Inula Oculus-christi Research Articles

Eco-benevolent synthesis of ZnO-NPs and ZnO-MFs from Inula oculus-christi L. (Asteraceae) with effective antioxidant, antimicrobial, DNA cleavage, and decolorization efficiencies.

As a result of the changes occurring globally in recent years, millions of people are facing challenging and even life-threatening diseases such as cancer and the COVID-19 pandemic, among others. This phenomenon has spurred researchers towards developing and implementing innovative and environmentally friendly scientific methods, merging disciplines with significant technological potential, such as nanotechnology with medicinal plants. Therefore, the focus of this research is to synthesize zinc nanoparticles (ZnO-NPs) and microflowers (ZnO-MFs) using extracts of the medicinal plant I. oculus christi prepared in n-hexane and methanol as new bioreduction and capping agents through a simple and environmentally friendly chemical approach. Optical, thermal, and morphological structural analyses of ZnO-NPs and ZnO-MFs were conducted using Ultraviolet-Visible (UV-Vis) spectroscopy, Fourier Transform Infrared (FT-IR) spectroscopy, Thermogravimetric Analysis (TGA), and Field Emission Scanning Electron Microscopy (FE-SEM). Metabolic profiles of extracts from different plant parts were analyzed using Gas Chromatography-Mass Spectrometry (GC-MS) and supported by visualization of contents through Principal Component Analysis (PCA), hierarchical cluster analysis heatmaps, and Pearson correlation graphs. Interestingly, ZnO-NPs and ZnO-MFs exhibited strong antioxidant properties and demonstrated particularly potent antimicrobial activity against Micrococcus luteus NRRL B-4375, Escherichia coli ATCC 25922, and Candida albicans ATCC 10231 strains compared to standard antibiotics. Furthermore, ZnO-NPs and ZnO-MFs showed excellent plasmid DNA-cleavage activity of pBR322 with increasing doses. The photocatalytic performance of the synthesized ZnO-NPs and ZnO-MFs was evaluated for methylene blue (MB), congo red (CR), and safranin-O(SO) dyes, demonstrating remarkable color removal efficiency. Overall, the results provide a promising avenue for the green synthesis of ZnO-NPs and ZnO-MFs using I. oculus-christi L. inflorescence and pappus extracts, potentially revolutionizing biopharmaceutical and catalytic applications in these fields.

Gaillardin inhibits autophagy and induces apoptosis in MCF-7 breast cancer cells by regulating JAK/STAT pathway.

Gaillardin is a potent anti-cancer sesquiterpene lactone found in Inula oculus-christi. The present study examined the effects of gaillardin on apoptosis and autophagy in the MCF-7 breast cancer cell line. The MTT assay was used to unravel the antiproliferative effects of gaillardin on MCF-7 cells. The expression of apoptosis-related genes including CASP3, BAX, BCL2, STAT3, and JAK2, and key markers of autophagy such as ATG1, ATG4, ATG5, ATG7, ATG12, BECN1, and MAP1LC3A were measured by real time-PCR method. The protein expression of Caspase 3, phosphorylated JAK2, phosphorylated STAT3, ATG1, ATG4, ATG5, ATG12, Beclin1, and LC-III was determined using western blotting. Gaillardin treatment significantly decreased the proliferation of MCF-7 cells with a parallel upregulation of the level of pro-apoptotic caspase-3 enzyme with no effect on Bax and Bcl2 expression. The levels of phosphorylated and active forms of JAK2 and STAT3 proteins were reduced following the treatment of MCF-7 cells with gaillardin. This sesquiterpene lactone com-pound considerably downregulated the levels of six autophagy markers, including ATG1, ATG4, ATG5, ATG12, Beclin1, and LC-III in MCF-7 cells. These data indicated the apoptosis-inducing activity of gaillardin in MCF-7 cells by a mechanism that inhibits the JAK/STAT signaling pathway. Further, autophagy inhibition was the other phenomenon caused by gaillardin in MCF-7 cells. These results can provide evidence to highlight the role of gaillardin as a novel therapeutic for the treatment of breast cancer.

