Mojtaba Asadollahi

Cytotoxic, antioxidant and antimicrobial effects of nine species of woundwort (Stachys) plants

Abstract Context: Woundwort (Stachys) plants from the Lamiaceae family have been used in folk medicine for various purposes. Objective: This study was designed to analyze cytotoxic, antioxidant and antimicrobial activities of Stachys plants, because these fields have extensively benefited of drug discovery from natural sources. Materials and methods: Nine Stachys plants were collected from different regions of Iran. Cytotoxic activities of methanol, 80% methanol and dichloromethane (DCM) extracts of these plants were assessed on three human cancer cell lines (HL-60, K562 and MCF-7 cells) with the MTT assay, while antioxidant and antimicrobial activities were determined on methanol extracts by DPPH and nutrient broth micro-dilution assays, respectively. Results: DCM extract of St. pilifera Benth. had the lowest IC50 in three cancer cell lines ranging from 33.1 to 48.2 µg/ml, followed by the 80% methanol extract of St. persica S.G.Gmel. ex C.A.Mey. (IC50 range: 62.1–104.1 µg/ml) and DCM extract of St. byzantina C. Koch (IC50 range: 62.7–131.0 µg/ml). St. byzantina. St. lavandulifolia Vahl., St. acerosa Boiss., St. obtusicrena Boiss. and St. persica showed lowest IC50 values in the DPPH scavenging assay (135.1, 162.6, 164.7, 169.4 and 172.4 µg/ml, respectively), while their total phenolic contents were 23.9, 18.2, 18.6, 20.4, 27.8 mg equivalent of gallic acid in 1 g dry plant, respectively. The methanol extracts of St. byzantina and St. persica inhibited all six tested Gram-negative and Gram-positive bacterial strains. Conclusion: Various Stachys species (especially St. byzantina and St. persica) are valuable sources of natural compounds with important biological properties.

Chemical Classification of the Essential Oils of the Iranian Salvia Species in Comparison with Their Botanical Taxonomy

The essential oils of eight Salvia species collected from different localities in Iran were analyzed by gas chromatography/mass spectrometry (GC/MS). The analytical results were compared with those previously published for related Iranian sage species in order to identify chemical markers for these species. Salvia eremophila, S. hypoleuca, and S. reuteriana are endemic, while S. atropatana, S. chloroleuca, S. santolinifolia, S. aegyptiaca, and S. macrosiphon also grow wild in neighboring countries. We categorized the Iranian Salvia species into four main chemotypes according to their essential‐oil constituents: those which are dominated by 1) monoterpenes, 2) mono‐ and sesquiterpenes, or 3) sesquiterpenes as the major constituents, and 4) those containing low‐molecular‐weight acids, aldehydes, and esters, and green‐leaf volatiles (GLVs). Likely due to the chemical diversity of different Salvia chemotypes, this categorization was supported by principal component analysis (PCA) for the group sampled here, but not for the values reported in the literature. We identified the following chemical markers: α‐pinene, β‐pinene, 1,8‐cineol, linalool, and borneol in monoterpene‐rich species, or β‐caryophyllene, germacrene D, bicyclogermacrene, spathulenol, and caryophyllene oxide in sesquiterpene‐rich species. Among these, α‐pinene, β‐caryophyllene, and germacrene D are the most common and abundant in the Salvia species investigated. In accordance with their close biological taxonomy, the chemical similarity of the essential oils of S. santolinifolia and S. eremophila is so high that we may consider them chemically identical.

Carthamus, Salvia and Stachys species protect neuronal cells against oxidative stress-induced apoptosis.

