Phytochemical Analysis and Anticancer Properties of Drimia maritima Bulb Extracts on Colorectal Cancer Cells.

Cancer is a worldwide health problem and is the second leading cause of death after heart disease. Due to the high cost and severe side effects associated with chemotherapy treatments, natural products with anticancer therapeutic potential may play a promising role in anticancer therapy. The purpose of this study was to investigate the cytotoxic and apoptotic characteristics of the aqueous Drimia maritima bulb extract on Caco-2 and COLO-205 colorectal cancer cells. In order to reach such a purpose, the chemical composition was examined using the GC-MS method, and the selective antiproliferative effect was determined in colon cancer cell lines in normal gingival fibroblasts. The intracellular ROS, mitochondrial membrane potential, and gene expression changes in selected genes (CASP8, TNF-α, and IL-6 genes) were assessed to determine the molecular mechanism of the antitumor effect of the extract. GC-MS results revealed the presence of fifty-seven compounds, and Proscillaridin A was the predominant secondary metabolite in the extract. The IC50 of D. maritima bulb extract on Caco-2, COLO-205, and the normal human gingival fibroblasts were obtained at 0.9 µg/mL, 2.3 µg/mL, and 13.1 µg/mL, respectively. The apoptotic effect assay indicated that the bulb extract induced apoptosis in both colon cancer cell lines. D. maritima bulb extract was only able to induce statistically significant ROS levels in COLO-205 cells in a dose-dependent manner. The mitochondrial membrane potential (MMP) revealed a significant decrease in the MMP of Caco-2 and COLO-205 to various concentrations of the bulb extract. At the molecular level, RT-qPCR was used to assess gene expression of CASP8, TNF-α, and IL-6 genes in Caco-2 and COLO-205 cancer cells. The results showed that the expression of pro-inflammatory genes TNF-α and IL-6 were upregulated. The apoptotic initiator gene CASP8 was also upregulated in the Caco-2 cell line and did not reach significance in COLO-205 cells. These results lead to the conclusion that D. maritima extract induced cell death in both cell lines and may have the potential to be used in CRC therapy in the future.

Beneficial properties of Drimia numidica leaf methanolic extract against the cytogenotoxic effects of mitomycin C on human lymphocytes.

This study investigated the phytochemical profile of Drimia numidica leaf methanolic extract, as well as its cyto-genotoxic and cyto/genoprotective potential against mitomycin C (MMC) mediated effects on healthy human lymphocytes. Photosynthetic pigments, trace elements, and secondary metabolites were estimated and/or identified in methanolic extract of mature leaves, and the latter was further used for assessing its in vitro biological effects on MMC-free and/or MMC-treated human lymphocytes (at low, non-toxic concentrations of 0.001 and 0.01% v/v). The results showed that D. numidica leaf methanolic extract, being rich in carotenoids, phenolics, flavonoids, organic acids and bufadienolides, could be protective against MMC mediated cyto/genotoxic potential in healthy human lymphocytes. Biomolecules possessing antioxidant and antitumor potential, such as beta-carotene and lutein among others, chlorogenic acid, caffeic acid and their derivatives, minerals such as Si, as well as apigenin- and luteolin-derived glycosides, either individual or in a mixture, could be beneficial rather than harmful, at least at the extract concentrations tested. Although further in vitro and in vivo studies are still needed for elucidating the beneficial (individual and/or additive/synergistic) role of those compounds, the results of the present study are quite promising, thus encouraging new challenges for the appropriate utilization of D. numidica leaf extract.

Screening for insecticidal toxicity of Urginea maritima (L.) bulbs and Asphodelus microcarpus (L.) tubers for the control of Tribolium castaneum (Tenebrionidae).

This research was conducted to determine the effect of geographical distribution on morphological characteristics of plants and some chemical compounds such as polyphenols. Moreover, we explore the contact toxicity of ethanolic and methanolic extracts from Urginea maritima bulbs and Asphodelus microcarpus tubers collected from three localities in the North West of Tunisia against Tribolium castaneum adults. The toxicity results demonstrated that both of the studied plant extracts had an effective control against T. castaneum at concentration 7.5 µL/L air, thus total mortality reached. In addition, results showed plant extracts with methanol exhibited high mortality percentage of T. castaneum. On the other hand, results revealed that chemical composition of U. maritime bulbs and A. microcarpus tuber extract depends on the soil components and geographical distribution of plants. The present investigation confirmed that A. microcarpus and U. maritima may be recommended as an eco-friendly alternative to synthetic insecticide against T. castaneum.