Phenolic profile, antioxidant and anticandidal activities of Inula oculus-christi L. from Turkey

Phenolic profile, antioxidant and anticandidal activities of Inula oculus-christi L. from Turkey

Gaillardin, a potent sesquiterpene lactone induces apoptosis via down-regulation of NF‐κβ in gastric cancer cells, AGS and MKN45

Ethnopharmacological relevanceThe genus Inula has been traditionally used as folk medicine in treating different illnesses such as kidney stones, urethra infection, jaundice, bronchitis, respiratory diseases and cancer. Aim of the studyGaillardin, a sesquiterpene lactone isolated from Inula oculus-christi, seems to have great potential as an anti-cancer agent. This study was carried out to evaluate the anti-cancer properties of Gaillardin in gastric cancer cells and also its possible underlying mechanism. MethodsThe colorimetric MTT assay was used to assess metabolic activity of cells as an indicator of viability and cytotoxicity. Flow cytometry using Annexin V-FITC/PI was applied to detect and quantify the level of apoptosis. Detection of activated caspase-3, as a biochemical marker of apoptosis, was done using caspase-3 assay kit. Activation of NF-κB pathway was determined by western blotting. The mRNA expression levels of NF-κB target genes were measured using quantitative RT-PCR. Moreover, intracellular reactive oxygen species (ROS) production was evaluated. ResultsGaillardin significantly reduced cell viability in a time and dose-dependent manner. The inhibitory effect of Gaillardin was attributed to induction of apoptosis. Investigation about the underlying mechanism revealed that Gaillardin exerts its action through inhibition of NF-κB activation and subsequently down-regulation of genes (COX-2, MMP-9, TWIST-1, and BCl-2) regulated by NF-κB. Moreover, Gaillardin caused remarkable elevation in ROS production in AGS and MKN45 cells. ConclusionWe provided evidences about the role of NF-κB pathway in the induction of apoptosis by the sesquiterpene lactone Gaillardin in AGS and MKN45 cells for the first time, which suggest Gaillardin could be considered as a promising natural agent for further investigations to identify new potent anticancer drugs in the future.

STUDY OF AMINO ACID COMPOSITION IN ABOVE-GROUND PART INULA OCULUS CHRISTI L.

One of the conditions for the introduction of a new type of medicinal plant raw materials into medical practice is a comprehensive study of its chemical composition. The wide distribution of amino acids in plants and their high biological activity contribute to the effective action of medicinal raw materials and preparations derived from it on the body. Amino acids give other biologically active substances an easily digestible form, while simultaneously potentiating their pharmacological effect. The world around us contains many amino acids, but about 20 are enough for people to function normally. Amino acids as constituents of proteins are involved in all life processes along with nucleic acids, carbohydrates and lipids. In addition to amino acids that make up proteins, living organisms have a constant reserve of free amino acids contained in tissues and cell sap. They are in dynamic equilibrium with numerous exchange reactions. Amino acids are used in the biosynthesis of phosphatides, porphins, nucleotides. The qualitative and quantitative composition of amino acids in the aerial part of Inula oculus christi was studied. The analysis was carried out on an AAA400 amino acid analyzer, a highly specialized automated liquid chromatograph with computer control, equipped with a post-column detector system. 0.2 g of a sample (accurately weighed on an analytical balance) was suspended in a 20 ml penicillin vial, added to the top with normal hydrochloric acid, tightly closed with a rubber lid, and placed in an oven for 23 hours at 110°C. After hydrolysis, it was cooled to room temperature and evaporated to dryness in a rotary evaporator, after which 5 ml of purified water was added and evaporated again. This water wash procedure is necessary to get rid of residual hydrochloric acid, which negatively affects yield and peak separation. To the sample evaporated to dryness, 50 ml of loading buffer (pH 2.2) was added. Before injection into the ion exchange column, the sample was filtered through a 0.2 µm syringe filter or blue ribbon filter paper. As a result of the analysis, 16 amino acids were found, of which 7 a.re essenti.al (valine, isoleucine, leucine, lysine, methionine, threonine, phenylalanine) and 9 nonessential (alanine, arginine, aspartic acid, histidine, glycine, glutamic acid, proline, serine, tyrosine).

Caffeoylquinic Acids, Cytotoxic, Antioxidant, Acetylcholinesterase and Tyrosinase Enzyme Inhibitory Activities of Six Inula Species from Bulgaria.

