Search results for: kaempferol

Authors: S. B. Bhandare, K. S. Laddha

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Podphyllum hexandrum belonging to family berberidaceae has gained attention in phytochemical and pharmacological research as it shows excellent anticancer activity and has been used in treatment of skin diseases, sunburns and radioprotection. Chemically it contains lignans and flavonoids such as kaempferol, quercetin and their glycosides. Objective: To isolate and identify Kaempferol and Quercetin from Podophyllum rhizome. Method: The powdered rhizome of Podophyllum hexandrum was subjected to soxhlet extraction with methanol. This methanolic extract is used to obtain podophyllin. Podohyllin was extracted with ethyl acetate and this extract was then concentrated and subjected to column chromatography to obtain purified kaempferol and quercetin. Result: Isolated kaempferol, quercetin were light yellow and dark yellow in colour respectively. TLC of the isolated compounds was performed using chloroform: methanol (9:1) which showed single band on silica plate at Rf 0.6 and 0.4 for kaempferol and quercetin. UV spectrometric studies showed UV maxima (methanol) at 259, 360 nm and 260, 370 nm which are identical with standard kaempferol and quercetin respectively. Both IR spectra exhibited prominent absorption bands for free phenolic OH at 3277 and 3296.2 cm-1 and for conjugated C=O at 1597 and 1659.7 cm-1 respectively. The mass spectrum of kaempferol and quercetin showed (M+1) peak at m/z 287 and 303.09 respectively. 1H NMR analysis of both isolated compounds exhibited typical four-peak pattern of two doublets at δ 6.86 and δ 8.01 which was assigned to H-3’,5’ and H-2’,6’ respectively. Absence of signals less than δ 6.81 in the 1H NMR spectrum supported the aromatic nature of compound. Kaempferol and Quercetin showed 98.1% and 97% purity by HPLC at UV 370 nm. Conclusion: Easy and simple method for isolation of Kaempferol and Quercetin was developed and their structures were confirmed by UV, IR, NMR and mass studies. Method has shown good reproducibility, yield and purity.

Keywords: flavonoids, kaempferol, podophyllum rhizome, quercetin

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Authors: Ya'u Muhammad, Umar Umar A. Mallammadori, Dahiru Mansur

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Effect of kaempferol on haematological parameters in two weeks old broiler chickens with experimental Eimeria tenella infection was evaluated in this study. Sixty-day old broilers were randomly allotted into six groups (I-VI) of ten broilers each and brooded for two weeks with commercial broiler feed (vital feed®) and provided water ad libitum. At two weeks of age broilers in group 1 were neither infected nor treated. Broilers in groups II-VI were infected with Eimeria tenella sporulated oocyst (104/ml) via oral inoculation. After infection was established, broilers in groups II-IV were treated orally with 1 mg/kg, 1.5 mg/kg, and 2 mg/kg of kaempferol, respectively. Broilers in group V were treated for five days with amprolium, 1.25 g/L in drinking water. Broilers in group VI were administered normal saline, 5 ml/kg per os for five days. Five days post infection; all broilers were sacrificed by severing their jugular veins. Blood sample from each bird was collected in EDTA container for haematology. Caecal contents were harvested and used to determine the lesion score and caecal Oocyst count respectively. Data obtained was analyzed using pad prism version 5.0. Mean Packed Cell Volume (PCV), haemoglobin (Hb) concentration, and Red Blood Cell (RBC) count significantly (P < 0.05) increased in groups II, III, and IV in a dose dependent manner. Similarly, PCV, Hb concentration, and RBC count significantly (P < 0.05) increased in groups II, III, and IV when compared to VI. No significant (P > 0.05) difference in the mean values of PCV, Hb and RBC count were recorded between groups treated with kaempferol and group V. Caecal Oocyst counts and lesion scores reduced significantly (P < 0.05) in groups II, III, and IV in a dose dependent manner. It was therefore observed in this study that kaempferol improved haematological parameters and reduced Oocyst count as well as the lesion scores in broilers infected with Eimeria tenella.

Keywords: broilers, Eimeria tenella, kaempferol, lesion scores, oocyst count,

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Authors: D. Baranova, E. Neborak

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Polyphenols, particularly flavonoids like quercetin, fisetin, and kaempferol, have gained significant attention in nutrition due to their antioxidant, senolytic, and anti-inflammatory properties. These compounds are commonly found in various plant-based foods and are represented by diverse subclasses, each with unique health benefits. Understanding their absorption, metabolism, and bioactivity within the human body is crucial for unlocking their full potential. Quercetin, for instance, exists in multiple forms, impacting its solubility and absorption in the intestine. Its intake, often derived from sources like apples, is affected by cooking methods, with medium heat retaining its potency. Fisetin, also present in fruits and vegetables, demonstrates neuroprotective qualities and stability under varied conditions compared to quercetin. Similarly, kaempferol, found in fruits and vegetables, displays antioxidative effects but is influenced by cooking techniques, with specific methods preserving its polyphenolic content better. Overall, these polyphenols offer promising health benefits, yet their optimal dosage and specific dietary recommendations warrant further research to harness their full nutritional potential.

