Transient receptor potential channel stimulation induced oxidative stress and apoptosis in the colon of mice with colitis-associated colon cancer: modulator role of Sambucus ebulus L.

BACKGROUND: Increased Ca2+ entry causes an increase in tumor cell proliferation, apoptosis, cytosolic reactive free oxygen species (cyROS), and mitochondrial ROS (miROS) in tumor cells. The cyROS and miROS stimulate the cation channels, including the TRPA1, TRPM2, and TRPV1. Sambucus ebulus L (SEB) (Dwarf Elder) induced both antioxidant and anticancer effects in the human hepatocarcinoma and human colon carcinoma cancer cell lines. We investigated the etiology of colorectal cancer and the impact of three channels, as well as the protective effects of SEB on apoptosis, cyROS, and miROS in the colon of mice with colitis-associated colon cancer (AOM/DSS). METHODS: A total 28 mice were equally divided into four groups as control, SEB (100 mg/kg/day for 14 days), AOM/DSS, and SEB + AOM/DSS. Azoxymethane/dextran sulfate sodium-induced colon cancer associated with colitis was induced in the AOM/DSS groups within 10 weeks. At the end of the experiments, the colon samples were removed from the mice. RESULTS: The protein bands of caspase - 3, TRPA1, TRPM2, and TRPV1 were increased by the treatments of AOM/DSS. The levels of apoptosis, cyROS, cleaved caspase - 3, and cleaved caspase - 9, as well as the depolarization of the mitochondrial membrane, all increased in the AOM/DSS group. Although they were reduced in the SEB and AOM/DSS + SEB groups by the treatments of SEB, TRPA1 (AP18), TRPM2 (ACA), and TRPV1 (capsazepine) antagonists, the apoptotic and oxidant values were further elevated in the AOM/DSS group by the treatments of TRPA1 (cinnamaldehyde), TRPM2 (H2O2), and TRPV1 (capsaicin) agonists. CONCLUSION: The activations of TRPA1, TRPM2, and TRPV1 channels induced the increase of apoptotic and oxidant actions in the colon cancer cells, although their inhibition via SEB treatment decreased the actions. Hence, TRPA1, TRPM2, and TRPV1 activations could be used as effective agents in the treatment of colon tumors.

A novel sensing platform for electrochemical detection of metronidazole antibiotic based on green-synthesized magnetic Fe3O4 nanoparticles.

The spread of antibiotic resistant genes has become a serious global concern. Thus, the development of efficient antibiotic monitoring systems to reduce their environmental risks is of great importance. Here, a potent electrochemical sensor was fabricated to detect metronidazole (MNZ) on the basis of green synthesis of Fe3O4 nanoparticles (NPs) using Sambucus ebulus L. leaves alcoholic plant extract as a safe and impressive reducing and stabilizing agent. Several analyses such as X-ray diffraction (XRD), Fourier transform infrared spectrophotometer (FTIR), thermogravimetric analysis (TGA), field emission scanning electron microscope (FESEM), energy dispersive X-ray spectroscopy (EDX), and dynamic light scattering (DLS) confirmed the production of homogeneous, monodisperse, regular, and stable magnetite NPs with a spherical morphology. The as-prepared Fe3O4NPs were afterwards applied to evaluate the electrochemical activity of MNZ by merging them with graphene nanosheets (GR NSs) on the glassy carbon electrode (GCE). The GR/Fe3O4NPs/GCE represented extraordinary catalytic activity toward MNZ with two dynamic ranges of 0.05-5 µM and 5-120 µM, limit of detection (LOD) of 0.23 nM, limit of quantification (LOQ) of 0.76 nM, and sensitivity of 7.34 µA µM-1 cm-2. The fabricated sensor was further employed as a practical tool for electrochemical detection of MNZ in real aqueous samples.

Inhibition of herpes simplex virus type 1 infection by Sambucus ebulus extract in vitro.

