and flavonoids. Through the MTT assay the IC50 of methanolic extract of B. diffusa was found to be 170 µg/mL. The methanolic extract of B. diffusa showed significant increase in the progesterone concentration with maximum increase at 340 and 170 µg/mL. There was a significant decrease in oestrogen concentration when exposed to extracts at 96 hours post treatment with maximum decrease at 340 µg/mL. Hence it could be concluded that the methanolic extract of B. diffusa down regulated oestrogen synthesis independent of the interconversion of cholesterol to progesterone. Hence the effect may be in the pathway of conversion of progesterone to oestrogen.

Total Phenol Content and Activities of Antioxidant Extracts Methanol Limes(Citrus Aurentifolia)By Uv-Vis Spectrophotometry

Total phenol levels and antioxidant activity in whole limes from Palopo, Luwu Regency, as well as the relationship between total phenol content and antioxidant activity, were determined. Maceration of dried whole lime fruit yielded the methanol extract (Citrus aurentifolia). Phytochemical screening was used to determine the presence of alkaloids, phenolic compounds, and flavonoids in the extract’s secondary metabolites. The total phenol extract was determined using the Folin-Ciocalteu method, yielding a value of 8.37 mgGAE/g. Antioxidant activity was determined using the DPPH method, and the IC50 value was 1793.06 g/mL. Total phenol content of whole fruit extract lime and IC50 show a linear relationship y = -168,11x + 3211 with a determination R2 = of 0.9780.

Skin-Related Properties and Constituents from the Aerial Parts Extract of Persicaria senticosa.

In the course of screening for cosmetic ingredients by measuring antioxidant and antiwrinkle and whitening and anti-inflammatory activities, skin-related activity was tested using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging, elastase inhibition, tyrosinase inhibition, and nitric oxide assay. Several Polygonaseae extracts were found to show potent activity. The results showed that the Persicaria senticosa methanolic extract has the 1,1­diphenyl-2­picrylhydrazyl (DPPH) and ABTS radical scavenging activities (IC50 61.0 and 17.5 μg/mL). In the elastase inhibition assay and nitric oxide assay, the IC50 of methanolic extract of Persicaria senticosa was 739.7 μg/mL and 71.8 μg/mL. The Persicaria senticosa 70% ethanolic extract partitioned with n-hexane, CH2Cl2, EtOAc, n-BuOH, and aqueous fractions. The purification of EtOAc soluble layer was by column chromatography separation and MPLC analysis of Compounds 1-7. It was identified as loliolide (1), quercetin-3-O-glucoside (2), quercetin-3-O-glucuronide (3), 4-methoxy caftraric acid (4), kaempferol-3-(6-methylglucuronide) (5), quercetin-3-(6-methylglucuronide) (6), and quercetin (7). Structure was elucidated by a combination of 1D and 2D NMR and MS spectrometry as well as comparison with reported literatures. Radical scavenging effect on DPPH, tyrosinase inhibition, and nitric oxide assay on several compounds from Persicaria senticosa was found to show potent activity. The results showed that Compound 7 has the NO assay (IC5029.7 μM). For DPPH, the IC50 of Compounds 2, 3, 5, and 7 was 39.6, 31.2, 37.0, and 22.7 μM. In tyrosinase inhibitory activity, the IC50 of Compound 7 was 14.3 μM.

Melientha suavis Pierre. Extract: Antioxidant and Sunscreen Properties for Future Cosmetic Development

The ultimate aim of this study was to evaluate antioxidant activities using DPPH and ABTS assay and antioxidant substances of Melientha suavis Pierre (Melientha) extracts with ethanol (EtOH) and subsequent partition with hexane, dichloromethane (CH2Cl2), ethyl acetate (EtOAc) and distilled water (W). The Melientha extract was assessed the sunscreen activities by sun protection factor which was compared with octyldimethyl PABA. Additionally, the stability test of Melientha extracts added in the cosmetic base-formula was investigated. The sun protection substances of Melientha extracts were determined by GC-MS analysis. Among the extract and fractions, the ethanolic extract showed the highest activities. The IC50 of Melientha extract to free radical scavenging determined by DPPH assay and ABTS assay were 53.20 ± 7.37 μg/ml and 64.17 ± 5.76 μg/ml, respectively. The total phenolic compound of ethanol extract was 149.87 ± 2.72 mg GE/g of ext and total flavonoid content was 51.60 ± 4.12 mg CE/g of ext. The sun protection factor (SPF) of ethanol extract was 26.61 ± 0.10. Cinamate and its derivative which were claim as sunscreen substance was found by GC-MS analysis of Melientha ethanol extract.