Effect οf elicitοrs οn cell suspensiοn culture οf Urginea maritima L. tοwards prοductiοn οf prοscillaridin A.

BACKGROUND: Urginea maritima (L.f.) Baker (Hyacinthaceae) is a perennial bulbous medicinal plant that is currently at risk of extinction. Squill (white sea onion) is an analogous cardiotonic to digitalis. The purpose of the current work was to assess the optimal growth conditions for Urginea cells to synthesize the cardiac glycoside proscillaridin A by involving illumination, carbon source, methyl jasmonate (MJ), and culture system. RESULTS: When cells were cultured for 28 days at 21 ± 2 ∘C in the dark on Murashige and Skoog (MS) media containing 1 mg/L 2, 4-D and 0.5 mg/L Kin and 30 g/L sucrose, the cell proliferation, and proscillaridin A synthesis were effectively controlled. At low concentrations, MJ stimulated the synthesis of proscillaridin A (PsA). High-performance liquid chromatography (HPLC) analysis of suspension extracts demonstrated that the callus maintained in MS media enriched with 1 mg/L 2, 4-D and 0.5 mg/L Kin yielded a greater formation of Proscillaridin A (141.31 mg/g DW) than untreated plants. CONCLUSIONS: The results indicate that in vitro cultures of U. maritima may be an excellent source of proscillaridin A. Moreover, it is one of the most important cardiac glycoside, which has been found to exhibit anticancer activities. Suspension cultures of Urginea cells could be as highly productive as a callus culture.

Phytochemical Screening, Antifungal, and Anticancer Activities of Medicinal Plants Thymelaea Hirsuta, Urginea Maritima, and Plantago Albicans.

Ethyl acetate, ethanol, and acetone extracts of the medicinal plants Thymelaea hirsuta L., Urginea maritima L., and Plantago albicans L. (aerial parts) were evaluated for their phytochemical compositions, antimycotic activity against dermatophytes, and antiproliferative activity against different human cancer cell lines. Among them, the ethanolic extracts showed the highest phytochemical contents along with hyperactivities and were then selected for gas chromatography-mass spectrometry and Fourier-transform infrared spectroscopy analysis. The Fourier-transform infrared spectroscopy analysis confirmed the presence of different characteristic peak values with various functional chemical groups of the active components. However, U. maritima extracts through Fourier-transform infrared spectroscopy analysis showed distinctive peaks related to phenolic, amines, amides, aromatic, alkanes, alkyne, cyclopentanone, conjugated aldehyde, nitro, methoxy, uronic acids, aromatic esters, tertiary alcohol or ester, secondary and primary alcohols, aliphatic ether, sulfoxide, vinylidene, and halo compounds. Many bioactive main compounds with reported biological activities were detected by GC/MS (%) in the ethanolic extract of T. hirsuta, U. maritima, and P. albicans. All studied dermatophytes included a diverse set of virulence factors, including phospholipase, protease, keratinase, hemolysis, and melanoid production, all of which play vital roles in dermatophytic infection. Ethanolic extract of P. albicans inhibited the growth of Trichophyton soudanense totally and Trichophyton erinacei in addition to all Microsporum species. In contrast, the ethanolic extract of Trichophyton hirsuta at concentrations of 25 g/mL totally prevented the growth of all Trichophyton species. EtOH extract of U. maritima completely prevented the growth (100% inhibition) of all dermatophytic strains under study at the lowest concentration of 12.5 µg/mL. Scanning electron microscope analysis revealed considerable morphological modifications and structural alterations in dermatophyte species exposed to ethanolic extract of these plants. The viability of HCT-116, HepG-2, MCF-7, and HeLa cell lines was reduced after treatment with the ethanolic extracts of T. hirsuta, U. maritima, and P. albicans individually with IC50 values (10.0, 9.97, 48.5, and 56.24 µg/mL), (26.98, 25.0, 17.11, and 9.52 µg/mL), and (9.32, 7.46, 12.50, and 16.32 µg/mL), respectively. Our work revealed the significance of these traditional ethnomedical plants as potent sources for biologically active pharmaceuticals with potential applicability for the treatment of fungal and cancer diseases.