Keywords: polyphenols, flavonoids, absorption, quercetin, kaempferol, fisetin, senolytics, absorption, cooking method

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Authors: Palaniyandi Karuppaiya, Hsin Sheng Tsay, Fang Chen

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Medicinal herbs do represent a huge and noteworthy reservoir for novel anticancer drugs discovery. Dysosma pleiantha (Hance) Woodson (Berberidaceae), one of the oldest traditional Chinese medicinal herb, highly prized by the mountain tribes of Taiwan and China for its medicinal properties contained pharmaceutically important antitumor compounds podophyllotoxin, quercetin and kaempferol. Among lignans, podophyllotoxin is an active antitumor compound and has now been modified to produce clinically useful drugs etoposide and teniposide. In recent years, natural populations of D. peliantha have declined considerably due to anthropogenic activities such as habitat destruction and commercial exploitation for medicinal applications. As to its overall conservation status, D. pleiantha has been ranked as threatened on the China Species Red List. In the present study, an efficient in vitro callus culture system of D. pleiantha was established on Gamborg’s medium with various combinations and concentrations of different auxins and cytokinins under dark condition. Best callus induction was recorded in 2 mg/L 2, 4 - Dichlorophenoxyacetic acid (2,4-D) along with 0.2 mg/L kinetin and the maximum callus proliferation was achieved at 1 mg/L 2,4-D. Among the explants tested, maximum callus induction (86 %) was achieved from tender leaves. Hence, in subsequent experiments, leaf callus was further investigated for suitable callus biomass and production level of anticancer compounds under the influence of different additives. A maximum fresh callus biomass (8.765 g) was recorded in callus proliferation medium contained 500 mg/L casein hydrolysate. High performance liquid chromatography results revealed that the addition of different concentrations of peptone (1, 2 and 4 g/L) in callus proliferation medium enhanced podophyllotoxin (16 fold), quercetin (12 fold) and kaempferol (5 fold) accumulation than control. Thus, the established in vitro callus culture under the influence of different additives may offer an alternative source of enhanced production of podophyllotoxin, kaempferol and quecertin without harming natural plant population.

Keywords: dysosma pleiantha, kaempferol, podophyllotoxin, quercetin

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Authors: Iman A. A. Kassem, Ayman A. Farghaly, Zeinab M. Hassan, Farouk R. Melek, Neveen S. Ghaly

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Lipidium sativum L, an annual herb that grows to 50 cm, is known as an important member of family Brassicaceae. Besides its nutritional value, the seeds were widely used in folk medicine for treatment of cough, asthma, and headache. It was also reported to possess hypocholesterolemic, anti-inflammatory, antidiarrheal, antimicrobial and anticancer activities. In this study, the genoprotective properties of L. sativum seed methanolic extract (LSME) were evaluated in vivo. Three groups of mice were given LSME for five consecutive days at the three dose levels 25, 50 and 100 mg/kg b.wt. The three groups were then injected intraperitoneally with cyclophosphamide at a dose of 20 mg/kg b.wt. to induce DNA damage. A group received only cyclophosphamide (20 mg/kg b.wt.) served as control. LSME significantly inhibited the DNA aberrations in mice caused by cyclophosphamide in a dose-dependent manner in the two groups that received LSME at 50 and 100 mg/kg b.wt. dose levels. The chromosomal aberrations' inhibitory indices were calculated as 18 and 31 in mice bone marrow cells and 27 and 48 in mice spermatocytes, respectively. Phytochemical examination carried out by us revealed that flavonoids were the main chemical constituents of LSME. The major flavonoids kaempferol, kaempferol-3-O-rhamnoside, kaempferol-3-O-glucoside, quercetin, and quercetin-3-O-glucoside were isolated and characterized. It was concluded that the genoprotective effect of LSME might be attributed to the presence of flavonoids which are well-known for their antioxidant properties.

Keywords: cyclophosphamide, flavonoids, genoprotective effect, Lepidium sativum

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Authors: Olga St. Tsiftsoglou, Diamanto M. Lazari, Eugene L. Kokkalou

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Most of Alyssum species of Brassicaceae family have been mainly studied for their contribution in ecology. In this study, A. alyssoides was examined for its chemical substitutes. The methanol extract of its aerial parts was fractionated with liquid-liquid extraction (distribution) with four different solvents of increasing polarity: diethyl ether, ethyl acetate, 1-butanol and water. The diethyl ether and ethyl acetate extracts were further studied for their chemical composition. So far, secondary metabolites which belong to phenolics were isolated by using several chromatographic methods (C.C. and HPLC) and were identified by using spectroscopic methods (UV/Vis, NMR and MS): two phenolic acids (p-hydroxy-benzoic acid and 3-methoxy-4-hydroxy-benzoic acid (vanillic acid)), and five flavonoids, which are derivatives of flavonol: kaempferol 3-O-β-D-glucopyranoside (astragalin), kaempferol 3-O-(6′′-α-L-rhamnopyranosyl)-β-D-glucopyranoside (nicotiflorin), quercetin 3-O-β-D-glucopyranoside (isoquercetin), isorhamnetin-3-O-β-D-glucopyranoside, and isoramnetin 3-O-(6′′-α-L-rhamnopyranosyl)-β-D-glucopyranoside (narcissin).

