Genetic variation and structure of endemic and endangered wild celery (Kelussia odoratissima Mozaff.) quantified using novel microsatellite markers developed by next-generation sequencing.

Kelussia odoratissima Mozaff. (Apiaceae) is a native plant that has been traditionally consumed in Iran's food and pharmaceutical industries. Overharvesting of the taxon, especially at the beginning of the growing season, due to its considerable medicinal and economic value, is believed to be the main reason for the extirpating of this plant. The consequences of the severe anthropogenic impacts on the genetic diversity of populations are poorly known. In order to investigate the level of genetic variation and patterns of the genetic structure of K. odoratissima, we developed novel microsatellite markers using the 454 Roche next-generation sequencing (NGS) platform for the first time. Out of 1,165 microsatellite markers bioinformatically confirmed, twenty-five were tested, of which 23 were used to screen genetic variation across 12 natural populations. Our results showed that the average number of alleles per locus and the polymorphic information content (PIC) were 10.87 (range 7 to 27), and 0.81 (range 0.67 to 0.94), respectively. The mean observed and expected heterozygosities (± SD) across all populations were 0.80 ± 0.31 and 0.72 ± 0.14, respectively. The average pairwise FST among the populations was 0.37 (range 0.04 to 0.81). Bayesian and distance-based clustering, and principal coordinate analyses revealed at least four major genetic clusters. Although high level of structure can be explained by landscape topography and geographic distance, presence of admixed populations can be associated to seed or pollen dispersal. Contrary to expectations, the high level of genetic variation and lack of inbreeding suggest that overexploitation has not yet significantly purged the allelic variability within the natural populations in protected areas.

The essential oil from Kelussia odoratissima Mozaff. fruits exerting potent analgesic and anti-inflammatory effects / El aceite esencial de frutos de Kelussia odoratissima Mozaff. ejerce potentes efectos analgésicos y antiinflamatorios

Kelussia odoratissima fruits are utilized in Persian traditional medicine as a painkiller and to prevent inflammation-based disorders. Considering the higher content of essential oil in the fruits, the oil's anti-inflammatory and analgesic activities were investigated via the paw edema triggered in mice and the writhing test and hot plate technique, respectively. It was observed that the 100, and 200 mg/Kg doses of the oil revealed an analgesic impact (p<0.001) considering the increment in the reaction time needed for the hot plate approach. Furthermore, 100 and 200 mg/Kg doses of the oil caused a reduction in the frequency of writhes in the mice (p<0.01 and p<0.001, respectively). Using all examined doses of theoil (25, 50, and 100 mg/Kg) caused inflammatory reduction (p<0.001). The findings indicated that the oil possess significant activities against acute inflammation. It had both peripheral and central pain-killing impacts. The main components 3-n-butylphthalide (28.3%) and germacrene D (17.3%) can be considered as the responsible compounds to manage the inflammation and pain.

Las frutas de Kelussia odoratissima se utilizan en la medicina tradicional persa como analgésico y para prevenir los trastornos basados en la inflamación. Teniendo en cuenta el mayor contenido de aceite esencial en las frutas, se investigaron las actividades antiinflamatorias y analgésicas del aceite a través del edema de la pata desencadenado en ratones y la prueba de contorsiones y la técnica del plato caliente, respectivamente. Se observó que las dosis de 100 y 200 mg / kg del aceite revelaron un impacto analgésico (p<0,001) considerando el incremento en el tiempo de reacción necesario para el enfoque de placa caliente. Además, dosis de 100 y 200 mg / kg del aceite provocaron una reducción en la frecuencia de retorcimientos en los ratones (p<0,01 y p<0,001, respectivamente). El uso de todas las dosis examinadas del aceite (25, 50 y 100 mg/kg) provocó una reducción inflamatoria (p<0,001). Los hallazgos indicaron que el aceite posee actividades significativas contra la inflamación aguda. Tiene impactos analgésicos tanto periféricos como centrales. Los principales componentes 3-n-butilftalida (28,3%) y germacreno D (17,3%) pueden considerarse como los compuestos responsables del manejo de la inflamación y el dolor.

Determining Effective Environmental Factors in the Distribution of Endangered Endemic Medicinal Plant Species Using the BMLR Model: The Example of Wild Celery (Kelussia odoratissima Mozaff., Apiaceae) in Zagros (Iran).