Antiproliferative Bufadienolides from the Bulbs of Drimia altissima.

The bulbs of the South African Drimia altissima (Asparagaceae or Hyacinthaceae sensu APGII) have yielded a range of previously undescribed bufadienolides, drimianins A-G (1-7), the known bufadienolides bovogenin A (8), 3ß-O-ß-d-glucopyranosylbovogenin A (9), scillaren F (10), and altoside (11), the known homoisoflavonoid (3S)-3-(4'-methoxybenzyl)-5,6,7-trimethoxychroman-4-one (urgineanin C), the sesquiterpenoids 1ß,6α-dihydroxy-4(15)-eudesmene and 6α-hydroxy-4(15)-eudesmen-1-one, polybotrin, adenosine, and 9R-hydroxy-(10E,12Z)-octadecadienoic acid ethyl ester. The bufadienolides isolated were tested at 10 µM in the NCI-60 cancer cell screen, and nine of these were selected for further screening at five concentrations. Drimianins C (3) and E (5) showed activity at the nanomolar level against a number of human cancer cell lines in the NCI-60 screen.

Isolation and characterisation of altissimin: a novel cytotoxic flavonoid C-apioglucoside from Drimia altissima (Asparagaceae).

Flavonoids are a class of biologically active compounds with various proven nutraceutical benefits. In flavonoid C-glycosides, the aglycones are attached to sugar residues via cleavage-resistant C-C bonds which alter typical flavonoid pharmacokinetic properties. In these compounds, the combination of biological activities from the flavonoid moieties and sugar residues create unique and more diverse biological functions than those of O-glycosylated and unsubstituted flavonoids. Through a series of reverse phase chromatography techniques and various spectroscopic methods, the phytochemical investigation of Drimia altissima (L.F.) Ker Gawl., a specie from the Asparagaceae family, led to the isolation and chemical characterisation of a novel C-glucosylflavonoid, altissimin, with a unique apioglucoside arrangement to the apigenin aglycone. Altissimin was found to possess strong in vitro anti-proliferative activity against HeLa cervical cancer cells.

Therapeutic Aspects of Squill; An Evidence-Based Review.

From ancient times, medicinal plants have been usually utilized to treat many disorders, but today, interest in these herbs is again aroused, because of their fewer side effects and low-cost. In traditional medicine, for many diseases, various medicinal herbs have been suggested so far. Drimia maritime, also named squill, is an important medicinal plant for the treatment of many diseases, especially respiratory diseases. In the current evidence-based study, we conducted a review of the general characteristics, ingredients, administration form, and side effects of squill in traditional medicine. For this purpose, traditional Persian medicine literatures and electronic databases were examined including PubMed, Scopus, and Google Scholar. Many compounds are isolated from D.maritima, including scillaren, scillirubroside, scillarenin, and bufadienolide glycosides. Oxymel is the most commonly used form of squill for various diseases, especially respiratory diseases. Besides, squill has been used in the treatment of cardiovascular, digestive, and dermatological disorders, it is also used against various cancer cells for its antioxidant and cytotoxic properties. Moreover, there is relatively reliable evidence of its benefits for bacterial and helminthic infections, rheumatism, edema, gout, abortion induction, healing of wounds and urine induction. It seems that supplementary studies are required to explore the bioactive agents and their effective mechanisms.

Drimia indica: A Plant Used in Traditional Medicine and Its Potential for Clinical Uses.

Drimia indica (Roxb.) Jessop (Asparagaceae) is a reputed Ayurvedic medicine for a number of therapeutic benefits, including for cardiac diseases, indigestion, asthma, dropsy, rheumatism, leprosy, and skin ailments. The present work aimed to critically and extensively review its traditional uses, phytochemistry, pharmacology, toxicology, and taxonomy together with the mechanisms of action of selected extracts of D. indica. A systematic literature survey from scientific databases such as PubMed, Scopus, and Web of Science as well as from some textbooks and classical texts was conducted. The plant, mainly its bulb, contains various bioactive constituents, such as alkylresorcinols, bufadienolides, phytosterols, and flavonoids. Various scientific studies have proven that the plant has anthelmintic, anticancer, antidiabetic, antimicrobial, antioxidant, and wound healing activities. The present work concludes that D. indica has the potential to treat various diseases, mainly microbial infections. This review also suggests that bufadienolides, flavonoids, and steroids might be responsible for its bioactive potential.