Background: The emergence of drug-resistant strains of herpes simplex virus type 1 (HSV-1) has been increasingly reported. Therefore, attempts to discover new antiviral agents in particular from natural compounds are required. In this study, we evaluated the possible inhibitory effects of hydroalcoholic extract of Sambucus ebulus (S. ebulus ) against HSV-1. Methods: S. ebulus extract was produced by maceration method. MTT assay was used to evaluate the cytotoxicity effects of the S. ebulus extract; also, antiviral effects were measured both by test TCID50 and quantitative real-time PCR methods. To study the inhibitory impact of S. ebulus extract on the expression of HSV-1 antigens, indirect immunofluorescence assay (IFA) was also performed. All analyses were performed using the GraphPad Prism software v. 7.0. Results: In the postexposure assay of HSV-1 with S. ebulus extract at the highest nontoxic concentration (75 µg/mL), S. ebulus extract led to 2.6 log10 TCID50 reduction in infectious virus titer. At the highest nontoxic concentration, the S. ebulus extract led to inhibition rates of 91.2%, based on the quantitative real-time PCR assay results (p<0.001). Also, in the immunofluorescence assay, a significant reduction was observed in fluorescence emission intensity in HSV-1-infected cell treated with S. ebulus extract compared to the control group. Conclusion: S. ebulus extract is a novel and effective natural compound in reducing HSV-1 titer and future studies should be conducted to discover the complete mechanism of antiviral effect of this natural compound.

Sambucus ebulus extract stimulates cellular responses in cutaneous leishmaniasis.

This is the first study aiming to determine the therapeutic effects of the Sambucus ebulus aquatic extract as an antileishmanial herbal drug and evaluate the immune responses in Leishmania major major infected BALB/c mice. The antileishmanial activity of S ebulus aquatic extract was evaluated using MTT test as well as parasite rescue and transformation assay. Footpad swelling and parasite load of infected mice were measured by several techniques. The immune responses were evaluated by measuring the levels of IFN-γ, IL-4, nitric oxide and arginase. The results indicated that S. ebulus can significantly decrease L. major promastigotes and amastigotes viability, but it was not toxic to macrophages. The lesion size, parasite burden and the level of ARG decreased in the treated infected mice, while the IFN-γ-to-IL-4 ratio and the level of NO increased significantly. Altogether, the S. ebulus extract is an effective compound for killing Leishmania parasite without excessive toxicity to the host cells and can cure the CL by switching the host immune responses towards Th1 response. Thus, it may be a perfect therapeutic option for CL treatment.

Ebulin-RP, a novel member of the Ebulin gene family with low cytotoxicity as a result of deficient sugar binding domains.

BACKGROUND: Sambucus ebulus is a rich source of ribosome-inactivating proteins (RIPs) and RIP-related lectins generated from multiple genes. These proteins differ in their structure, enzymatic activity and sugar binding specificity. METHODS: We have purified and characterized ebulin-RP from S. ebulus leaves and determined the amino acid sequence by cDNA cloning. Cytotoxicity was studied in a variety of cancer cells and a comparative study of the ability of ebulin-RP to bind sugars using "in vitro" and "in silico" approaches was performed. RESULTS: Ebulin-RP is a novel heterodimeric type 2 RIP present in S. ebulus leaves together with the type 2 RIP ebulin l, which displayed rRNA N-glycosidase activity but unlike ebulin l, lacked functional sugar binding domains. As a consequence of changes in its B-chain, ebulin-RP displayed lower cytotoxicity than ebulin l towards cancer cells and induced apoptosis as the predominant pattern of cell death. CONCLUSIONS: Ebulin-RP is a novel member of the ebulin gene family with low cytotoxicity as a result of deficient sugar binding domains. Type 2 RIP genes from Sambucus have evolved to render proteins with different sugar affinities that may be related to different biological activities and could result in an advantage for the plant. GENERAL SIGNIFICANCE: The ebulin family of RIPs and lectins can serve as a good model for studying the evolutionary process which may have occurred in RIPs. The lack of cytotoxicity of ebulin-RP makes it a good candidate as a toxic moiety in the construction of immunotoxins and conjugates directed against specific targets.

Investigation of the effect of enzymatic and alkali treatments on the physico-chemical properties of Sambucus ebulus L. plant fiber.