Keywords: Alyssum, chemical substitutes, flavonoids, phenolic acids

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Authors: Silvia Ximena Barrios, Diego Armando Villamizar Mantilla, Raquel Elvira Ocazionez, , Elena E. Stashenko, María Pilar Vinardell, Jorge Luis Fuentes

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Introduction: Skin overexposure to solar radiation has been a serious public health concern, because of its potential carcinogenicity. Therefore, preventive protection strategies using photoprotective agents are critical to counteract the harmful effect of solar radiation. Plants may be a source of photoprotective compounds that inhibit cellular mutations involved in skin cancer initiation. This work evaluated the photoprotective and antigenotoxic effects against ultraviolet B (UVB) radiation of a mixture of Posoqueria latifolia flower extract and Kaempferol (MixPoKa). Methods: The photoprotective efficacy of MixPoka (Posoqueria latifolia flower extract 250 μg/ml and Kaempferol 349.5 μM) was evaluated using in vitro indices such as sun protection factor SPFᵢₙ ᵥᵢₜᵣₒ and critical wavelength (λc). The MixPoKa photostability (Eff) at human minimal erythema doses (MED), according to the Fitzpatrick skin scale, was also estimated. Cytotoxicity and genotoxicity/antigenotoxicity were studied in MRC5 human fibroblasts using the trypan blue exclusion and Comet assays, respectively. Kinetics of the genetic damage repair post irradiation in the presence and absence of the MixPoka, was also evaluated. Results: The MixPoka -UV absorbance spectrum was high across the spectral bands between 200 and 400 nm. The UVB photoprotection efficacy of MixPoka was high (SPFᵢₙ ᵥᵢₜᵣₒ = 25.70 ± 0.06), showed wide photoprotection spectrum (λc = 380 ± 0), and resulted photostable (Eff = 92.3–100.0%). The MixPoka was neither cytotoxic nor genotoxic in MRC5 human fibroblasts; but presented significant antigenotoxic effect against UVB radiation. Additionally, MixPoka stimulate DNA repair post-irradiation. The potential of this phytochemical mixture as sunscreen ingredients was discussed. Conclusion: MixPoka showed a significant antigenotoxic effect against UVB radiation and stimulated DNA repair after irradiation. MixPoka could be used as an ingredient in a sunscreen cream.

Keywords: flower extract, photoprotection, antigenotoxicity, cytotoxicity, genotoxicit

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Authors: Mai M. Farid, Mona El-Shabrawy, Sameh R. Hussein, Ahmed Elkhateeb, El-Said S. Abdel-Hameed, Mona M. Marzouk

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Asphodelus aestivus Brot. Is a wild plant distributed in Egypt and is considered one of the five Asphodelus spp. from the family Asphodelaceae; it grows in dry grasslands and on rocky or sandy soil. The chemical components of A. aestivus flowers extract were analyzed using different chromatographic and spectral techniques and led to the isolation of two anthraquinones identified as emodin and emodin-O-glucoside. In addition to, five flavonoid compounds;kaempferol,Kaempferol-3-O-glucoside,Apigenin-6-C-glucoside-7-O-glucoside (Saponarine), luteolin 7-O-β-glucopyranoside, Isoorientin-O-malic acid which is a new compound in nature. The LC-ESI-MS/MS analysis of the flower extract of A. aestivus led to the identification of twenty- two compounds characterized by the presence of flavones, flavonols, and flavone C-glycosides. While GC/MS analysis led to the identification of 24 compounds comprising 98.32% of the oil, the major components of the oil were 9, 12, 15-Octadecatrieoic acid methyl ester 28.72%, and 9, 12-Octadecadieroic acid (Z, Z)-methyl ester 19.96%. In vitro cytotoxic activity of the aqueous methanol extract of A. aestivus flowers against HEPG2, HCT-116, MCF-7, and A549 culture was examined and showed moderate inhibition (62.3±1.1)% on HEPG2 cell line followed by (36.8±0.2)% inhibition on HCT-116 and a weak inhibition (5.7± 0.0.2) on MCF-7 cell line followed by (4.5± 0.4) % inhibition on A549 cell line and this is considered the first cytotoxic report of A. aestivus flowers.

Keywords: Anthraquinones, Asphodelus aestivus, Cytotoxic activity, Flavonoids, LC-ESI-MS/MS

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Authors: V. Yurina, H. Sujuti, E. Rahmani, A. R. Nopitasari

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Breast cancer has the highest prevalence cancer in women. Moringa leaves (M. oleifera) contain quercetin, kaempferol, and benzyl isothiocyanate which can enhance induction of apoptosis. This research aimed to study the role of the leaf extract of Moringa to increase apoptosis in breast cancer cell line, MCF-7 cells. This research used in vitro experimental, post-test only, control group design on breast cancer cells MCF-7 in vitro. Moringa leaves were extracted by maceration method with ethanol 70%. Cells were treated with drumstick leaves extract on 1100, 2200, and 4400 μg/ml for Hsp27 and caspase-9 expression (immunocytochemistry) and apoptosis (TUNEL assay) test. The results of this study found that the IC50 2200 µg/ml. Moringa leaves extract can significantly increase the expression of caspase-9 (p<0.05) and decreased Hsp 27 expression (p<0.05). Moreover it can increase apoptosis (p<0.05) significantly in MCF-7 cells. The conclusion of this study is Moringa leaves extract is able to increase the expression of caspase-9, decrease Hsp27 expression and increase apoptosis in breast cancer cell-line MCF-7.

Keywords: apoptosis, breast cancer, caspase-9, Hsp27, Moringa oleifera

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Authors: Xiran Wang, Johanna Trinkl, Thomas Hoffmann, Wilfried Schwab

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Malonyltransferases (MATs) are enzymes that play a key role in the biosynthesis of secondary metabolites in plants, such as flavonoids and anthocyanins. As a kind of flavonoid-rich fruit, strawberries are an ideal model to study MATs. From Goodberry metabolome data, in the hybrid generation of 2 strawberries various, Fragaria × ananassa cv. 'Senga Sengana' and 'Candonga', we found the malonylated flavonoid concentration is significantly higher in 'Senga Sengana' compared with 'Candonga'. Therefore, we aimed to identify and characterize the malonyltransferases responsible for the different malonylated flavonoid concentrations in two different strawberry cultivars. In this study, we have found 6 MATs via genome mapping, metabolome analysis, gene cloning, and enzyme assay from strawberries, which catalyzed the malonylation of flavonoid substrates: quercetin-3-glucoside, kaempferol-3-glucoside, pelargonidin-3-glucoside, and cyanidin-3-glucoside. All four compounds reacted with FaMATs to varying degrees. These MATs have important implication into strawberries’ flavonoid biosynthesis, and also provide insights into insights into flavonoid biosynthesis, potential applications in agriculture, plant science, and pharmacy, and information on the regulation of secondary metabolism in plants.