Abstract Context: Finding effective therapies for neurodegenerative diseases is of utmost importance for the aging population. Plants growing in Iran are rich sources of antioxidants and active phytochemicals. Objective: The protective capacity of plants, with a special focus on those with reported antioxidant or neuroprotective potential or nervous system-related applications in folk medicine, was tested against oxidative stress-induced apoptosis. Materials and methods: Aerial parts of 20 plants including Carthamus, Salvia, and Stachys species were extracted with 80% methanol and dichloromethane and preincubated with neuronal PC12 cells for 3 h. Oxidative stress and apoptosis were induced by hydrogen peroxide (75 µM, 1 h exposure). Cell viability and intracellular reactive oxygen species (ROS) were measured by MTT and 2′,7′-dichlorofluorescein-diacetate (DCFH-DA) assays, respectively, while apoptosis was determined by annexin V-FITC/propidium iodide staining by a flow cytometer. Results: Eighty percent methanol extracts of Carthamus oxyacantha Bieb. (Asteraceae), Salvia santolinifolia Boiss. (Lamiaceae), and Salvia sclarea L. (Lamiaceae) at the concentration of 100 μg/ml showed significant neuroprotection in the MTT assay by 38.7, 34.7, and 39.5%, respectively, and inhibited intracellular ROS by 48.6, 61.9, and 61.4%, respectively. The first two extracts also significantly inhibited apoptosis. Dichloromethane extracts of C. oxyacantha and Stachys pilifera Benth. (Lamiaceae) at the concentration of 25 μg/ml showed neuroprotection by 27.5 and 26.5%, respectively, and inhibited ROS by 44.5 and 39.4%, respectively. Conclusion: The above-mentioned plants seem to have important biological activities and their further study may lead to the discovery of new natural therapeutics useful against disorders such as Alzheimer and Parkinson diseases.

Essential oil composition and cytotoxic activity of Ducrosia anethifolia and Ducrosia flabellifolia from Iran

Hydrodistilled volatile oils from the aerial parts of Ducrosia anethifolia and Ducrosia flabellifolia were investigated by gas chromatography (GC) and GC/mass spectrometry (GC/MS). The plants were collected from west and southwest Iran. Thirty-two compounds in D. anethifolia and thirty-three compounds in D. flabellifolia were identified, representing 93.4% and 88.2% of the total oils, respectively. The major components of D. anethifolia oil were dodecanal (28.8%), decanal (21.1%), (2E)-tridecen-1-al (15.8%) and (2E)-dodecenal (13.4%). The most abundant components in essential oil of D. flabellifolia were decanal (32.8%), dodecanal (32.6%), decanol (4.3%) and (2E)-tridecen-1-al (3.3%). Cytotoxic activity assessed on three human cancer cell lines (K562, LS180 and MCF-7) showed that essential oil from D. anethifolia represented good to moderate cytotoxic activity, while D. flabellifolia had a moderate to weak activity.

Essential Oil Composition and Cytotoxic Activity of Libanotis transcaucasica Schischk. from Iran

Background: Libanotis transcaucasica Schischk. belonging to the family of Apiaceae is a perennial plant that is distributed in Europe, northwest of Iran and also Caucasia. Purpose: Due to the interest on development of drugs from natural origins, we studied the components and cytotoxic activity of essential oil from L. transcaucasica. Methods: The essential oil of L. transcaucasica was obtained by hydro distillation of its air-dried aerial parts and was analyzed by gas chromatography (GC-FID) and gas chromatography-mass spectrometry (GC-MS). Cytotoxic activity of the essential oil was assessed on four human cancer cell lines (HeLa, LS180, MCF-7 and Raji). Results: The yield of oil was 0.9% (w/w) and it had yellow color. Fifty-four compounds in L. transcaucasica representing 84.0% of the oil components were identified. Germacrene B (20.2%) was the most abundant compound in this oil, followed by isospathulenol (11.0%), germacrene D (9.2%) and kessane (5.5%). The volatile oil displayed weak to moderate cytotoxic activity in all evaluated human cancer cell lines (IC50 =0.9-0.2 mg/ml). The highest and the lowest cytotoxic effects were observed on Raji and LS180 cell lines, respectively. Conclusion: Sesquiterpene hydrocarbons were identified as the main components of the essential oil (48.3%). The cytotoxic activity observed in the essential oil may be contributed to the existence of this group of hydrocarbons in the plant.