C-glycosyl flavone from Urginea indica inhibits proliferation & angiogenesis & induces apoptosis via cyclin-dependent kinase 6 in human breast, hepatic & colon cancer cell lines.

BACKGROUND & OBJECTIVES: Search for novel compounds beneficial to the treatment of cancer attracts a great deal of attention. We earlier demonstrated the isolation of 5,7-dihydroxy-2-[4'-hydroxy-3'-(methoxymethyl)phenyl]-6-C-ß-glucopyranosyl flavone, a novel C-glycosyl flavone from Urginea indica bulb. The present study was undertaken to investigate the effect of this novel compound on human normal epithelial and breast, hepatic and colon cancer cell lines. METHODS: : The maximum non-toxic concentration (MNTC) and cytotoxicity of C-glycosyl flavone were assayed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Cell cycle was analyzed by flow cytometry. Docking studies were performed to predict possible targets. Levels of cyclin-dependent kinase 1 (CDK1) and CDK6, Bcl2 and BAX and cytochrome c were quantified by specific ELISA. Mitochondrial membrane potential was determined using JC-1 dye. Apoptosis was quantified by Annexin V ELISA method. RESULTS: : Flow cytometry analysis demonstrated G0/G1 arrest. In silico docking studies predicted CDK1 and CDK6 as a possible target of C-glycosyl flavone. In vitro study confirmed CDK6 as the main target in C-glycosyl flavone-treated cancer cell lines. C-glycosyl flavone treatment also induced membrane blebbing, chromatin fragmentation and nucleosome formation. C-glycosyl flavone treatment caused marked loss of mitochondrial membrane potential, decrease in Bcl2/BAX ratio and activation of caspase-3 and release of caspase-9 and cytochrome c. In addition, C-glycosyl flavone inhibited the tumour-induced angiogenesis and reduced the vascular endothelial growth factor levels. Similarly, CDK6 inhibitor significantly inhibited proliferation and angiogenesis and induced apoptosis in tested cell lines. INTERPRETATION & CONCLUSIONS: The results indicate that C-glycosyl flavone may exert induction of apoptosis, cell cycle arrest and inhibition of angiogenesis via CDK6. Thus, targeting CDK6 using C-glycosyl flavone may serve as a novel therapeutic approach for the treatment of breast, hepatic and colon cancers.

C-glycosyl Flavone from Urginea indica Inhibits Growth and Dissemination of Ehrlich Ascites Carcinoma Cells in Mice.

BACKGROUND: C-glycosyl flavone, a phytochemical constituent in U.indica bulb, has been reported to possess cytotoxic activity. OBJECTIVE: The present study aims to investigate the toxicity and anticancer potentials of C-glycosyl flavone against Ehrlich ascites carcinoma mice model. METHOD: In present study, acute and chronic toxicity along with antitumor activity of C-glycosyl flavone isolated from U.indica bulb were Performed using in vitro and in vivo methods. Acute and chronic toxicity of C-glycosyl flavone was evaluated using Swiss albino mice. The effect of C-glycosyl flavone on proliferation of Ehrlich ascites carcinoma (EAC) cells was determined. Further, growth inhibition and dissemination were studied using EAC induced mice model. RESULTS: C-glycosyl flavone showed significant therapeutic potency against EAC cells in terms of reduced viability, cell cycle arrest, induction of apoptosis, inhibition of capillary formation, reduced VEGF levels. Moreover, there was reduction in body weight, tumor volume, viable tumor cells, increased survival of EAC induced mice upon C-glycosyl flavone treatment. Treatment also reduced dissemination of EAC cells into heart, kidney, liver and brain and diminished the pathological alterations induced by EAC cells in mice. In addition, there was an improvement in hemoglobin levels and counts of RBC, neutrophils, lymphocytes and monocytes in C-glycosyl flavone-treated mice with tumor. An enhancement of antioxidant status in C-glycosyl flavone treated EAC-bearing mice which appeared in terms of decreased serum thiobarbituric acid reactive substance and lipid peroxidation, increased GSH, SOD, Catalase and GPX. These results were comparable to a standard 5- fluorouracil treatment. C-glycosyl flavone exhibited safety profile in toxicity studies. CONCLUSION: Our study confirms the therapeutic potency of C-glycosyl flavone against EAC in inhibition of dissemination and growth of EAC in mice.