The investigation aims to determine the effect of enzymatic and alkali treatments on Sambucus ebulus L. stem fiber. For this purpose, Sambucus ebulus L. stem fibers were treated with alkali, cellulase, and pectinase enzymes. An image processing technique was developed and implemented to calculate the average thicknesses of Sambucus ebulus L. fibers. The thickness of alkali, cellulase and pectinase enzyme treated fibers was determined as 478.62 µm, 808.28 µm and 478.20 µm, respectively. Scanning electron microscopy analysis illustrated that enzymatic and alkali treatments lead to the breakage of fiber structure. Furthermore, enzymatic and alkali treatments induce variations in elemental ingredients. All treatments increased the crystallinity index of Sambucus ebulus L. fiber from 72 % (raw fiber) to 83 % (alkali treated), 75.2 % (cellulase enzyme treated) and 86.3 % (pectinase enzyme treated) due to the hydrolysis of hemicellulose. Fourier transform infrared analysis indicated that there are no significant differences in functional groups. Thermogravimetric analysis shows that enzymatic and alkali treatments improve final degradation temperature of the fiber. Mechanical behaviors of cellulase enzyme-treated fiber decrease compared to raw fiber, while pectinase enzyme and alkali treatment cause to improve mechanical properties. Tensile strength of samples was determined as 76.4 MPa (cellulase enzyme treated fiber), 210 MPa (pectinase enzyme treated fiber) and 240 MPa (alkali treated fiber). Young's modules of cellulase enzyme, pectinase enzyme and alkali treated fibers were predicted as 5.5 GPa, 13.1 GPa and 16.6 GPa. Elongation at break of samples was calculated as 5.5 % (cellulase enzyme treated fiber), 6.5 % (pectinase enzyme treated fiber) and 6 % (alkali treated fiber). The results suggest that enzymatic and alkali treatments can modify the functional and structural attributes of Sambucus ebulus L. fiber.

In Vitro and In Vivo Anti-parasitic Activity of Sambucus ebulus and Feijoa sellowiana Extracts Silver Nanoparticles on Toxoplasma gondii Tachyzoites.

BACKGROUND: Current chemical treatments for toxoplasmosis have side effects, researchers are looking for herbal remedies with minimal side effects and the best effectiveness. This study aimed to evaluate the anti-toxoplasmic effects of silver nanoparticles based on Sambucus ebulus (Ag-NPs-S. ebulus) and Feijoa sellowiana (Ag-NPs-F. sellowiana) fruit extracts, in vitro and in vivo. METHODS: Vero cells were treated with different concentrations (0.5, 1, 2, 5, 10, 20, 40 µg/mL) of extracts and pyrimethamine as a positive control. Vero cells were infected with T. gondii and treated with extracts. The infection index and intracellular proliferation of T. gondii were evaluated. The survival rate of infected mice with tachyzoites of T. gondii was examined after intraperitoneal injection of the extracts at a dose of 40 mg/kg/day for 5 days after infection. RESULTS: The Ag-NPs-S. ebulus and Ag-NPs-F. sellowiana, almost similar to pyrimethamine, reduced proliferation index when compared to untreated group. Also, high toxoplasmicidal activity was observed with Ag-NPs-S. ebulus extract. Mice in the treatment groups of Ag-NPs-S. ebulus and pyrimethamine achieved better results in terms of survival than the others. CONCLUSION: The results indicated that Ag-NPs-F. sellowiana and S. ebulus have a significant growth effect on T. gondii in vitro and in vivo. Ag-NPs-S. ebulus extract has a more lethal effect on the parasite than Ag-NPs-F. sellowiana. It is suggested that in future investigate the induction of Toxoplasma-infected cell apoptosis using nanoparticles.

High resolution LC-MS/MS characterization of polyphenolic composition and evaluation of antioxidant activity of Sambucus ebulus fruit tea traditionally used in Bulgaria as a functional food.