Bactris guineensis (Arecaceae) extract: Polyphenol characterization, antioxidant capacity and cytotoxicity against cancer cell lines

BACKGROUND: Lower risk of digestive tract cancer development has been associated with polyphenol intake. Bactris guineensis is an edible endemic palm that grows in Central and South America. OBJECTIVE: This study performs a phenolic characterization of Bactris guineensis and evaluates the bioactivity of this fruit. METHODS: The phenolic compounds of B. guineensis were characterized by HPLC-UV-HRMS analyses and the antioxidant activity was measured by chemical and cellular methods. Additionally, cytotoxicity of B. guineensis polyphenols was performed on 4 cancer cell lines and the pro-apoptotic effect was evaluated by flow cytometry using annexin staining. RESULTS: The major phenolic compounds of B. guineensis were proanthocyanidins. The extract IC50 for DPPH was 3.3±0.2 μg/mL and for induced intracellular ROS was 153±13 μg/mL. MTT cytotoxic assays demonstrate IC50 values between 16.6 and 24.9 μg/mL for the colon and hepatic adenocarcinomas, with high selectivity effects towards cancer cells compared to non-tumor cells. A 20 to 50% early apoptotic effect was observed in cancer cells lines by Annexin/PI staining. CONCLUSIONS: B. guineensis evidenced an important radical scavenging activity and a strong cytotoxic activity against hepatic and colorectal carcinoma cells, showing better values than procyanidin extracts from other fruits previously described.

Chemical profiling and quantification of potential active constituents responsible for the antiplasmodial activity of Cissampelos pareira

Ethnopharmacological relevanceCissampelos pareira is used traditionally in India as a remedy for the treatment of various diseases including malaria but the active ingredients responsible for antiplasmodial activity have not yet been investigated.Aim of the study: The identification and quantification of compounds responsible for antiplasmodial activity in different parts (leaf, stem and root) of C. pareira is the target of current study. Material and methodsThe hydro ethanolic parent extracts of different parts of C. pareira and fractions prepared from these extracts were evaluated against Pf3D7 (chloroquine sensitive) and PfINDO (chloroquine resistance) strains in culture to quantify the IC50 for extracts and fractions. Promising fractions of root part of plant were subjected to silica gel column chromatography to obtain pure compounds and their structures were elucidated by detailed spectroscopic analysis. Pure compounds were also tested against Pf3D7 and PfINDO strains. A rapid and simple UPLC-DAD method was developed for the identification and quantification of pharmaceutically important metabolites of C. pareira. ResultsAmong different extracts, the hydro ethanolic extract of root part of C. pareira was found most active with IC50 values (μg/ml) of 1.42 and 1.15 against Pf 3D7 and Pf INDO, respectively. Tested against Pf 3D7 the most potent fractions were root ethyl acetate fraction (IC50 4.0 μg/ml), stem water fraction (IC50 4.4 μg/ml), and root water fraction (IC50 8.5 μg/ml). Further, phytochemical investigation of active fractions of root part led to the isolation and characterization of a new isoquinoline alkaloid, namely pareirarine (8), along with five known compounds magnoflorine (5), magnocurarine (10), salutaridine (11), cissamine (13) and hayatinine (15). Hayatinine (15), a bisbenzylisoquinoline alkaloid, isolated from root ethyl acetate fraction was most promising compound with IC50 of 0.41 μM (Pf INDO) and 0.509 μM (Pf 3D7). Magnocurarine (10) and cissamine (13) were also found active with IC50 values of 12.51 and 47.34 μM against Pf INDO and 12.54 and 8.76 μM against Pf 3D7, respectively. A total of thirty compounds were detected in studied extracts and fractions, structures were assigned to 15 of these and five of these biologically important compounds were quantified. Isolation of saluteridine (11) from C. pareira and the evaluation of antiplasmodial activity of pure compound from C. pariera is disclosed for the first time. ConclusionThis study concludes that the antimalarial potential of C. pareira may be attributed to isoquinoline type alkaloids present in this plant and also provides the scientific evidence for the traditional use of this plant in treatment of malaria.