Traditional medical uses of Drimia species in terms of phytochemistry, pharmacology and toxicology.

Drimia genus includes plants that used from ancient time for various ailments such as dropsy, respiratory ailment, bone and joint complications, skin disorders, epilepsy and cancer. Toxic properties of some Drimia species also were noted by ancient scientists and these plants have been traditionally used for rat control. Bufadienolides have been identified as the main constituents in the genus of Drimia. Phenolics, sterols, protein and some of other phytochemicals have been also isolated from these plants. Pharmacological and clinical studies have strongly approved their effect on cardiovascular system. Extracts and compounds isolated from Drimia species showed biological activities such as antibacterial, antifungal, antiviral, antioxidant, anti-inflammatory and insecticidal effects through several in vivo and in vitro studies. Moreover, cytotoxic and antitumor activities which may be related to bufadienolide content of these plants have been considered by many researchers. Traditional therapeutic values of these plants for treating respiratory and rheumatic ailments as well as skin disorders are needed to be validated through more researches. Toxic effects of these plants and isolated compounds have been investigated through several in vivo studies. Drimia plants and their isolated compounds have narrow therapeutic index, so patients should be prohibited from applying these plants without medical supervision and should be informed about the main intoxication symptoms before starting treatment. Moreover, interaction of Drimia plants with other constituents of traditional herbal mixtures as well as chemical and biological modalities for reducing toxicity of bufadienolide compounds can be subjected for future studies.

Treatment of Recurrent Ovarian Cysts and Primary Infertility by Iranian Traditional Medicine: A Case Report.

Infertility is a medical and psychosocial problem with a high prevalence. There are different treatments for this problem in Iranian traditional medicine. A 28-year-old woman presented with the complaints of 4 emergency operations of the left ovarian cyst during 4 years and infertility. Diagnostic laparoscopy showed an ovarian cyst, adhesion, and endometriosis. Hysteroscopy was unremarkable. After 2 months of letrozole administration, the ovarian cyst ruptured again. Considering the failure of conventional treatments, Iranian traditional medicine products were administered to the patient. After 3 months, the patient conceived and delivered a healthy boy through normal vaginal delivery. These compounds may help with pregnancy as a uterine tonic, vitalizer, and aphrodisiac with brain and cardiac tonic properties.

Antiproliferative Homoisoflavonoids and Bufatrienolides from Urginea depressa.

Investigation of the South African plant Urginea depressa Baker (Asparagaceae Juss.) for antiproliferative activity against the A2780 ovarian cancer cell line led to the isolation of the six new homoisoflavonoids urgineanins A-F (1-6), the two known bufatrienolides 7 and 9, and the new bufatrienolides urginins B and C (8 and 10). Structures were elucidated based on analysis of their 1D and 2D NMR spectra, electronic circular dichroism, and mass spectrometric data. Five of the six new homoisoflavonoids had good antiproliferative activity against the A2780 ovarian cancer, A2058 melanoma, and H522-T1 human non-small-cell lung cancer cells, and urgineanin A (1) had submicromolar activity against all three cell lines. The four bufatrienolides 7-10 had strong antiproliferative activity against the same cell line, with IC50 values of 24.1, 11.2, 111, and 40.6 nM, respectively.

The traditional medical uses and cytotoxic activities of sixty-one Egyptian plants: discovery of an active cardiac glycoside from Urginea maritima.