The purpose of this study was to perform phytochemical analysis of tea from Sambucus ebulus fruits concerning hydroxycinnamic acids, flavonol glucosides, stilbenes and proanthocyanidin mono-, di- and trimers content. In total, 33 compounds were identified and quantified using UPLC-DAD-ESI/MS/MS system and the results are presented in mg/g dry weight (DW). Among analyzed hydroxycinnamic acids, 5-Caffeoylquinic acid (114.17 mg/g) was most abundant, followed by 3-p-Coumaroylquinic acid (50.33 mg/g) and 3-p-Feruloylquinic acid, p-Coumaric acid glucoside and 4-p-Coumaroylquinic acid (31.36 mg/g, 29.78 mg/g and 27.70 mg/g, respectively). Flavonol glucosides were represented predominantly by Quercetin-3-O-galactoside, Quercetin-3-O-rhamnosyl-galactoside Quercetin-3-O-glucoside and Quercetin-3-O-rhamnosyl-glucoside (3.68 mg/g, 3.22 mg/g, 2.87 mg/g and 2.56 mg/g, respectively). trans-Resveratrol-3-O-glucoside, epicatechin (40.62 mg/g) and proanthocyanidin di- and -trimers (19.90 mg/g - 31.42 mg/g) also were present in the tea. ABTS cation decolorization assay revealed 1.248 mM UAE activity and the percent of DPPH radical scavenging was 14.25%, corresponding to 39.07 µM Trolox equivalents.

Effects of Sambucus ebulus Extract on Cell Proliferation and Viability of Triple- Negative Breast Cancer: An In Vitro and In Vivo Study.

BACKGROUND: Triple-negative breast cancer (TNBC) accounts for 15% of all breast cancer (BC) cases and is a severe type of BC. Since medicinal herbs containing biocompatible substances that are accepted by patient more than chemical therapeutics, they can be considered a safe option for treating BC. OBJECTIVE: This study evaluated the effect of Sambucus Ebulus (S. ebulus) extract on a model of TNBC. METHODS: S. ebulus extract was prepared using petroleum ether, ethyl acetate, and methanol. The petroleum ether extract was fractionated and analyzed using vacuum liquid chromatography and GC-MS, respectively. MDAMB- 231 and MCF-10A were used as TNBC and normal breast cells, respectively. Flowcytometry and MTT assays were performed to evaluate cell cycle, apoptosis, and viability of the cells. Gene expression analysis was performed using RT-qPCR. Nude mouse allograft tumor models were used, and pathological sections were evaluated. RESULTS: The findings indicated that S. ebulus extract remarkably decreased cell proliferation and viability. The extract had no toxicity to the normal breast cells but efficiently killed the cancer cells. Cell cycle- and apoptosisrelated gene expression showed that fraction 4 of S. ebulus extract significantly increased the expression of Bax, Bak, P53, and c-MYC. CONCLUSION: This study showed satisfactory results of the effect of S. ebulus extract on clearing BC cells both in vitro and in vivo. Thus, S. ebulus extract may be a safe herbal compound for eliminating BC cells without toxicity to host cells.

The amelioration of nicotine-induced reproductive impairment in male mouse by Sambucus ebulus L. fruit extract.

Nicotine as a toxic agent in cigarette smoke impairs the reproductive system. Sambucus ebulus extract (SEE) is shown to have some beneficial effects such as antioxidant properties. The aim of this study was to evaluate the effects of SEE on the hormones of the pituitary-gonadal axis, lipid peroxidation index, antioxidant enzymes, spermatogenesis, and epididymal sperm parameters in male mice treated with nicotine. Adult male mice were divided into five groups; A: normal saline, B: 1 mg/kg nicotine, C: 1 mg/kg nicotine and 10 mg/kg SEE, D: 1 mg/kg nicotine and 50 mg/kg SEE, D: 1 mg/kg nicotine and 100 mg/kg SEE. Treatments lasted for 35 days. The spermicidal activity of SEE was tested in vitro. Sperm count, motility and morphology were assessed for fertility. Serum testosterone, prolactin and luteinizing hormone (LH) were measured, using ELISA. Serum malondialdehyde (MDA), catalase (CAT), and superoxide dismutase (SOD) activity were measured, using colorimetric assays. Spermatogenesis was evaluated by Johnsen's score and morphometry in histological slides. SEE at different doses did not have any spermicidal activity. Sperm parameters were reduced in the nicotine-treated group, compared with controls (P<0.01). Nicotine reduced testosterone and LH levels (P<0.01) and increased prolactin (P<0.01). A hike in MDA and a reduction in SOD activity without change on CAT, were observed in the nicotine group. Nicotine caused hypospermatogenesis. SEE improved most of the above-mentioned parameters, especially in the doses of 50 and 100 mg/kg. Beneficial effects of SEE in the doses of 50 and 100 mg/kg on male reproduction impairment, induced by nicotine might be partly attributed to the reduction of oxidative stress and changes in the hormones of the pituitary-gonadal axis.