Kelussia odoratissima Mozaff. is a medicinal species native to Iran. The goal of this research was to determine the environmental factors important for the distribution of K. doratissima in Iran using BMLR modeling. Six random transects were established throughout the species' habitat, and 220 quadrats with an area of 4 m2 were plotted. The canopy cover percentages of K. doratissima were estimated in each quadrat. Topographic factors, including elevation, slope, and aspect maps, were generated by creating DEM images. Land use, land evaluation, evaporation, temperature, and precipitation maps of the area were created accordingly. The data collected from the experiments were analyzed using the Minitab and R statistical packages. To determine the effect of the studied factors in the distribution of K. doratissima, we ran a set of backward multiple linear regressions. The results showed that the effects of evaporation, elevation, and slope were significant in the species' distribution, with elevation having a positive effect and evaporation and slope showing negative effects. Further, elevation had the highest effect on distribution (greatest absolute value of beta at 9.660). The next most significant factors in the plant's distribution were evaporation (beta = 8.282) and slope (beta = 0.807), respectively.

The Evaluation and Comparing of Cytotoxic Effects of Ferula gummosa Gum, Scutellaria lindbergii, Kelussia odoratissima and Artemisia kopetdaghensis Extracts on ACHN Cell Line.

Renal cell carcinoma (RCC) is one of most fatal cancers. In most patients it is resistant to chemotherapy. Ferula gummosa gum, Scutellaria lindbergii, Kelussia odoratissima, and Artemisia kopetdaghensis are herbs about which there are some cytotoxic activity reports. In this study, cytotoxic and apoptotic activity of these four extracts on RCC cell line (ACHN) were evaluated and compared (ACHN) cells were treated with different concentrations of herbal extracts (15-500 µg/mL). Cell proliferation was determined after 24, 48, and 72 h. by MTT assay. Apoptotic cells were determined using PI staining of DNA fragmentation by flow cytometry. Cell viability decreased with all herbal extracts in ACHN cells by 24, 48, and 72 h. as compared with control. Extracts induced a sub-G1 peak in flow cytometry histogram of treated cells indicating apoptotic cell death is involved in extracts induced-toxicity. Results imply that four herbal extracts inhibit the growth of ACHN cells as a concentration- and time-dependent manner. Also, results show that apoptosis is proposed as the possible mechanism of action. So, four herbal extracts could be considered as good anticancer agents in RCC after further studies.

Phytochemical Analysis and Cytotoxicity Evaluation of Kelussia odoratissima Mozaff.

BACKGROUND: Kelussia odoratissima Mozaff. (Apiaceae) is an edible, indigenous, and ethnomedicinal plant that grows only in Iran. Although antioxidant and anti-inflammatory properties of K. odoratissima have been reported, cytotoxic activity of this plant has not been investigated previously. OBJECTIVE: This study aims to evaluate the cytotoxicity of K. odoratissima leaf extract against a panel of human cancer cell lines. A secondary aim was to perform a phytochemical analysis of the plant's leaf oil. METHODS: To extract the plant oil, dried leaves were subjected to hydrodistillation using a Clevenger-type apparatus for up to 3 hours. For the phytochemical analysis, essential oil was subjected to gas chromatography/mass spectrometry. Plant extraction was performed by macerating leaf powder of K. odoratissima (50 g) in 70% methanol (500 mL) at room temperature (25-28°C) for 24 hours. To perform cytotoxicity assays, methanolic extract of K. odoratissima was tested against a panel of cell lines including MDA-MB468 (human breast cancer cell line), K562 (human leukemia cell line), SKOV3 (human ovarian cancer cell line), Y79 (human eye cancer cell line), A549 (lung cancer cell line), and HEK 293 (normal human embryonic kidney cell line). RESULTS: Gas chromatography/mass spectrometry analysis revealed that sesquiterpens are dominant volatile components of the plant, followed by phthalides comprising 3-butyldine phthalide and 3-n-butyl phthalide, the latter compound being the major component of the leaf oil (25.1%). The leaf extract showed selective and dose-dependent cytotoxicity against MDA-MB468, K562, SKOV3, Y79, and A549 cancer cell lines with IC50 values (concentration that inhibits cell growth by 50%) of 85 µg/mL, 70 µg/mL, 120 µg/mL, 82 µg/mL, and145 µg/mL, respectively. CONCLUSIONS: The present results suggest a direct cytotoxic activity of K. odoratissima leaf extract against human cancer cell lines. This activity of K. odoratissima may find application in combination with traditional herbal medicines to develop a new anticancer pharmacopuncture therapy.