Chlorogenic (5-CQA), 1,5-, 3,5-, 4,5- and 3,4-dicaffeoylquinic (DCQA) acids were identified and quantified in the methanol extracts of Inula oculus-christi L., I. bifrons L., I. aschersoniana Janka var. aschersoniana, I. ensifolia L., I. conyza (Griess.) DC. and I. germanica L. by HPLC analysis. The amount of 5-CQA varied from 5.48 to 28.44 mg/g DE and the highest content was detected in I. ensifolia. 1,5-DCQA (4.05-55.25 mg/g DE) was the most abundant dicaffeoyl ester of quinic acid followed by 3,5-DCQA, 4,5-DCQA and 3,4-DCQA. The extract of I. ensifolia showed the highest total phenolic content (119.92±0.95 mg GAE/g DE) and exhibited the strongest DPPH radical scavenging activity (69.41±0.55 %). I. bifrons extract was found to be the most active sample against ABTS.+ (TEAC 0.257±0.012 mg/mL) and the best tyrosinase inhibitor. The studied extracts demonstrated a low inhibitory effect towards acetylcholinesterase and possessed low cytotoxicity in concentration range from 10 to 300 μg/mL toward non-cancer (MDCK II) and cancer (A 549) cells.

Ergolide, a potent sesquiterpene lactone induces cell cycle arrest along with ROS-dependent apoptosis and potentiates vincristine cytotoxicity in ALL cell lines

Ethnopharmacological relevanceInula oculus christi belongs to the family of Asteraceae and it was traditionally wide used in treatment of kidney stones and urethra infection; besides, recently the potent sesquiterpene lactones isolated from inula species has gained increasing attention in cancer treatments. This study investigates the anti-cancer properties and underlying mechanism of ergolide isolated from Inula oculus christi against leukemic cell lines. MethodsViability, metabolic activity and proliferation evaluated using different index of MTT assay such as IC50 and GI50. Human erythrocytes were used to evaluate hemolytic activity. Flow-cytometry was used to detect and measure ROS level, and the induction of apoptosis and autophagy were evaluated using Annexin V/PI, Acridine Orange staining, respectively. Moreover, qRT-PCR was performed to examine the expression of a large cohort of crucial regulatory genes. Tunel assay was also carried out to assess morphologically ergolide effects. ResultsErgolide did not exert ant cytotoxicity against non-tumorous cells and did not cause noticeable hemolysis. It also caused ROS production during early hours after treatment of cells which was then followed by cell cycle arrest in G0/G1 phase and autophagy induction. Using N-acetyl-L-cysteine (NAC), we found that ergolide could not increase ROS and induce autophagy and moreover repressed cell death, indicating that ergolide induce cell death through ROS-dependent manner by altering the expression of pro apoptotic related genes. Autophagy inhibition also potentiated ergolide-induced cell death. Furthermore, ergolide intensified vincristine cytotoxicity against acute lymphoblastic leukemia (ALL) cell lines revealed robust synergistic properties of ergolide with VCR. ConclusionHere we showed that ergolide could be considered as a potent natural compound against leukemic cells by inducing cell cycle arrest followed by dose-dependent cell death. Based on results, Autophagy response in a result of ROS accumulation acted as a survival pathway and blocking this pathway could noticeably increase ergolide cytotoxicity on ALL cell lines.

Evaluation of Anticonvulsant Effect of Aqueous and Methanolic Extracts of Seven Inula Species.