Cytotoxicity Effects of Benalu Tea Extract (Scurrula oortina) on Hella cells

Tea garden waste has been investigated in the form of parasite tea Squrula oortina Species Family: Loranthaceae from the Dempo Pagar Alam tea plantation, Lahat Regency, South Sumatra. Stem and leaf samples are then dried and powdered, then isolated with 70% water-ethanol subfraction. The extract was then dried by vacuum distillation at 70oC until a concentrated extract without ethanol was obtained. The results of the study were tea parasite extract, and the extract was tested for phytochemicals (monounsaturated fatty acids and double, theobromine derivates, quercetin derivates and Epicatekin derivates). Next, it would be taken to the Laboratory of Biological Sciences at Gajah Mada University to be tested for cytotoxic effects on Hella cells, with control positive doxorubicin. The results of the research were tea leaves extract IC50 = 2538 ug/mL and the standard of doxorubicin IC50 = 0.88 ug/mL so that it could be concluded that the parasite of the tea parasite had cytotoxic and proliferative effects.

Toxicity of Saffron Extracts on Cancer and Normal Cells: A Review Article.

Background and Aim:Medicinal plants have played an important role in human health since the Stone Age. According to WHO, 80% of Asian and African people rely on traditional medicine and medicinal plants to conserve their health. Saffron has received much attention among the herbal compounds related to cancer treatment. Methods:This review aims to provide an overview of in-vitro and in-vivo evaluation molecule mechanism for anti-tumor activity, cancer preventing and protective effects of saffron extract. The review is based on the available data accessible in PubMed, Science Direct, Google Scholar, Magiran.ir, and SID.ir databases. Results:Saffron has selective toxic and preventive effects on cancerous cells and without adverse effects on normal cells and prevents tumor formation. Saffron appears to reduce the toxic effects of anticancer drugs. Saffron has toxicity effects when used in high amounts, which are far greater than those are used in human food culture. Conclusions:Considering the observed effects of saffron on the removal of cancer cells, saffron extract can be used in the treatment and prevention of cancer after confirmation in human clinical trials. According to the high IC50 of saffron extracts in normal cells, its toxicity against non-cancerous cells is low and its use is safe. Besides, the studies suggested the cytotoxic effects of saffron on some of the more cancers, including nervous system cancer and common cancers. Further studies are required to determine the effective dose and influence of mechanism of saffron in various animal type of cancers.

Antiplasmodial activities of crude <I>Moringa oleifera</I> leaves extracts on chloroquine sensitive <I>Plasmodium falciparum</I> (3D7)

The antimalarial efficacy of crude hexane, methanol and lyophilized aqueous Moringa oleifera leaf extract was evaluated on chloroquine sensitive (CQS) strain of Plasmodium falciparum (3D7) in vitro, with a view to validate traditional use of M. oleifera as antimalarial. A dose dependent suppression of parasite growth was observed for all extracts, at microgram per mills (µg/mL) concentrations. At the lowest concentration of extract (6.25 µg/mL), the hexane, methanol and lyophilized aqueous extracts showed 63.52%, 71.42% and 60.65% suppression of plasmodium growth respectively. The extracts showed potent biological activity at the highest concentration of extract, with a strong inhibition of plasmodium growth (71.31%, 83.06% and 80.36%) observed for the hexane, methanol and lyophilized aqueous extracts. Although some extracts are observed to be more potent than others, all extracts are observed to be biologically active against the 3D7 strain of P. falciparum (Hexane extract IC50 = 3.36 µg/mL; methanol IC50 = 3.44 µg/mL and aqueous IC50 = 4.09 µg/mL respectively). The antiplasmodial activities observed may well be attributed to the presence of phenols, tannins, alkaloids and flavonoids in all solvent extracts.
 Key words: Antiplasmodial, Moringa oleifera, Phytochemical screening, Plasmodium falciparum

The investigation of cytotoxic effects of Cinnamomum zeylanicum on human breast cancer cell line (MCF-7)