Comparison of the Effects of Sambucus ebulus Leaf and Fruit Extracts on Leishmania major In Vitro.

BACKGROUND: Leishmaniasis is one of the major diseases caused by the intracellular parasite of Leishmania. It has become one of the most dangerous health problems today. Our aim of the present study is to compare the effects of Sambucus ebulus leaf and fruit extracts on Leishmania major in vitro. METHODS: In this study, we used MTT, promastigote and amastigote assay to evaluate the effect of different concentrations of the extract on parasite and we compared their effects. The flow cytometry technique was also used to detect the apoptotic effect of the extracts on promastigotes. RESULTS: According to MTT experiment IC50 concentration of leaf and fruit extracts on parasite was 157 µg/ml and 265 µg/ml, respectively. After analysis by flow cytometry, leaf and fruit extracts also showed the apoptosis effect. Leaf and fruit extract caused 40.2 and 2.67 percent apoptosis. CONCLUSION: Based on the above assessment, we determined that the S. ebulus leaf extract has a more toxic effect on promastigotes and amstigotes than its fruit extract and maybe in the future that be used as a drug candidate.

Phytochemical Composition, Anti-Inflammatory and ER Stress-Reducing Potential of Sambucus ebulus L. Fruit Extract.

Sambucus ebulus L. (SE) fruits are used for their immunostimulation, hematopoietic and antiviral potential. Recently, we focused on analyzing the mechanism underlying SE fruit aqueous extract's (FAE) immunomodulation and anti-inflammatory activities, with attention to its endoplasmic reticulum (ER) stress-reducing potential. J774A.1 macrophages were treated with SE FAE alone or in conditions of lipopolysaccharides (LPS) stimulation. Using GC-MS and LC-MS/MS, its phytochemical composition was analyzed. To measure transcription and protein levels, we used qPCR and Western blot, respectively. The prevailing phytochemicals in SE FAE were hydroxycinnamic acids, proanthocyanidins and anthocyanins. The content of some amino acids, organic acids, alcohols, fatty acids and esters were newly reported. Extracts exerted an immunostimulation potential by stimulating IL-6, TNFα, Ccl2, COX2 and iNOS transcription, without inducing ER stress. SE FAE suppressed the LPS-induced transcription of inflammation related genes (IL-1ß, IL-6, TNFα, Ccl2, Icam-1, Fabp4, COX2, iNOS, Noxo1, IL-1ra, Sirt-1) and reduced the protein levels of iNOS, peIF2α, ATF6α and CHOP. The effects were comparable to that of salicylic acid. SE suppresses LPS-stimulated inflammatory markers on the transcription and translation levels. Targeting ER stress is possibly another mechanism underlying its anti-inflammatory potential. These findings reveal the potential of SE fruits as a beneficial therapeutic of inflammation and ER stress-related pathological conditions.

A Comparison of the Effect of Topical Preparation of Sambucus ebulus L. and Hydrocortisone on Hand Eczema: A Double-Blind Randomized Controlled Trial.

Objectives: Corticosteroids as the main treatment of hand eczema can cause major side effects. This study compared the effect of topical preparation of Sambucus ebulus L. leaves and hydrocortisone on the severity of hand eczema. Design: Ninety-four patients with hand eczema aging 18-60 years were recruited in two groups (S. ebulus vs. hydrocortisone). Interventions: The patients used topical medications twice a day and were followed for 4 weeks. The observations were made at the first visit, and also second and fourth weeks of the study. Outcome measures: The primary outcomes were changes in the severity of hand eczema (hand eczema severity index [HECSI]) and life quality (dermatology life quality index [DLQI]). Secondary outcomes were the rate of healing and the severity of itching. Results: Thirty-eight patients in hydrocortisone and 43 in S. ebulus fulfilled all phases of the follow-up. In both groups, the HECSI, itching scores decreased over time without any significant difference between the groups (p = 0.49, 0.57, respectively). The DLQI scores were significantly better in the S. ebulus group (p = 0.02) after 4 weeks of medication. There was no significant difference between the healing rates of the two groups (p = 0.33). Conclusions: Topical use of S. ebulus can be as effective as that of hydrocortisone when it comes to reducing the severity of hand eczema.