Ecological Roles and Biological Activities of Specialized Metabolites from the Genus Nicotiana

Species of Nicotiana grow naturally in different parts of the world and have long been used both medicinally and recreationally by human societies. More recently in our history, Nicotiana tabacum has attracted interest as one of the most economically important industrial crops. Nicotiana species are frequently investigated for their bioactive natural products, and the ecological role of their specialized metabolites in responses to abiotic stress or biotic stress factors like pathogens and herbivores. The interest of tobacco companies in genetic information as well as the success of a few wild tobacco species as experimental model organisms have resulted in growing knowledge about the molecular biology and ecology of these plants and functional studies of the plants natural products. Although a large number of reviews and books on biologically active natural products already exists, mostly from N. tabacum, we focus our attention on the ecological roles and biological activity of natural products, versus products from cured and processed material, in this Review. The studied compounds include alkaloids, aromatic compounds, flavonoids, volatiles, sesquiterpenoids, diterpenes alcohols, and sugar esters from trichomes of the plants, and recently characterized acyclic hydroxygeranyllinalool diterpene glycosides (HGL-DTGs). In this Review (1800s-2017), we describe the above-mentioned classes of natural products, emphasizing their biological activities and functions as they have been determined either in bioassay-guided purification approaches or in bioassays with plants in which the expression of specific biosynthetic genes has been genetically manipulated. Additionally, a review on the history, taxonomy, ecology, and medicinal application of different Nicotiana species growing around the globe presented in this Review may be of interest for pharmacognosists, natural products, and ecological chemists.

Antimicrobial Survey of Local Herbal Drugs against Acinetobacter baumannii Isolated from Patients Admitted to a Level-I Trauma Center

Objective: To determine the antimicrobial activity and entity of several local herbal plants against Acintobacters isolated from trauma patients admitted to a Level-I trauma center. Methods: The antibacterial activities of the Satureja bachtiarica oil and someselected Iranian medicinal plants (Artemisia sieberi and Tanacetum dumosum belonging to the Asteraceae/Compositae; Salvia mirzayanii and Mentha mozaffarianii belonging to the Lamiaceae/Labiatae) were assayed on A. baumannii by microdilution and agar disc diffusion methods. Having obtained the acceptable antibacterial data, the shade-dried aerial parts of the plants were extracted by hydrodistillation method using Clevenger apparatus according to European pharmacopeia for 3 h. The analysis of S. bachtiarica essential oil accompanied by other herbal drug oils were performed by using GC/FID and GC/MS methods. Results: Outcomes revealed that the S. bachtiarica essential oil exhibited the potent antibacterial capability against Acinetobacter strains in comparison with Colistin, as a positive control. For S. bachtiarica,the growth inhibition zone and minimum inhibitory concentration (MIC) values were 21 mm and 0.5 mg/ml, while, for Colistin, the data were in order: 8 mm and 0.016 mg/ml. Consequently, GC/MS outcomes demonstrated that the major components of the essence were carvacrol (48.6%), followed by p-Cymene (16.6%), γ-terpinene (6.9%) and linalool (5.3%). Conclusion: Based on the considerable inhibitory activity against nosocomial infections by essential oil of S. bachtiarica, it could be considered as the suitable candidate in the food industry and pharmaceutical uses.

Comparison Between Head-Space SPME and Hydrodistillation-GC-MS of the Volatiles ofThymus daenensis

Abstract The volatile constituents of Thymus daenensis growing wild in surrounding Shiraz- Iran were extracted by hydrodistillation (HD) and Head-Space Solid-Phase Micro Extraction (HS-SPME) methods and further analyzed by gas chromatography mass spectroscopy (GC-MS). In the HS-SPME and essential oil (EO) of T. daenensis, 40 and 28 components were identified respectively, which represented by 98.0 % and 99.7 % of the total detected components. The major volatile components in the HS and EO were thymol (9.1±0.83, 52.2 %), carvacrol (20.25±2.38, 37.4 %) and β-caryophyllene (17.98±2.19, 5.6 %) respectively. The concentration of thymol in the HS of the plant was determined to be 224±46 ng/L. The present paper shows similar major constituents detected by HS-SPME and HD-GC-MS, but the levels of the compounds are different in the above-mentioned extraction methods.