Keywords: malonyltransferase, strawberry, flavonoid biosynthesis, enzyme assay

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Authors: Stephen Adeniyi Adefegha, Vitor Mostardeiro, Vera Maria Morsch, Ademir F. Morel, Ivana Beatrice Manica Da Cruz, Sabrina Somacal Maria Rosa Chitolina Schetinger

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Moringa stenopetala is often consumed as food and used in folkloric medicine for the management of several diseases. Purpose: This study was set up in order to assess the effect of aqueous extract of Moringa stenopetala on cell viability and oxidative stress biomarkers in BV-2 microglial cells. Aqueous extracts of M. stenopetala were prepared, lyophilized and reconstituted in 0.5% dimethylsulphoxide (DMSO). Cells were treated with M. stenopetala extracts (0.1 - 100 µg/ml) for cell viability and nitric oxide (NO) production tests. However, M. stenopetala extract (50 µg/ml) was used in the treatment of cells for the determination of protein carbonyl content and reactive oxygen species (ROS) level. Incubation of BV-2 microglia cell with M. stenopetala extract maintained cell viability, diminished NO and ROS levels, and reduced protein carbonyl contents Chlorogenic acid, rutin, kaempferol and quercetin derivatives were the main phenolic compounds identified in M. stenopetala leaf extract. These phenolic compounds present in M. stenopetala may be responsible for the mitigation of oxidative stress in BV-2 microglial cells.

Keywords: oxidative stress, BV-2 microglial cell, Moringa stenopetala, cell viability, antioxidant

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Authors: Divya Choudhary, Hariprasad P., S. N. Naik

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This study examines a low-alcohol nutra-beverage made with shatavari, a plant commonly used in traditional medicine. During fermentation, the addition of a specific strain of yeast affected the beverage's properties, including its pH level, yeast count, ethanol content, and antioxidant, phenolic, and flavonoid levels. We also analyzed the beverage's storage and shelf life. Despite its bitter taste, the low alcohol content of the beverage made it enjoyable to drink and visually appealing. Our analysis showed that the optimal time for fermentation was between the 14th and 21st day when the beverage had ideal levels of sugar, organic acids, and vitamins. The final product contained fructose and citric acid but not succinic, pyruvic, lactic, or acetic acids. It also contained vitamins B2, B1, B12, and B9. During the shelf life analysis, we observed changes in the beverage's pH, TSS, and cfu levels, as well as its antioxidant activity. We also identified volatile (GC-MS) and non-volatile compounds (LC-MS/MS) in the fermented product, some of which were already present in the Shatavari root. The highest yield of product contained the maximum concentration of antioxidant compounds, which depended on both the pH and the microorganisms' physiological status. Overall, our study provides insight into the properties and potential health benefits of this Nutra-beverage.

Keywords: antioxidants, fermentation, volatile compounds, acetonin, 1-butanol, non-volatile compounds, Shatavarin V, IX, kaempferol

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Authors: Nabil Ghedadba, Abdessemed Samira, Leila Hambaba, Sidi Mohamed Ould Mokhtar, Nassima Fercha, Houas Bousselsela

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The overall objective of this study was to evaluate in vitro the hemostatic activity of secondary metabolites (polyphenols, flavonoids, and tannins) of Marrubium vulgare leaves, aromatic plant widely used in traditional medicine for the treatment of asthma, cough, diabetes (by its effect on the pancreas to secrete insulin), heart disease, fever has a high efficiency as against inflammation. Qualitative analysis of the aqueous extract (AQE) by thin layer chromatography revealed the presence of quercetin, kaempferol and rutin. Quantification of total phenols by Folin Ciocalteu method and flavonoids by AlCl3 method gave high values with AQE: 175±0.80 mg GAE per 100g of the dry matter, 23.86±0.36 mg QE per 100g of dry matter. Moreover, the assay of condensed tannins by the vanillin method showed that AQE contains the highest value: 16.55±0.03 mg e-catechin per 100 g of dry matter. Assessment of hemostatic activity by the plasma recalcification method (time of Howell) has allowed us to discover the surprising dose dependent anticoagulant effect of AQE lyophilized from leaves of M. vulgare. A positive linear correlation between the two parameters studied: the content of condensed tannins and hemostatic activity (r=0.96) were used to highlight a possible role of these compounds that are potent vasoconstrictor activity in hemostatic. From these results we can see that Marrubium vulgre could be used for the treatment of health.