Introduction: Cinnamon is one of the oldest herbal medicines that has been traditionally used as flavouring agent, insecticidal, antifungal and also as tooth powder to treat dental problems. Nowadays cinnamon is used in pharmaceutical preparations, perfume production, chemical industry and food industry as seasonings. In this study, the cytotoxicity of the cinnamon extracts towards human breast cancer cell line (MCF-7) was investigated. Methods: Cinnamomum zeylanicum was extracted using Soxhlet extraction method with methanol as the solvent, producing 7.06% of yield extract. The extract was analysed using gas chromatography/ mass spectrometry (GCMS). Results: The result showed that cinnamaldehyde was the major constituent, forming 74.14% of the total constituents. The cytotoxicity of C. zeylanicum extract against MCF-7 was determined through colorimetric MTT assay. It was demonstarted that the IC50 of extracts at 24 hours and 48 hours of incubation period was 48 µg/ml and 140 µg/ml respectively. Conclusions: The cinnamon methanolic extract possess cytotoxic effects on human breast cancer cell lines.

Xao tam phan (Paramignya trimera) methanol extract induced apoptosis in hepatocellular carcinoma HepG2 cell line in vitro

Introduction: Xao Tam Phan (Paramignya trimera) has long been used in Viet Nam as an herbal medicine for the treatment of Hepatitis, hepatocellular carcinoma, and diabetes. This study aimed to determine the anti-proliferation effect of Paramignya trimera extract (P. trimera extract) on HepG2 hepatocellular carcinoma cells.
 Methods: AlamarBlue assay was used to determine the IC50 values of P. trimera extract on HepG2 cells. Adipose-derived stem cells (ADSCs) was used as normal cell control. For apoptosis examination, P. trimera extract-treated HepG2 cells were incubated with Annexin V/Propidium iodide (PI). Then they have been analyzed their expression of Annexin-V and PI by flow cytometry. The cell nuclear degradation also was evaluated by PI/Hoechst 33342 staining assay.
 Results: Doxorubicin and P. trimera extract IC50 values on HepG2 cells were 55.13 +/- 2.028 ng/ml and 582.533 +/- 16.521 mg/ml, respectively. Those on ADSCs were 5.96 +/- 0.56 ng/ml and 268.976 +/- 19.325 mg/ml, respectively. Side effect index value (SEI) of P. trimera extract was 2.175 +/- 0.12, and the SEI of doxorubicin was 8.71 +/- 0.36. Flow cytometry analysis indicated significant apoptosis on P. trimera extract-treated HepG2 cells at a dose of 500 mg/ml (32.39 +/- 2.28% apoptotic cells, and 14.63 +/- 1.59% necrotic cells). Nuclear aggregation and degradation was seen on 500 mg/ml P. trimera treated HepG2 cells.
 Conclusion: P. trimera extract could inhibit HepG2 hepatocellular carcinoma cell proliferation by inducing apoptosis.

Ovarian cancer cells with CD133+ phenotype is more resistant against Ngai Bun Boesenbergia pandurata extract than original ovarian cancer cells

Introduction: Ovarian cancer is one of the most common cancers in women. Due to the difficulty in early detection and treatment of ovarian cancer, many research studies and clinical trials have been developed to discover more efficient therapies. Besides Western medicine, traditional medicine has gained increased interest as a research field with potential to lead to the production of marketable therapeutic products. With the diversity of tropical plants in Asia, traditional medicine has been very popular and has served as a traditional therapy for generations. The Ngai bun (Boesenbergia pandurata) root is used not only as a food spice but also in ethnomedicine. This study aimed to compare the anti-tumor activity of Boesenbergia pandurata root extract against ovarian cancer cells and CD133+ ovarian cancer cells that were enriched from the original ovarian cancer cells.
 Methods: Crude extract of Boesenbergia pandurata roots were prepared in two kinds of solvents (methanol and chloroform). The ovarian cancer cells OVP-10 were used in this study. The population of CD133+ ovarian cancer cells (CD133+OVP-10) were sorted from the OVP-10 cancer cells. Both OVP-10 cells and CD133+OVP-10 cells were treated with these crude extracts. Adiposederived stem cells (ADSCs) were used as control normal cells for all assays. The anti-tumor activity of extracts were evaluated based on the IC50 values.
 Results: Based on the IC50 index, the chloroform extract had an anti-tumor activity higher than that of methanol extract, on both OVP-10 and CD133+OPV-10 cells (IC50 of methanol and chloroform extracts were 330.1 +/- 16.9 ug/mL and 246.5 +/- 21.2 ug/mL, respectively, for OVP-10 cells; IC50 of methanol and chloroform extracts were 411.8 +/- 83.7 ug/mL and 307 +/- 9.2 ug/mL respectively, for CD133+OVP-10 cells). The results also showed that CD133+OVP-10 cells were more resistant to chloroform extract than were OVP-10 cells (307 +/- 9.2 mg/mL vs. 246.5 +/- 21.2 mg/mL, respectively, for CD133+OVP-10 vs. OVP-10 cells, p < 0.05).
 Conclusion: The chloroform extract of Boesenbergia pandurata roots displayed strong antitumor activity against ovarian cancer cells OVP-10 and CD133+OVP-10; the latter cells were found to be more resistant than the original ovarian cancer cells.