Kelussia odoratissima potentiates cytotoxic effects of radiation in HeLa cancer cell line.

OBJECTIVE: Cervical cancer is the second most common cause of death from cancer in women throughout the world. The aim of this study was to evaluate the cytotoxic activity of Kelussia odoratissima (K. odoratissima) extract associated with radiotherapy in cervical cancer cells (HeLa cell line). MATERIALS AND METHODS: Different concentration of the extract (25-500µg/ml) was tested in HeLa cell lines. Cell cytotoxicity of the extract and the effects of the extract on radiation (2Gy/min)-induced damages were assessed by MTT assay. Apoptosis was assessed using flow cytometric analysis. RESULT: K. odoratissima decreased cell viability in HeLa cell line in a concentration and time-dependent manner. When compared to the control, K. odoratissima induced a sub-G1 peak in the flow cytometry histogram of treated cells, indicating that apoptotic cell death is involved in K. odoratissima-induced toxicity. It was also shown that K. odoratissima sensitizes cells to radiation-induced toxicity. CONCLUSION: Our result showed the extract increased the radiation effect. This observation may be related to the presence of active compounds such as phthalides and ferulic acid.

Kelussia odoratissima Mozaff. activates intrinsic pathway of apoptosis in breast cancer cells associated with S phase cell cycle arrest via involvement of p21/p27 in vitro and in vivo.

BACKGROUND: The aim of this study was to evaluate the anticancer potential of Kelussia odoratissima. Several in vitro and in vivo biological assays were applied to explore the direct effect of an extract and bioactive compound of this plant against breast cancer cells and its possible mechanism of action. MATERIALS AND METHODS: K. odoratissima methanol extract (KME) was prepared, and MTT assay was used to evaluate the cytotoxicity. To identify the cytotoxic compound, a bioassay-guided investigation was performed on methanol extract. 8-Hydroxy-ar-turmerone was isolated as a bioactive compound. In vivo study was performed in the breast cancer rat model. LA7 cell line was used to induce the breast tumor. Histopathological and expression changes of PCNA, Bcl-2, Bax, p27 and p21 and caspase-3 were examined. The induction of apoptosis was tested using Annexin V-fluorescein isothiocyanate (FITC) assay. To confirm the intrinsic pathway of apoptosis, caspase-7 and caspase-9 assays were utilized. In addition, cell cycle arrest was evaluated. RESULTS: Our results demonstrated that K. odoratissima has an obvious effect on the arrest of proliferation of cancer cells. It induced apoptosis, transduced the cell death signals, decreased the threshold of mitochondrial membrane potential (MMP), upregulated Bax and downregulated Bcl-2. CONCLUSION: This study demonstrated that K. odoratissima exhibits antitumor activity against breast cancer cells via cell death and cell cycle arrest.

Impact of Kelussia odoratissima Mozaffarian lipid profile and fasting blood sugar in hyperlipidemia patients.

Introduction: Hyperlipidemia is associated with increased risk of cardiovascular disease. Each type of medication works differently and has different types of side effects. Flavonoids are a group of polyphenolic compounds with antioxidant properties that help reducing the cardiovascular risk factors. Kelussia odoratissima is a flavonoid containing plant. Objectives: The aim of this study was investigating the effect of this herb on lipid and glucose profile in hyperlipidemia patients. PATIENTS AND METHODS: This study performed on 61 hyperlipidemia patients. They assigned in control and intervention groups. The control group received 40 mg/day of lovastatin and intervention group received 40 mg/day of Lovastatin plus 2 g/day powder of Kelussia odoratissima. Before, two weeks and 1 month after the beginning of the study, cholesterol, triglyceride (TG), low density lipoprotein cholesterol (LDL-C), very low density lipoprotein cholesterol (VLDL-C), high density lipoprotein cholesterol (HDL-C), and fasting blood sugar (FBS) were measured. Data were analyzed by variance analysis with repeated measures, chi-square and t tests. Results: The reduction rate of cholesterol, TG, VLDL-C and LDL-C was similar in the control and intervention groups. The HDL-C rate was higher in intervention group compared to control group (P < 0.05). The mean LDL/HDL ratio decreased during the study (P < 0.001); however, there was not any significant difference between the two groups (P > 0.05). The mean of FBS did not change between and within groups (P > 0.05). Conclusion:Kelussia odoratissima did not have a desirable effect on serum lipid profile and FBS in hyperlipidemic patients that use lovastatin, but is able to increase HDL-C significantly.