Keywords: Marrubium vulgare L., aqueous extract, phenolic compounds dosing, hemostatic activity, condensed tannins

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Authors: E. Kayitesi, S. Moyo, V. Mavumengwana

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Cleome gynandra (spider plant) is an African green leafy vegetable categorized as an indigenous, underutilized and has been reported to contain essential phenolic compounds. Phenolic compounds play a significant role in human diets due to their proposed health benefits. These compounds however may be affected by different processing methods such as cooking and drying. Cleome gynandra was subjected to boiling, steam blanching, and drying processes and analysed for Total Phenolic Content (TPC), Total Flavonoid Content (TFC), antioxidant activity and flavonoid composition. Cooking and drying significantly (p < 0.05) increased the levels of phenolic compounds and antioxidant activity of the vegetable. The boiled sample filtrate exhibited the lowest TPC followed by the raw sample while the steamed sample depicted the highest TPC levels. Antioxidant activity results showed that steamed sample showed the highest DPPH, FRAP and ABTS with mean values of 499.38 ± 2.44, 578.68 ± 5.19, and 214.39 ± 12.33 μM Trolox Equivalent/g respectively. An increase in quercetin-3-rutinoside, quercetin-rhamnoside and kaempferol-3-rutinoside occurred after all the cooking and drying methods employed. Cooking and drying exerted positive effects on the vegetable’s phenolic content, antioxidant activity as a whole, but with varied effects on the individual flavonoid molecules. The results obtained help in defining the importance of African green leafy vegetable and resultant processed products as functional foods and their potential to exert health promoting properties.

Keywords: Cleome gynandra, phenolic compounds, cooking, drying, health promoting properties

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Authors: Gonzalo Ortiz de Elguea-Culebras, Raúl Sánchez-Vioquea, Adela Mena-Morales, Manuel Alaiz, Enrique Melero-Bravo, Esteban García-Romero, Javier Vioque, Lourdes Marchante-Cuevas, Julio Girón-Calle

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Saffron (Crocus sativus L.) is a highly valued crop for the manufacture of spice that consists of the dried stigma of the flowers. This is in contrast to other underutilized parts of the saffron plant as leaves, which represent abundant biomass whose use might help to enhance the sustainability of the saffron crop. Saffron leaves contain significant amounts of phenolic compounds, 7.8 equivalent grams of gallic acid per 100g of extract, and are very promising compounds in terms of exploring novel uses of saffron leaves. Given that phenolic compounds have numerous effects on cancer-related biological pathways, we have investigated the in vitro antiproliferative effect of saffron leaf polyphenols against human colon adenocarcinoma cells (Caco-2). Polyphenols were extracted from leaves with 70% ethanol, defatted with hexane, and purified by solid phase extraction using C18 silica gel and then silica gel 60. Analysis of polyphenols was performed by HPLC-ESI-MS. Di-, tri-, and tetrahexosides of quercetin, kaempferol, and isorhamnetin, as well as C-hexosides like isoorientin and vitexin, were tentatively identified. Polyphenols strongly inhibited the proliferation of Caco-2 cells, which is consistent with model studies in which several of the polyphenols identified in saffron leaves have demonstrated their potential as chemopreventive agents in cancer. Due to the low profitability that saffron leaf currently represents, we consider these results very encouraging and that this by-product deserves further investigation as a potential source of active molecules against colorectal cancer.

Keywords: saffron leaves, agricultural by-products, polyphenols, antiproliferative effect, human colon adenocarcinoma cells

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Authors: Ajay Kumar, Satwinderjeet Kaur

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Onosma bracteata Wall. (Boraginaceae), is known to be a medicinal plant, useful in the treatment of body swellings, abdominal pain and urinary calculi, etc. The present study focused on the radical scavenging and cancer growth inhibitory properties of isolates from O. bracteata. Obea fraction demonstrated noticeable free radical scavenging ability along with antiproliferative activity in human osteosarcoma MG-63, human neuroblastoma IMR-32, and human lung cancer A549 cell lines using MTT assay with GI50 values of 88.56, 101.61 and 112.7 μg/ml, respectively. The scanning electron and confocal microscopy studies showed morphological alterations including nuclear condensation and formation of apoptotic bodies in osteosarcoma MG-63 cells. Obea fraction in osteosarcoma MG-63 cells augmented the reactive oxygen species (ROS) level and decreased the mitochondrial membrane potential. Flow cytometry analysis revealed the Obea treated cells to be arrested in the G0/G1 phase in a dose dependent manner supported by the observed increase in the early apoptotic cell population. Western blotting analysis showed that the expression of p-NF-kB, COX-2, p-Akt, and Bcl-xL decreased whereas, the expression of GSK-3β, p53, caspase-3 and caspase-9 proteins increased. The downregulation of Bcl-2, Cyclin E, CDK2 and mortalin gene expression and upregulation of p53 genes was unfolded in RT-qPCR studies. The presence of catechin, kaempferol, Onosmin A and epicatechin, as revealed in high-performance liquid chromatography (HPLC) studies, contributes towards the chemopreventive potential of O. bracteata which can be tapped for chemotherapeutic use.

Keywords: apoptosis, confocal microscopy, HPLC, mitochondria membrane potential, reactive oxygen species

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Authors: R. Raudonis, L. Raudonė, V. Janulis, P. Viškelis