A new iridoid diglycoside from Sambucus ebulus L.

The phytochemical examination of the polar constituents of Sambucus ebulus L. leaves led to the identification of patrinoside (1) and of a new diglycoside iridoid, patrinoside-aglycone-11-O-[ß-D-glucopyranosyl-(1→6)-2'-deoxy-ß-D-glucopyranoside] (trivially named as sambuloside) (2). Both of these structures have been assigned by spectroscopic means (NMR and MS).

Unexpected Toxicity of Green Tea Polyphenols in Combination with the Sambucus RIL Ebulin.

The safety of concentrated food complements intake is a major health concern. It has been well established that green tea polyphenols (GTPs) consumption promotes healthy effects. However, the ingestion of large amounts of GTPs is a matter of controversy due to reported adverse effects. We underwent a preliminary exploration of the effects of the oral administration of a standardized concentrated GTPs preparation on mice which suffered from reversible intestinal derangement promoted by sublethal amounts of the antiribosomal lectin ebulin f from dwarf elder (Sambucus ebulus L.). Neither independent oral administration of 30 mg/kg body weight Polyphenon 60 nor intraperitoneal administration of 2.5 mg/kg body weight ebulin f triggered lethal toxicity. In contrast, the simultaneous administration of these same doses of both Polyphenon 60 and ebulin f triggered an important and unexpected synergistic toxic action featured by the biphasic reduction of weight, which continued after eight days, reaching a reduction of 40%. Lethality appeared 2 days after the onset of the combined treatment and reached more than 50% after 10 days.

Polyphenolic Extract from Sambucus ebulus L. Leaves Free and Loaded into Lipid Vesicles.

The paper deals with the preparation and characterisation of hydroalcoholic polyphenolic extract from Sambucus ebulus (SE) leaves that was further loaded into three-types of lipid vesicles: liposomes, transfersomes, and ethosomes, to improve its bioavailability and achieve an optimum pharmacological effect. For Sambucus ebulus L.-loaded lipid vesicles, the entrapment efficiency, particle size, polydispersity index and stability were determined. All prepared lipid vesicles showed a good entrapment efficiency, in the range of 75-85%, nanometric size, low polydispersity indexes, and good stability over three months at 4 °C. The in vitro polyphenols released from lipid vehicles demonstrated slower kinetics when compared to the free extract dissolution in phosphate buffer solution at pH 7.4. Either free SE extract or SE extract loaded into lipid vesicles demonstrated a cytoprotective effect, even at low concentration, 5 ug/mL, against hydrogen peroxide-induced toxicity on L-929 mouse fibroblasts' cell lines. However, the cytoprotective effect depended on the time of the cells pre-treatment with SE extract before exposure to a hydrogen peroxide solution of 50 mM concentration, requiring at least 12 h of pre-treatment with polyphenols with radical scavenging capacity.

In vitro antifungal activity of some traditional Persian medicinal plants on pathogenic fungi.

OBJECTIVE: To investigate the antifungal activities of the extracts and sub-fractions of Phlomis olivieri, Verbascum speciosum, Sambucus ebulus and Erigeron hyrcanicus, four Persian medicinal plants used in Iranian folk medicine. METHODS: Evaluation of the antifungal activity was performed on the clinical isolates of pathogenic fungi including Aspergillus fumigatus, A. flavus, Trichophyton mentagrophytes, T. rubrum, T. verrucosum, Microsporum canis, M. gypseum and Epidermophyton floccosum, and the yeast Candida albicans. The susceptibility tests were done by agar well diffusion method. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of active extracts and sub-fractions were measured using method of National Committee for Clinical Laboratory Standards (NCCLS). RESULTS: Only P. olivieri sub-fractions were found to have fungicidal activity among the other investigated plants. The MIC and MFC was found to be high in petroleum ether, chloroform and ethyl acetate fractions (100 and 200 mg/mL) against the studied pathogenic fungi and the yeast Candida albicans. P. olivieri sub-fractions significantly inhibited the growth of all pathogenic fungi and the yeast studied. CONCLUSION: If the antifungal activity of P. olivieri is confirmed by in vivo studies and if the responsible compound (s) is isolated and identified, it could be a good remedy for mycotic infections.