In Iranian traditional medicine, Inula species have been used for the treatment of seizure. In this study we decided to investigate the anticonvulsant activity of seven species from this genus to find an effective remedy for seizure with less adverse effects compared to the available medicines. Aqueous and methanolic extracts of Inula britannica, Inula helenium, Inula viscidula, Inula oculus-christi, Inula aucheriana, Inula thapsoides, and Inula salicina were prepared and their antiepileptic activity was investigated by maximal electroshock (MES) and pentylentetrazole (PTZ) tests on Male NMRI Albino mice. Diazepam was used as positive control in both tests. In addition, two extracts with the best anticonvulsant activities were selected and their sedative and hypnotic effects were evaluated using open field and righting reflex tests, respectively. The effects of the both extracts on memory and motor coordination were also assessed by step-through passive avoidance and rotarod tests, respectively. Aqueous extract of Inula britannica and Inula viscidula showed the best activity in MES model and their ED50 (with 95% confidence interval) was 19.5 (7.9~48.5) mg/kg and 12.7(10.0~16.3) mg/kg, respectively. None of the extracts showed noticeable anticonvulsant effects in the PTZ model. The active extracts also showed sedative-hypnotic effects in righting reflex and open field tests. Furthermore, both extracts did not affect the memory and motor coordination in the experimental models. The anticonvulsant and sedative activities of the extracts were antagonized by flumazenil, indicating that benzodiazepine receptors are probably involved in the effects. This study indicates that Inula britannica and Inula viscidula are good candidates for further phytochemical and mechanistic studies in order to find anticonvulsant and sedative-hypnotic compounds with less adverse effect on memory and motor coordination.

New sesquiterpene lactones from Inula oculus-christi L.

Investigation of the aerial parts of Inula oculus-christi L. led to the identification of four new sesquiterpene lactones: an eudesmanolide (4α,15α-epoxypulchellin E) and three guaianolides (2α-acetoxy-4α,9β-dihydroxy-1β-guaia-11(13),10(14)-dien-12,8α-olide, 9β,10β-epoxygaillardin, 9α,10α-epoxy-2-epi-gaillardin), in addition to the known gaillardin, pulchellin E, pulchellin C and 3-O-palmitates of 16β-hydroxylupeol, 16β-hydroxy-β-amyrin, and faradiol. The structures of all compounds have been elucidated on the basis of their spectral data.

Biosynthesis of very long-chain fatty alcohols and wax esters in metabolically engineered strains of Saccharomyces cerevisiae

Investigation of the aerial parts of Inula oculus-christi L. led to the identification of four new sesquiterpene lactones: an eudesmanolide (4α,15α-epoxypulchellin E) and three guaianolides (2α-acetoxy-4α,9β-dihydroxy-1β-guaia-11(13),10(14)-dien-12,8α-olide, 9β,10β-epoxygaillardin, 9α,10α-epoxy-2-epi-gaillardin), in addition to the known gaillardin, pulchellin E, pulchellin C and 3-O-palmitates of 16β-hydroxylupeol, 16β-hydroxy-β-amyrin, and faradiol. The structures of all compounds have been elucidated on the basis of their spectral data.

Anatomical and micro-morphological analysis of the fruit and vegetative organs of Inula oculus-christi L. in the Pannonian part of Serbia

Inula oculus-christi L. is a perennial plant, 20-60 cm high, growing in south-east Europe, central and southern Russia and some parts of Asia. It belongs to Pontic-Pannonian floristic element. In Pannonian part of Serbia this species is extremely rare, with very few recently confirmed literature and herbarium data. According to the field data for the period from 2013 to 2015, only one population of this species was recorded in Pannonian part of Serbia, on the site of Rimski Sanac, which is threatened by anthropogenic activities. Furthermore, anatomical and micro-morphological features of I. oculus-christi have not been documented so far. Therefore, in order to get better knowledge of the biology this species, as well as on the basis of the abovementioned, its anatomical and micro-morphological characteristics were investigated. Cross sections of the leaf, stem, rhizome and fruit were obtained using cryotechnique procedure by Leica CM 1850 cryostat. Using a light microscopy, detailed descriptions of anatomical characteristics of the analysed organs were given. The scanning electron microscopy revealed specific qualitative features of leaf and fruit that characterize the species. Obtained data may be useful in determination of this species and represent the valuable contribution to its micro-morphological and anatomical differentiation from other congeneric and related species.

Natural sesquiterpen lactones as acetylcholinesterase inhibitors.