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Pharmacological and clinical studies demonstrated that phenolic compounds particularly flavonoids and phenolic acids are responsible for a wide spectrum of therapeutic activities. Flavonoids and phenolic acids are regarded as natural antioxidants that play an important role in protecting cells from oxidative stress. Qualitatively prepared dry extracts possess high stability and concentration of bio active compounds, facility of standardization and quality control. The aim of this work was to determine the phenolic and antioxidant profiles of Hippophaë rhamnoides L., Betula pendula Roth., Tilia cordata Mill., Sorbus aucuparia L. leaves dry extracts and to identify markers of antioxidant activity. Extracts were analyzed using high-performance liquid chromatography (HPLC) with FRAP post-column assay. Dry extracts are versatile forms possessing wide area of applications, final product ensure consistent phytochemical and functional properties. Seven flavonoids: rutin, hyperoside, isorhamnetin 3-O-rutinoside, isorhamnetin 3-O-glucoside, quercetin, kaempferol, isorhamnetin were identified in dry extract of Hippophaë rhamnoides L. leaves. Predominant compounds were flavonol glycosides which were chosen as markers for quantitative control of dry extracts. Chlorogenic acid, hyperoside, rutin, quercetin, isorhamnetin were prevailing compounds in Betula pendula Roth. leaves extract, whereas strongest ferric reducing activity was determined for chlorogenic acid and hyperoside. Notable amounts of protocatechuic acid and flavonol glycosides, rutin, hyperoside, quercitrin, isoquercitrin were identified in the chromatographic profile of Tilia cordata Mill. Neochlorogenic and chlorogenic acids were significantly dominant compounds in antioxidant profile in dry extract of Sorbus aucuparia L. leaves. Predominant compounds of antioxidant profiles could be proposed as functional markers of quality of phenolic rich raw materials. Dry extracts could be further used for manufacturing of pharmaceutical and nutraceuticals.

Keywords: dry extract, FRAP, antioxidant activity, phenolic

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Authors: Amos Sunday Onikanni, Bashir Lawal, Babatunji Emmanuel Oyinloye, Gomaa Mostafa-Hedeab, Mohammed Alorabi, Simona Cavalu, Augustine O. Olusola, Chih-Hao Wang, Gaber El-Saber Batiha

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The increasing global burden of diabetes mellitus has called for the search for a therapeutic alternative that offers better activities and safety than conventional chemotherapy. Herein, we evaluated the neuroprotective and antioxidant properties of different fractions (ethyl acetate, N-butanol and residual aqueous) of Clompanus pubescens leaves in streptozotocin (STZ)-induced diabetic rats. Our results revealed a significant elevation in the levels of blood glucose, pro-inflammatory cytokines, lipid peroxidation, neuronal activities of acetylcholinesterase, butyrylcholinesterase, nitric oxide, epinephrine, norepinephrine, and Na+/K+-ATPase in diabetic non treated rats. In addition, decreased levels of enzymatic and non-enzymatic antioxidants were observed. Treatment with different fractions of C. pubescens leaves resulted in a significant reversal of the biochemical alteration and improved the neurocognitive deficit in STZ-induced diabetic rats. However, the ethyl-acetate fraction demonstrated higher activities than the other fractions and was characterized for its phytoconstituents, revealing the presence of Gallic acid (713.00 ppm), catechin (0.91 ppm), ferulic acid (0.98 ppm), rutin (59.82 ppm), quercetin (3.22 ppm) and kaempferol (4.07 ppm). Our molecular docking analysis revealed that these compounds exhibited different binding affinities and potentials for targeting BChE/AChE/ IL-1 β/Na+-K+-ATPase. However, only Kampferol and ferulic exhibited good drug-like, ADMET, and permeability properties suitable for use as a neuronal drug target agent. Hence, the ethyl-acetate fraction of C. pubescent leaves could be considered a source of promising bioactive metabolite for the treatment and management of cognitive impairments related to type II diabetes mellitus.

Keywords: diabetes mellitus, neuroprotective, antioxidant, pro-inflammatory cytokines

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Authors: Daniela Nedeltcheva-Antonova, Kamelia Getchovska, Vera Deneva, Stanislav Bozhanov, Liudmil Antonov

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Rosa damascena Mill. (Damask rose) is one of the most important plants belonging to the Rosaceae family, with a long historical use in traditional medicine and as a valuable oil-bearing plant. Many pharmacological effects have been reported from this plant, including anti-inflammatory, hypnotic, analgesic, anticonvulsant, anti-depressant, antianxiety, antitussive, antidiabetic, relaxant effects on tracheal chains, laxative, prokinetic and hepatoprotective activities. Pharmacological studies have shown that the various health effects of R. damascena flowers can mainly be attributed to its large amount of polyphenolic components. Phenolics possess a wide range of pharmacological activities, such as antioxidants, free-radical scavengers, anticancer, anti-inflammatory, antimutagenic, and antidepressant, with flavonoids being the most numerous group of natural polyphenolic compounds. According to the technological process in the production of rose concrete (solvent extraction with non-polar solvents of fresh rose flowers), it can be assumed that the resulting plant residue would be as rich of polyphenolics, as the plant itself, and could be used for the development of novel products with promising health-promoting effect. Therefore, an optimisation of the extraction procedure of the by-product from the rose concrete production was carried out. An assay of the extracts in respect of their total polyphenols and total flavonoids content was performed. HPLC analysis of quercetin and kaempferol, the two main flavonoids found in R. damascena, was also carried out. The preliminary results have shown that the flavonoid content in the rose extracts is comparable to that of the green tea or Gingko biloba, and they could be used for the development of various products (food supplements, natural cosmetics and phyto-pharmaceutical formulation, etc.). The fact that they are derived from the by-product of industrial plant processing could add the marketing value of the final products in addition to the well-known reputation of the products obtained from Bulgarian roses (R. damascena Mill.).