ESSENTIAL OIL OF Tanacetum dumosum AS A NEW SOURCE OF FRAGRANOL

Tanacetum (tansy) belongs to the tribe Anthemideae, which is one of the largest tribes of Asteraceae family, comprising about 111 genera and ca. 1800 species worldwide [1]. In the flora of Iran, this genus is represented by 31 annual and perennial species, of which 15 are endemic [2]. The essential oils of several Iranian tansies, including T. bachtiaricum Mozaff. [3], T. fruticulosum Ledeb. [4], T. canescens and T. turcomanicum [5], and T. polycephalum Schultz-Bip. [6], were analyzed by different authors and found to have monoterpenoids as major constituents. Among them, pinenes, 1,8-cineol, camphor, chrysanthenol derivatives, and thujones are detected more frequently. In addition to the above regular monoterpenoids, irregular monoterpenes such as artemisia ketone, santolina triene, and yomogi alcohol are found in some tansy species, including T. kotschyi [7] and T. santolinoides [8]. To the best of our knowledge, this is the first report on characterization of the chemical composition of the essential oil of T. dumosum. The aerial parts of Tanacetum dumosum Boiss. were collected from Dena mountain at an altitude of 3200 m in July 2011. The plant material was identified by one of us (MA), and a voucher specimen (No. PC-87-113) was deposited at the herbarium of our institute (MNCRS), Shiraz, Iran. The essential oil of the leaves and flowers of the plant (70 g) was extracted by hydrodistillation to yield 0.2 mL of pale-yellow oil, dried over anhydrous sodium sulfate, and analyzed by gas chromatography-mass spectrometry (GC-MS) and GC-flame ionization detector (GC-FID). Twenty seven monoand sesquiterpenoids were identified in the oil using GC retention indices (RRI) and mass spectral (MS) data, obtained from GC and GC-MS experiments, and comparison of these data to those reported for the standard volatiles in the literature [9, 10] (Table 1). Among them, the monoterpene hydrocarbons santolina triene (2.2%) and -pinene (2.2%) and the oxygenated monoterpenes fragranol (26.7%), 1,8-cineol (5.7%), fragranyl acetate (23.6%), and trans-carvyl acetate (10.1%) were detected as the major constituents. Helifolenol A, -sesquiphellandrene, caryophyllene oxide, and germacrene D are the major sesquiterpenes detected in this oil, but they are still in smaller quantities compared to the monoterpenoids fraction (Table 1). Fragranol, the E isomer of 2-isopropenyl-1-methylcyclobutane ethanol, was reported previously in the oils of Achillea santolina [9], A. nobilis [11], and A. falcata [12], but to the best of our knowledge for the first time in tansy species. Previously, fragranol was reported by one of us (ARJ) in the oil of Artemisia haussknechtii [13], a member of Anthemideae, which grows nearby T. dumosum. Grandisol, the Z-isomer of fragranol, is reported to be an aggregation and sex-attractant pheromone for several weevil insects [14, 15], and recently together with camphor and 1,8-cineol as the major constituent of an anti-insect essential oil of Artemisia vestita [16]. The antibacterial and insecticidal activity of the essential oils of the above plant species with similar chemical composition reported here for T. dumosum suggests that this plant is a potential medicinal plant to be explored in the future [9, 11, 12]. The good smell of the plant may be due to the presence of fragranol and its acetate ester and suggests that this plant can be used in the perfume industry. GC-MS and FID Analyses. GC-MS analysis was carried out on an Agilent 7890A GC coupled to an HP-6890 mass spectrometer operating in EI mode at 70 eV. The GC was equipped with a J&W DB-5 ms Scientific column (30 m 0.25 mm i.d., 0.25 m film thickness). The oven temperature was programmed from 60 C to 220 C at 5 C/min and kept for 10 min at 220 C. The carrier gas was helium (He) with a flow rate of 1 mL/min, and the injector temperature was set at 260 C in the split mode for the pure essential oil (1:100). The injection volume was 0.1 L. The GC-FID analysis was performed using the above instrument coupled to a FID with the same analytical conditions, but the column was replaced by a DB1 column with the same dimension as that used in GC-MS with the temperature of the FID set at 260 C.