Antimicrobial activities of Allium staticiforme and Allium subhirsutum

The antibacterial and antifungal activities of the bulb and flower of Allium staticiforme and Allium subhirsutum were investigated. In addition, DPPH radical scavenging activity and total phenolic contents were determined. The results show that methanolic extracts of A. staticiforme and A. subhirsutum had antifungal activities against Candida albicans, together with a less activity level against Escherichia coli, Staphylococcus epidermidis, S. aureus, Enterecoccus faecalis, Salmonella typhimurium and Pseudomanas aeruginosa. The total phenolic contents of A. staticiforme leaf and bulb were determined as 17 and 2.4 mg of GAE/100 g, respectively. The IC50 of methanolic extracts of A. staticiforme and A. subhirsutum were also determined. In conclusion, both A. staticiforme and A. subhirsutum have antifungal activities with weak antibacterial activities. These plants have DPPH radical scavenging activities.
 Video Clip of Methodology:
 Disc Diffusion Method: 3 min 04 sec: Full Screen Alternate

Anti-platelet and anti-inflammatory properties of an ethanol-water red grape pomace extract

AbstractPrevious reports support that wine contains a mixture of micro-constituents in a proper quality and quantity that possess cardio-protective effect partly through Platelet-Activating Factor (PAF) inhibition. Grape pomace (GP) is a source of wine-like micro-constituents that may be a suitable alternative in food fortification. Limited data exist concerning their effects on thrombosis and inflammation. Therefore, the purpose of this study was to examine a grape pomace extract regarding its anti-platelet and anti-inflammatory properties. GP from four red grape varieties were extracted with 80% ethanol (GP:solvent 1:5w/v). The extract's total phenolic compounds were evaluated and the phenolic profile was performed by Ultrahigh-Performance-Liquid-Chromatography coupled to Mass-Spectrometry and the determination of fatty acids profile was performed by Gas-Chromatography. The extract's anti-platelet properties were tested in healthy volunteers’ platelet rich plasma by the light transmittance method, against three agonists: PAF, ADP and TRAP. The results expressed as IC50 values (μg of extract that cause 50% inhibition of aggregation) and EC50 values (agonist concentration that causes 50% of the maximum aggregation) in the extract's presence and absence. Concerning the extract's anti-inflammatory properties, peripheral blood mononuclear cells from healthy volunteers were pre-incubated with different extract concentrations, which were tested for their effect on cell viability, for 1 h and then stimulated with LPS for 4 h. Secretion of IL-1β and TNF-a was measured and normalized with the total cell protein. Phenolic compounds were calculated at 8.79 ± 1.17 mg gallic acid per g of GP. The most abundant ones were catechin, epicatechin and quercetin at 202.9 ± 6.9, 84.8 ± 1.5 and 83.7 ± 3.5μg per g of GP respectively. Out of the 18 fatty acids detected, the most abundant ones were palmitic, oleic, linoleic, and linolenic acid at 28.7 ± 0.1, 11.4 ± 0.01, 32.5 ± 0.07, 12.7 ± 0.005 g per 100 g of fat. The extract's IC50 was calculated at 162.1 ± 66.9, 181.2 ± 82.3 and 156.3 ± 97.5μg against PAF, ADP and TRAP, respectively. The EC50 values in the presence of 150μg extract were increased (lower platelet aggregation sensitivity) approximately at 100%, 45% and 13% against PAF, ADP and TRAP respectively, compared to EC50 values in the absence of extract. The presence of 500 and 1000μg/mL of extract reduced LPS-induced TNF-a secretion at approximately 38.2% (p = 0.04) and 6.0% (p < 0.000), respectively. Potent anti-platelet and anti-inflammatory properties are combined in a grape pomace extract. The use of its bioactive micro-constituents is likely to lead to the production of functional foods with cardioprotective properties.