Keywords: gas chromatography-mass-spectromrtry, dry extract, flavonoids, Rosa damascena Mill

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Authors: A. H. Taghdisi, M. Mirmohammadi, S. Kamali

Abstract:

Evening primrose (Oenothera biennis L.) is a biennial and herbaceous and one of the most important species of medicinal plants in the world. due to the production of unsaturated fatty acids such as linoleic acid, alpha-linolenic acid, etc. in its seeds and roots, and compounds such as kaempferol in its leaves, Evening primrose has important medicinal efficiency such as reducing premenstrual problems, acceleration of wound healing, inhibiting platelet aggregation, sedation of cardiovascular diseases, and treatment of viral infections. The sap of the plant is used to treat warts, and the plant itself is used as a charm against mental and spiritual diseases and poisonous animals. Its leaves have significant antibacterial activity against yellow staphylococci. It is also used in the treatment of poisoning, especially the toxication caused by the consumption of alcoholic beverages, in the treatment of arteriosclerosis and diseases caused by liver cell insufficiency. Low germination and production speed are the problems of evening primrose growth and propagation. In the present study, extracts were obtained from four components (flowers, stems, seeds, leaves) of the evening primrose plant using the Soxhlet apparatus. To measure the antibacterial properties against MDR bacteria, microbial methods, including dilution, cultivation on a plate containing nutrient agar culture medium, and disc diffusion in agar, were performed using Staphylococcus aureus and Escherichia coli bacteria on all four extracts. The maximum antibacterial activity related to the dilution method was obtained in all extracts. In the plate culture method, antibacterial activity was obtained for all extracts in the nutrient agar medium. The maximum diameter of the non-growth halo was obtained in the disc diffusion method in agar in the leaf extract. The statistical analysis of the microbial part was done by one-way ANOVA test (SPSS). By comparing the amount of omega-3 in extracts of Iranian and foreign oils available in the market and the extracts extracted from evening primrose plant samples with gas chromatography, it is shown that the stem extract had the most omega-3 (oleic acid) and compared to the extract of the mentioned oils, it had the highest amount of omega-3 overall. Also, the amount of omega-3 in the extract of Iranian oils was much higher than in the extract of foreign oils. It should be noted that the extract of foreign oils had a more complete composition of omega-3 than the extract of Iranian oils.

Keywords: antibacterial activity, MDR bacteria, evening primrose, omega-3

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Authors: Satria B. Mahathma, Asri Hendrawati

Abstract:

Background: Diabetes mellitus (DM) is a metabolic disorder syndrome characterized by chronic hyperglycemia. The number of people with diabetes rose from 108 million in 1980 to 422 million in 2014. In general, almost 90% of the prevalence of DM is type 2 DM which marked by insulin resistance and decreased receptor sensitivity. Aside from conventional antidiabetic therapy, the utilization of medicinal plants as alternative medicine has beneficial effects in diabetic patients. Flavonoid contents in radish leaves such as quercetin, pelargonidin, and kaempferol are thought to have antidiabetic activity on decreasing blood glucose levels by tricyclic nucleotide modulation of pancreatic beta cells and ameliorating insulin resistance. This study aimed to determine the effect of variant concentration of radish leaves ethanol extract on blood glucose levels in diabetic rats. Method: This study used pretest-posttest control group design by using 16 male Wistar rats which were induced type-2 diabetic by streptozotocin 60 mg/kg BW-nicotinamide 120 mg/kg BW intraperitoneally. Rats who had developed type-2 DM later divided randomly into 4 groups; negative control received placebo, positive control received glibenclamide 5 mg/kg BW/day, rats intervention I and intervention II received 100% and 50% of radish leaves ethanol extract, respectively. Treatments were administered orally for four weeks. The blood glucose levels were measured using the Enzymatic Colorimetric Test “GOD-PAP”. Data were analyzed by the dependent t-test for pretest-posttest intervention difference and one-way ANOVA followed by post hoc test to determine the significant difference of each treatment to obtain the significant data. Result: The result revealed that intervention group had lower blood glucose levels mean than control group which the lowest was intervention II group (negative control: 540,9 ± 191,7 mg/dl, positive control: 494, 97 ± 64,91 mg/dl, intervention I: 301,92 ± 165,70 mg/dl, and intervention II group: 276,1 ± 139,02 mg/dl. Intervention II group had the highest antidiabetic activity, followed by the intervention I group with the amount of decrease in blood glucose levels were -151,85 ± 77,43 mg/dl and -11,08 ± 186,62 mg/dl, however negative and positive control group didn’t have antidiabetic activity. The dependent t-test result showed there is a significant difference in decreasing blood glucose levels in the intervention II pretest-posttest intervention (p=0,03) while the other group didn’t. Data analyzed by one-way ANOVA also revealed the intervention II group significantly declined blood glucose levels compared to the negative and positive control group (p = 0,033 and p=0,032, respectively). Conclusion: There is a significant effect of radish leaves ethanol extract on blood glucose levels in streptozotocin-nicotinamide-induced diabetic rats with the optimal therapeutic effect at a concentration of 50%.

Keywords: blood glucose levels, medicinal plant, radish leaves, type-2 diabetes mellitus

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Authors: Elena Ionescu, Elena Iacob, Marie-Louise Ionescu, Carmen Elena Tebrencu, Oana Teodora Ciuperca

Abstract:

Echinacea species (Asteraceae) has received a global attention because it is widely used for treatment of cold, flu and upper respiratory tract infections. Echinacea species contain a great variety of chemical components that contribute to their activity. The most important components responsible for the biological activity are those with high molecular-weight such as polysaccharides, polyacetylenes, highly unsaturated alkamides and caffeic acid derivatives. The principal factors that may influence the chemical composition of Echinacea include the species and the part of plant used (aerial parts or roots ). In recent years the market for Echinacea has grown rapidly and also the cases of adultery/replacement especially for Echinacea root. The identification of presence or absence of same biomarkers provide information for safe use of Echinacea species in food supplements industry. The aim of the study was the preliminary evaluation and fingerprinting by UV-VISIBLE spectroscopy of biomarkers in terms of content in phenolic derivatives of some Echinacea species (E. purpurea, E. angustifolia and E. pallida) for identification and authentication of the species. The steps of the study were: (1) samples (extracts) preparation from Echinacea species (non-hydrolyzed and hydrolyzed ethanol extracts); (2) samples preparation of reference substances (polyphenol acids: caftaric acid, caffeic acid, chlorogenic acid, ferulic acid; flavonoids: rutoside, hyperoside, isoquercitrin and their aglycones: quercitri, quercetol, luteolin, kaempferol and apigenin); (3) identification of specific absorption at wavelengths between 700-200 nm; (4) identify the phenolic compounds from Echinacea species based on spectral characteristics and the specific absorption; each class of compounds corresponds to a maximum absorption in the UV spectrum. The phytochemical compounds were identified at specific wavelengths between 700-200 nm. The absorption intensities were measured. The obtained results proved that ethanolic extract showed absorption peaks attributed to: phenolic compounds (free phenolic acids and phenolic acids derivatives) registrated between 220-280 nm, unsymmetrical chemical structure compounds (caffeic acid, chlorogenic acid, ferulic acid) with maximum absorption peak and absorption "shoulder" that may be due to substitution of hydroxyl or methoxy group, flavonoid compounds (in free form or glycosides) between 330-360 nm, due to the double bond in position 2,3 and carbonyl group in position 4 flavonols. UV spectra showed two major peaks of absorption (quercetin glycoside, rutin, etc.). The results obtained by UV-VIS spectroscopy has revealed the presence of phenolic derivatives such as cicoric acid (240 nm), caftaric acid (329 nm), caffeic acid (240 nm), rutoside (205 nm), quercetin (255 nm), luteolin (235 nm) in all three species of Echinacea. The echinacoside is absent. This profile mentioned above and the absence of phenolic compound echinacoside leads to the conclusion that species harvested as Echinacea angustifolia and Echinacea pallida are Echinacea purpurea also; It can be said that preliminary fingerprinting of Echinacea species through correspondence with the phenolic derivatives profile can be achieved by UV-VIS spectroscopic investigation, which is an adequate technique for preliminary identification and authentication of Echinacea in medicinal herbs.

Keywords: Echinacea species, Fingerprinting, Phenolic compounds, UV-VIS spectroscopy

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Authors: S. Mamoucha, N.Tsafantakis, T. Ioannidis, S. Chatzipanagiotou, C. Nikolaou, L. Skaltsounis, N. Fokialakis, N. Christodoulakis

Abstract:

Introduction: Pistacia lentiscus L. (well known as Mastic tree) is an evergreen sclerophyllous shrub that extensively thrives in the eastern Mediterranean area yet only the trees cultivated in the southern region of the Greek island Chios produces mastic resin. Different parts of P. lentiscus L. var. chia have been used in folk medicine for various purposes, such as tonic, aphrodisiac, antiseptic, antihypertensive and management of dental, gastrointestinal, liver, urinary, and respiratory tract disorders. Several studies have focused on the antibacterial activity of its resin (gum) and its essential oil. However, there is no study combining anatomy of the plant organs, phytochemical profile, and antibacterial screening of the plant. In our attempt to discover novel bioactive metabolites from the mastic tree, we screened its antibacterial activity not only against ATCC strains but also against clinical, resistant strains. Materials-methods: Leaves were investigated using Transmission (ΤΕΜ) and Scanning Εlectron Microscopy (SEM). Histochemical tests were performed on fresh and fixed tissue. Extracts prepared from dried, powdered leaves using 3 different solvents (DCM, MeOH and H2O) the waste water obtained after a hydrodistillation process for essential oil production were screened for their phytochemical content and antibacterial activity. Μetabolite profiling of polar and non-polar extracts was recorded by GC-MS and LC-HRMS techniques and analyzed using in-house and commercial libraries. The antibacterial screening was performed against Staphylococcus aureus ATCC25923, Escherichia coli ATCC25922, Pseudomonas aeruginosa ATCC27853 and against clinical, resistant strains Methicillin-resistant S. aureus (MRSA), Carbapenem-Resistant Metallo-β-Lactamase (carbapenemase) P. aeruginosa (VIM), Klebsiella pneumoniae carbapenemases (KPCs) and Acinetobacter baumanii resistant strains. The antibacterial activity was tested by the Kirby Bauer and the Agar Well Diffusion method. The zone of inhibition (ZI) of each extract was measured and compared with those of common antibiotics. Results: Leaf is compact with inosclereids and numerous idioblasts containing a globular, spiny crystal. The major nerves of the leaf contain a resin duct. Mesophyll cells showed accumulation of osmiophillic metabolites. Histochemical treatments defined secondary metabolites in subcellular localization. The phytochemical investigation revealed the presence of a large number of secondary metabolites, belonging to different chemical groups, such as terpenoids, phenolic compounds (mainly myricetin, kaempferol and quercetin glycosides), phenolic, and fatty acids. Among the extracts, the hydrostillation wastewater achieved the best results against most of the bacteria tested. MRSA, VIM and A. baumanii were inhibited. Conclusion: Extracts from plants have recently been of great interest with respect to their antimicrobial activity. Their use emerged from a growing tendency to replace synthetic antimicrobial agents with natural ones. Leaves of P. lentiscus L. var. chia showed a high antimicrobial activity even against drug - resistant bacteria. Future prospects concern the better understanding of mode of action of the antibacterial activity, the isolation of the most bioactive constituents and the clarification if the activity is related to a single compound or to the synergistic effect of several ones.

Keywords: antibacterial screening, leaf anatomy, phytochemical profile, Pistacia lentiscus var. chia

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