Kelussia odoratissima Mozaff attenuates thromboembolic brain injury, possibly due to its Z-ligustilide content.

PRIMARY OBJECTIVE: Essential oil (EO) of Kelussia odoratissima Mozaff, whose main composition is Z-ligustilide, has been shown to have strong antioxidant and anti-inflammatory effects and potent neuroprotective properties. RESEARCH DESIGN: This study examined whether or not the EO could ameliorate brain damage and behavioural dysfunction in a thromboembolic model of stroke in rats and compare its effects to that of the purified Z-ligustilide. METHODS AND PROCEDURES: Stroke was induced in rats by middle cerebral artery occlusion using an autologous pre-formed clot. EO (10 mg kg(-1) and 45 mg kg(-1)) and Z-ligustilide (20 mg kg(-1)) were injected intraperitoneally 1 h prior to embolization. Behavioural scores, infarct size and brain oedema, as well as the level of tumour necrosis factor-alpha (TNF-α), malondialdehyde, glutathione, catalase and superoxide dismutase activity were determined in the ipsilateral cortex 24 hours following stroke induction. MAIN OUTCOMES AND RESULTS: EO (45 mg kg(-1)), statistically similar to Z-ligustilide (20 mg kg(-1)), curtailed brain infarction and oedema, improved behavioural scores and prevented enhanced oxidative stress and TNF-α level in the ischaemic brain tissues. CONCLUSIONS: The findings provide the first evidence of effectiveness of the extract in a thromboembolic model of stroke, whose action can be mediated, at least in part, by the antioxidative and anti-inflammatory mechanisms.

Antihypertensive and antioxidant effects of hydroalcoholic extract from the aerial parts of Kelussia odoratissima Mozaff. in dexamethasone-induced hypertensive rats.

BACKGROUND: Kelussia odoratissima Mozaff. is a monotypic endemic plant of Apiaceae growing wild in Iran. The aerial parts of this plant are used for treatment of hypertension, ulcer, and inflammatory conditions in folk medicine. In this study, the effects of hydroalcoholic extract of the aerial parts of K. odoratissima were evaluated in dexamethasone (Dex)-induced hypertension in male Wistar rats. MATERIALS AND METHODS: For induction of hypertension, Dex (30 µg/kg/day) was administered subcutaneously for 14 days. In a prevention study, rats received oral K. odoratissima extract (100, 200, and 400 mg/kg) from 4 days before Dex administration and during the test period (days 1-18). In a reversal study, K. odoratissima extract was administered orally from day 8 to 14. Systolic blood pressure (SBP) was evaluated using tail-cuff method. The hydrogen peroxide (H2O2) concentration and ferric-reducing antioxidant power (FRAP) were measured in plasma samples. RESULTS: Administrations of Dex significantly induced an increase in SBP and in plasma H2O2 and a decrease in body and thymus weights, and in FRAP value (P < 0.001). K. odoratissima extract dose-dependently prevented and reversed hypertension (P < 0.001). It also prevented and reduced the plasma H2O2 concentration and prevented the body weight loss upon Dex administration at all doses (100-400 mg/kg, P < 0.001) but failed to improve FRAP value. CONCLUSIONS: These results suggest antihypertensive and antioxidant effects of K. odoratissima extract in Dex-induced hypertension. Further studies are needed to elucidate the exact mechanism of the antihypertensive effect of this herbal medicine.

Kelussia odoratissima Mozzaf - a promising medicinal herb to prevent pulmonary hypertension in broiler chickens reared at high altitude.