ETHNOPHARMACOLOGICAL RELEVANCE: Medicinal plants from the Sinai desert are widely used in traditional Bedouin medicine to treat a range of conditions including, cancers, and may thus be useful sources of novel anti-tumor compounds. Information on plants used in this way was obtained through collaboration with Bedouin herbalists. AIM OF THE STUDY: To document the traditional uses of 61 species from 29 families of Egyptian medicinal plants and to investigate their biological activity using a cytotoxicity assay. MATERIAL AND METHODS: MeOH extracts of the 61 plant species investigated were dissolved in 10% DMSO and their cytotoxic activity was evaluated. The extracts were tested in duplicate on three separate occasions at three different concentrations (1, 10 and 100µg/ml) against human lymphoma U-937 GTB. The most active extract was subjected to bioassay-guided fractionation using HPLC and LC/ESI-MS to isolate and identify its active components. RESULTS AND DISCUSSION: The most potent extracts were those from Asclepias sinaica, Urginea maritima, Nerium oleander and Catharanthus roseus, followed by those from Cichorium endivia, Pulicaria undulate and Melia azedarach. Literature reports indicate that several of these plants produce cardiac glycosides. Bioassay-guided fractionation of alcoholic U. maritima extracts led to the isolation of a bioactive bufadienolide that was subsequently shown to be proscillaridin A, as determined by 1D and 2D NMR spectroscopy. This result demonstrates the value of plants used in traditional medicine as sources of medicinally interesting cytotoxic compounds.

Major antifungal activity from the bulbs of Indian squill Urginea indica is a chitinase.

We have identified a chitinase with antifungal activity in the bulbs of the plant Urginea indica(Indian squill) and purified it about 26-fold. The purified preparation contained a Mr 29 kDa protein that was an active growth inhibitor of the fungal pathogens Fusarium oxysporum and Rhizoctonia solani in an in vitro assay. Amino acid sequence analysis of the Mr 29 kDa protein revealed it to be highly homologous to the family 19 glycoside hydrolases, which are known to possess chitinase activity. The U. indica chitinase lacked a cysteine-rich N-terminal domain (characteristic of class I chitinases) and contained a conserved motif indicative of the signature 1 of family 19 glycoside hydrolases. It shared a approximately 70% sequence identity with the 26 kDa endochitinase of Hordeum vulgare, a typical class II chitinase of family 19. The five cysteines in the partial sequence of the Mr 29 kDa chitinase were found to be identical in location to five of the seven cysteines present in the catalytic domain of the H. vulgare enzyme. The molecular weight, the lack of an N-terminal cysteine-rich sequence, and the striking identity to the H. vulgare endochitinase suggest that the Mr 29 kDa U. indica protein is a putative class II chitinase. The antifungal activity is presumably mediated through the chitinolytic activity of the Mr 29 kDa protein.

Antiangiogenic and proapoptotic activity of a novel glycoprotein from U. indica is mediated by NF-kappaB and Caspase activated DNase in ascites tumor model.

We have identified a novel glycoprotein from Urginea indica bulbs with potent in vivo antitumor activity against growth of an ascites tumor, mouse mammary carcinoma. In this paper we report characterization of a 29 kDa glycoprotein from U. indica and demonstrate the mechanism of antiangiogenic and proapoptotic activity. N-terminal sequence of the high performance liquid chromatography (HPLC) pure glycoprotein showed sequence homology to an extent of 40-50% with known angiogenesis inhibitor and apoptosis-inducing protein from C. elegans and G. gallus respectively. Our results on antiangiogenic property of the glycoprotein include inhibition of in vivo angiogenesis assays, decreased micro vessel density count and CD31 antigen staining in 29 kDa glycoprotein treated mice peritoneum. In vitro inhibition of vascular endothelial growth factor induced proliferation of human umbilical vein endothelial cells (HUVECs) by the glycoprotein further supports its antiangiogenic activity. The mechanism of antiangiogenesis involved inhibition of translocation of nuclear factor kappa B to the nucleus resulting in decreased expression of vascular endothelial growth factor gene as is demonstrated by our results on quantification of vascular endothelial growth factor levels in the glycoprotein treated tumor bearing mice. Our results on activation of Caspase-3 with concomitant translocation of caspase activated DNase to the tumor cell nuclei resulting in DNA fragmentation induced by the glycoprotein in vivo clearly demonstrated a parallel proapoptotic activity of the glycoprotein.