Background and the purpose of the study: The amount of elder people who suffer from Alzheimer disease is continuously increasing every year. Cholinesterase inhibitors have shown to be effective in alleviating the symptoms of the disease, thus opening a field of research for these treatments. Herbal products, owning a reputation as effective agents in many biological studies are now drawing attention for inhibiting acetylcholinesterase, in other words, Alzheimer disease. In the present study, the ability of three sesquiterpene lactones from Inula oculus-christi and I. aucheriana to inhibit AChE has been evaluated through Ellman assay. Materials and Methods: Gaillardin and pulchellin C were obtained from I. oculus-christi and britannin from I. aucheriana by chromatographic methods. They were dissolved in methanol in concentration of 3 mg/mL and the AChEI activity of the compounds was determined by Ellman method using Acethylthiocholine iodide as the substrate and 5, 5'-dithiobis-2-nitrobenzoic acid as the reagent, in 96-well plates at 405 nm. Results: AChEI activity of the examined compounds was obtained as 67.0, 25.2 and 10.9% in concentration of 300 µg/L for gaillardin, britannin and pulchellin C, respectively. Conclusion: Among the three sesquiterpene lactones, gaillardin with 67% inhibition of AChE could be considered a good candidate for future Alzheimer studies.

Cytotoxic Activity and Apoptosis Induction by Gaillardin

Cytotoxic activity of gaillardin, a sesquiterpene lactone isolated from Inula oculus-christi L. (Asteraceae), was assessed in the human breast adenocarcinoma cell line MCF-7, human hepatocellular carcinoma cell line HepG-2, human non-small cell lung carcinoma cell line A-549, and human colon adenocarcinoma cell line HT-29, resulting in IC50 values of 6.37, 6.20, 4.76, and 1.81 microg/mL, respectively, in the microculture tetrazolium-formazan MTT assay. In vitro apoptosis-inducing properties of gaillardin were also evaluated in MCF-7 cells with the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) assay. The results suggest gaillardin as a candidate for further studies in cancer therapy

A new species of Tephritis Latreille (Diptera: Tephritidae) from Turkey

Tephritis ozaslani sp. nov. is described from Artvin Province of Turkey and placed in the T. pulchra species group. The key to the T. pulchra group is provided, which includes 9 species. The main characters that distinguish the new species from the closely resembling Palaearctic species, T. cometa israilis Freidberg, T. acanthiophilopsis Hering, T. erdemlii Kütük, T. divisa Rondani, T. recurrens Loew, T. merzi Freidberg & Kütük, and T. hurvitzi Freidberg, are discussed. In addition, Inula oculus christi L. (Asteraceae) is determined as a host plant of the new species.

Screening of the in vitro amoebicidal activities of Pastinaca armenea (Fisch. & C.A.Mey.) and Inula oculus-christi (L.) on Acanthamoeba castellanii cysts and trophozoites

The aim of this study is to evaluate the in vitro amoebicidal activity of the aqueous extracts of Pastinaca armenea and Inula oculus-christi from Turkey. In the presence of aqueous extracts (ranging from 1.0 to 32.0 mg/ml), numbers of the viable Acanthamoeba castellanii trophozoites and cysts were decreased during the experimental process. Both extracts showed a time- and dose-dependent amoebicidal action on the trophozoites and cysts. Among the extracts tested, I. oculus showed the strongest amoebicidal effect on the trophozoites and cysts. In the presence of 32.00 mg/ml extract solution in the media, no viable trophozoites were determined from the time of 24 h. Moreover, in the presence of 16.00 mg/ml I. oculus-christi extract, no viable trophozoites were detected in 72 h of the experiment. Effectiveness of I. oculus-christi extract was found moderate against the cysts. In the presence of 32.00 mg/ml extract, only 25.3% of the total cysts were killed effectively. In the case of P. armenea, 40.3% and 23.0% of the trophozoites and cysts were killed in the presence of 32.00 mg/ml extract at the end of the experimental process (72 h), respectively.

Constituents of the Volatile Oil of Inula oculus-christi L. from Iran

The essential oil of Inula oculus-christi was examined by GC and GC/MS. Sixty compounds were identifed representing 90.6% of the oil. The major components were pentacosane (13.7%), palmitic acid (13.6%), dill apiole (11.4%), methyl eugenol (9.6%) and viridiforol (5.7%).

Sesquiterpene lactones from the aerial parts of Inula oculus-christi

Pulchellin E ( 1) and gaillardin ( 2) were isolated from the aerial parts of Inula oculus-christi, along with the flavone hispidulin. The 13C-NMR chemical shifts of 1 and 2 are reported.