ETHNOPHARMACOLOGICAL RELEVANCE: Kelussia odoratissima Mozzaf, formerly Apium odoratissima, is a plant locally called "Karafs", found in central Zagros region of Iran. Leaves and stems of the plant are traditionally used in the treatment of hypertension and inflammation. Lowering blood pressure effects of Kelussia odoratissima Mozzaf (wild celery) was evaluated in preventing pulmonary hypertension syndrome (PHS) in broiler chickens reared at high altitude (2,100 m). MATERIALS AND METHODS: A total number of 208 day-old male broilers (Ross 308) were randomly assigned to four treatments including different levels of Kelussia odoratissima Mozzaf (0%, 0.25%, 0.5%, and 0.75%) in a 42-day trial. RESULTS: Body weight gain and feed:gain responses significantly (P<0.05) improved when Kelussia odoratissima Mozzaf was included in broiler diets at 0.75% in the growing stage and throughout the trial. Over-expression of inducible nitric oxide (NO) synthase in the heart was observed in chickens fed Kelussia odoratissima Mozzaf. Birds received Kelussia odoratissima Mozzaf at 0.5% and 0.75% had significantly (P<0.05) higher circulatory concentrations of NO though significantly (P<0.05) lower serum malondialdehyde concentration, hematocrit and heterophil to lymphocyte ratio when compared to the birds fed the control diet. Feeding Kelussia odoratissima Mozzaf at 0.5% and 0.75% prevented from right ventricular hypertrophy and led to a significant decline in mortality from PHS. CONCLUSION: It can be concluded that Kelussia odoratissima Mozzaf is a promising medicinal herb to prevent PHS in broiler chickens by improving blood pressure and antioxidant responses.

Isolation and Identification of Ferulic Acid From Aerial Parts of Kelussia odoratissima Mozaff.

BACKGROUND: Kelussia odoratissima Mozaff. is one of the newest genera of Umbelliferae which is represented by only one species. This sweet-smelling, self-growing monotypic medicinal plant is endemic to a restricted area in west of Iran and is locally called Karafse-koohi. The aerial parts of the plant are commonly used as a popular garnish and a sedative medicinal plant. There are several reports concerning antioxidant, anti-inflammatory, anxiolytic and hypolipidemic activities of aerial parts of K. odoratissima. OBJECTIVES: The current research aimed to evaluate some phenolic contents of the plant for the first time .It is confirmed that secondary metabolites and especially phenolic compounds have important role in the biological activities of the plant. Available information indicates that phenolic contents of K. odoratissima have not been the subject of any investigation. MATERIAL AND METHODS: Aerial parts of K. odoratissima were extracted with acetone by maceration method. Normal and reversed phase vacuum liquid chromatography used to fractionate the extract. (1)H-NMR, (13)CNMR, EI-Mass and IR spectra were used to elucidate isolated compound. RESULTS: The phenolic acid isolated compound was identified as 4-hydroxy-3-methoxycinnamic acid (ferulic acid). CONCLUSIONS: Compared with previous reported antioxidant and anti-inflammatory properties of ferulic acid, a chemical-biological relation can be postulated.

Chemical composition of the essential oil from kelussia odoratissima mozaff. and the evaluation of its sedative and anxiolytic effects in mice

OBJECTIVE: The present study aimed to evaluate the sedative and anxiolytic effects of the essential oils and hydroalcoholic extract of Kelussia odoratissima Mozaff. (K. odoratissima) in mice by utilizing an elevated plus maze. The chemical composition of its essential oil was also determined. METHODS: The hydroalcoholic extract or essential oil fraction from this plant were administered intraperitoneally to male mice at various doses 30 min before testing. The anxiolytic and sedative effects were determined by an elevated plus maze and locomotor activity tests, respectively. RESULTS: According to the results, none of the administered doses of hydroalcoholic extract or essential oil fraction of K. odoratissima changed the percentage of the time spent or number of entries into the open arms of the elevated plus maze. In contrast, the cumulative spontaneous locomotor activity of mice treated with the essential oil or hydroalcoholic extract was significantly decreased. Chemical analysis of the essential oil by Gas chromatography-mass spectromentry (GC-MS) showed that 3-butylidene-4,5-dihydrophthalide (85.9 percent) was the major component. CONCLUSION: These data confirm the sedative properties of K. odoratissima, yet there were no profound anxiolytic effects observed.