Cyclooxygenase, Lipoxygenase, Nitric Oxide Synthase, Myeloperoxidase and Protease Inhibiting Activities of the Leaves and Flowers of Mikania micrantha Kunth

Aim: To study in vitro anti-inflammatory activity of the leaves and flowers of the perennial climber Mikania micrantha. Methods: Leaves and flowers of the plant was collected in February 2015. Dried and powdered plant materials was extracted with ethyl acetate at room temperature in an orbital shaker. Cyclooxygenase, 5-lipoxygenase, myeloperoxidase and nitric oxide synthase inhibiting activities of the ethyl acetate extract were tested in human monocytic cell line THP1 after activation with lipopolysaccharide. Protease inhibiting activities were tested using trypsin as the enzyme. Percentage inhibitions of enzyme activities were calculated and IC50 values were determined. Diclofenac sodium was used as the reference drug. Results: Ethyl acetate extract of the leaves as well as flowers exhibited very good myeloperoxidase, nitric oxide synthase, cyclooxygenase and lipoxygenase inhibiting activities and showed moderate effect towards protease inhibition. The IC50 of inducible nitric oxide synthase activity was 5 and 11µg respectively for leaf and flower extracts. IC50 of myeloperoxidase activity was 4µg for flower extract. The inhibition of myeloperoxidase activity by leaf extract was so high that IC50 could not be determined at the tested concentration. IC50 of cyclooxygenase activity was 25 and 89µg and lipoxygenase activity was 98 and 185µg respectively for leaf and flower extracts. The IC50 of leaf and flower extracts towards protease inhibition were 1180 and 1250 µg respectively. Conclusion: This study revealed that Mikania micrantha contains principles which can inhibit diverse enzymes of inflammatory pathway and is a potential target for the development of new anti-inflammatory drug.

PENGARUH RASIO PELARUT TERHADAP EKSTRAKSI DARI KULIT BUAH NAGA MERAH (Hylocereus polyrhizus)

This study aims to determine the effect of the ratio of solvents to extract yield, phenolics total and IC50 values ​​of red dragon fruit peel and also get the optimal solvent ratio to obtain extract yield, phenolics total and the highest or best IC50 values ​​of red dragon fruit peel extracts. The solvent ratio used there are 6 levels of the ratio: 2:1; 3:1; 4:1; 5:1; 6:1 and 7:1 (v/w). The data obtained were analyzed using a Completely Randomized Design that was applied to observations of extract yield, phenolics total and IC50 values, if the treatment had a very significant or significant effect followed by continued Tukey HSD test at 1% or 5% level. The results showed that the solvent ratio very significantly affected the extract yield, phenolics total and IC50 value of red dragon fruit peel extract. The solvent ratio of 4:1 (v/w) produced extract yield, phenolics total and IC50 values the highest or best i.e 26.22%, 71.56 ppm, and 124.62 ppm.
 Keywords : IC50, the red dragon fruit skin, solvent ratio, phenolics total

PENGARUH JENIS PELARUT TERHADAP EKSTRAKSI DARI KULIT BUAH NAGA MERAH (Hylocereus polyrhizus)

This study aims to determine the effect of the type of solvent on extract yield, total phenolics and IC50 values of the skin of red dragon fruit and also obtain the best type of solvent that provides extract rendement, total phenolics and the highest or best IC50 value for extracting red dragon fruit peels. There are 3 types of solvents used (95% ethanol, ethyl acetate and acetone: water (7: 3) with a sample-solvent ratio (5:1 v/b). The data obtained were analyzed using a Completely Randomized Design that was applied to observations of extract rendemen, phenolics total and IC50 values, if the treatment had a very significant or significant effect followed by continued BNJ test at 1% or 5% level. The results showed that the solvent ratio very significantly affected the extract rendemen, phenolics total and IC50 value of red dragon fruit peel extract. Ethanol 95% solvent is the best solvent for extracting dragon fruit peels because it has the highest yield (26.15%), total phenolic (64.75 ppm) and antioxidant activity (IC50 value) (120.53 ppm) highest or best.
 Keywords : IC50, the red dragon fruit skin, solvent ratio, extract yield, phenolics total