Biological Effects and Clinical Applications of Dwarf Elder ( Sambucus ebulus L): A Review.

Dwarf elder ( Sambucus ebulus L) is one of the best known medicinal herbs since ancient times. In view of its benefits as a widely applicable phytomedicine, it is still used in folk medicine of different parts of the world. In addition to its nutritional values, dwarf elder contains different phytochemicals among which flavonoids and lectins are responsible for most of its therapeutic effects. Dwarf elder has been used for different ailments including: joint pains, cold, wounds, and infections. Nevertheless, recent evidence has revealed its potentials for making attempts at treating cancer and metabolic disorders. This review aimed to provide a comprehensive description of dwarf elder regarding its traditional uses and modern findings which may contribute to the development of novel natural-based therapeutic agents.

Sambulin A and B, non-glycosidic iridoids from Sambucus ebulus, exert significant in vitro anti-inflammatory activity in LPS-induced RAW 264.7 macrophages via inhibition of MAPKs's phosphorylation.

ETHNOPHARMACOLOGICAL RELEVANCE: The leaves of Sambucus ebulus L. (Adoxaceae) are widely used in Turkish folk medicine particularly against inflammatory disorders. The fresh leaves after wilted over fire or the poultices prepared are directly applied externally to heal burns, edema, eczema, urticarial and abscess. Two iridoids were recently isolated (sambulin A, sambulin B) from the leaves of S. ebulus. AIM OF THE STUDY: This study aims to investigate the in vitro anti-inflammatory activities of these iridoids on LPS-induced RAW 264.7 macrophages. MATERIALS AND METHODS: Raw 264.7 macrophages were treated with 12.5, 25 and 50µg/ml Sambulin A and 6.25, 12.5 and 25µg/ml Sambulin B and induced with 1µg/ml lipopolysaccaharides (LPS). Effect of the compounds on nitric oxide (NO) production and cytokines (TNFα, IL-6) were determined by Griess and ELISA assays respectively. iNOS and the phosphorylation levels of MAPKs (ERK, JNK) were examined by Western Blot. RESULTS: Sambulin A and sambulin B inhibited 52.82% and 72.88% of NO production at 50 and 25µg/ml concentrations respectively. The levels of iNOS were significantly decreased by both molecules, sambulin B at 25µg/ml almost completely decreased iNOS levels (97.53%). Both molecules significantly inhibited TNFα productions. However, only sambulin B inhibited IL-6 production. Consequently, it was shown that sambulin B exerted its effect through the inhibition of ERK and JNK phosphorylations. CONCLUSION: The prominent bioactivities exerted by two iridoids will contribute to explanation of the usage of S. ebulus in traditional medicine against rheumatoid diseases.

Effects of temperature, pH and sugar binding on the structures of lectins ebulin f and SELfd.

Ebulin f and SELfd are two lectins of Sambucus ebulus L. that show different stability and digestibility properties in gastric fluid due to their structural differences which may explain their different toxicological profiles. The main aim was to determine the effects of pH, temperature and sugar binding on the intrinsic structures of both proteins by fluorescence analyses. Quenching experiments were conducted, under different pH and temperature conditions, with acrylamide (uncharged) and iodide (charged), to study the possible changes of their intrinsic fluorescence. Results revealed that the native structure of SELfd is more folded than that of ebulin f. At pH 2.0, ebulin f displayed a more open structure than at neutral pH. It can be concluded that this is the main reason why ebulin f is accessible to pepsin action and more sensitive to degradation, in contrast to SELfd as we reported previously.