Saffron Extract Research Articles

Effective isolation protocol for secondary metabolites from Saffron: Semi-preparative scale preparation of crocin-1 and trans-crocetin

Extracts from saffron, the dried stigmata from Crocus sativus L. are recognized as valuable tools for pharmaceutical development in neuroprotection and antidepressive therapy. One major lead compound is crocin-1 (1), which gets metabolized to the C20-dicarboxylic acid trans-crocetin (2) being responsible for potential NMDA-antagonistic effects in the central nervous system. Neither crocin-1 nor crocetin are commercially available in sufficient quality and to a reasonable price. The following protocol describes effective methods to obtain both compounds from an EtOH–water extract (2:8) in good yields (about 43% related to the starting material). Crocin-1 (purity>90%) can be obtained from the extract by means of partition chromatography (FCPC) in a single run without fractionation of the mobile phase by using only the stationary phase and in yields of about 48%, related to the saffron extract. Trans-crocetin can be obtained from the EtOH–water extract by enzymatic deglycosylation of crocins using commercially available cheap glycosidase mixtures as e.g. Röhm Enzyme® or RohamentCL®. Further polishing can be achieved by flash chromatography on MCI® stationary phase with yields between 6 and 11%.The protocols described provide effective isolation of crocin-1 and trans-crocetin reference compounds for further preclinical and analytical studies with saffron extracts.

Effect of aqueous, ethanolic and acetonitrile Crocus sativus L. extracts on stress biomarkers in male rats

In this study, antianxiety-like behavior of aqueous, ethanolic and acetonitrile Crocus sativus L. extracts have been investigated in forced-swimming stress in rats. In addition, main metabolites crocin and safranal were quantified in all extracts using HPLC. Different doses of extracts (10, 30, 60 mg/kg) were injected intraperitoneally (i.p.) in a 9-day period, meanwhile, swimming stress was performed for 15 minutes in four sessions (days 3, 5, 7 and 9). The time performing the followings was measured: immobility, swimming and struggling. Moreover, free fatty acids, glucose, corticosterone and HSP70 were measured. The outcomes demonstrated saffron decreased stress significantly by prolonging immobility and decreasing the active behavior swimming, without much effect on struggling. The extracts also showed significant reduction in levels of the stress biomarkers. With having the highest amount of safranal and the lowest amount of crocin, comparing the other extracts, acetonitrile has been identified as the most effective extract in reducing anxiety. The saffron extracts probably proved anti-stress and sedative properties, partly due to distinct proportion and synergistic impact of the active constituents. On the other hand, crocin and safranal have anti-oxidant and anti-inflammatory powers that may aid to mediate this protective central impact. Regarding these information, saffron may have the potential to be employed in clinical practice. Key words: HPLC method, antianxiety-like behavior, Crocus sativus L. (saffron), forced-swimming stress, rat, stress biomarkers.

Selenium-containing lycium barbarum polysaccharides antioxidant research

Anti-inflammatory, anti oxidant and effect of Crocus sativus (C. sativus) on Th1/Th2 balance were described previously.The preventive effects of the extract of Crocus sativus on tracheal responsiveness and plasma levels of IL-4, IFN-γ, total NO and nitrite were examined on sensitized guinea pigs.Five groups of sensitized guinea pigs to ovalbumin (OVA), were given drinking water containing three concentrations of the extract of Crocus sativus, dexamethasone (S+D) or alone (group S). Tracheal responses (TR) of control animals (group C) and sensitized guinea pigs (n=6, for each group) to methacholine, OVA and the levels of IL-4, IFN-γ, total NO and nitrite in serum were examined.The TR to both methacholine and OVA, the levels of serum IL-4, total NO and nitrite in S guinea pigs were significantly increased but that of IFN-γ and IFN-γ/IL-4 ratio (Th1/Th2 balance) were decreased compared to the controls (p<0.05 to p<0.001). In the treated animals with dexamethasone and all concentrations of the extract, TR to both methacholine and OVA, IL-4, total NO and nitrite were significantly decreased but IFN-γ and IFN-γ/IL-4 ratio increased compared to S group (p<0.05 to p<0.001). The effects of the highest concentration of the extract was greater than those of other concentrations and the effect of dexamethasone (p<0.05 to p<0.01).These results not only showed a preventive effect of C. sativus extract on tracheal responses and serum levels of inflammatory mediators in sensitized guinea pigs but also showed increased Th1/Th2 balance.

Protective effect of saffron extract against doxorubicin cardiotoxicity in isolated rabbit heart

Context: Anticancer treatments such as anthracyclines are effective; however, they induce cardiotoxicity by releasing radical oxygen species (ROS). Saffron (Crocus sativus; Iridaceae) is a widely used spice with antioxidant properties and numerous health benefits that may provide cardioprotection.Objective: To assess the effect of saffron against acute myocardium damage by anthracyclines compared with electrolysis as a free radical generating system.Materials and methods: According to the Langendorff method, we used the model of an isolated rabbit heart perfused in retrograde. In one set of experiments, ROS was generated by electrolysis of the perfused heart solution (3 mA for 30 min) in the presence and absence of saffron extracts at the optimal dose (10 μg/ml). In another set, we perfused the heart with anthracycline, i.e. 30 μM doxorubicin (Doxo) in the presence and absence of 10 μg/ml saffron extracts. We evaluated cardiodynamics, as well as biochemical and pathological parameters, to emphasize the effectiveness of the treatment with saffron extract using the optimal dose of catalase (150 IU) as a positive control.Results: ROS generated, respectively, by electrolysis and by Doxo significantly (p < 0.05) affects cardiovascular function; it decreased ventricular pressure (45.02 and 40.41%), heart rate (36.31 and 22.39%) and coronary flow (50.98 and 36.67%). Increased lipid peroxidation of the myocardium was also observed (118.22 and 56.58%), while superoxide dismutase activity decreased (48.33 and 38.70%). The myocardial architecture was altered and the intercellular spaces increased.Conclusion: Saffron perfused during electrolysis helps trap ROS and significantly improves myocardial function; however, saffron was less effective against Doxo, thus suggesting that mechanisms other than oxidative stress underlie Doxo cardiotoxicity.

The Extract of Crocus sativus and Its Constituent Safranal, Affect Serum Levels of Endothelin and Total Protein in Sensitized Guinea Pigs.

The effect of the extract of Crocus sativus and its constituent, safranal on inflammatory markers in sensitized guinea pigs was examined. Ovalbumin (OA) sensitized guinea pigs were given drinking water alone (group S), or drinking water containing three concentrations of safranal, three concentrations of extract and one concentration of dexamethasone, (n=6, for all groups) and serum levels of endotheline-1 (ET-1) and total protein (TP) were assessed. Serum levels of ET-1 and TP in group S were significantly higher than control group (P<0.01 for ET-1 and P<0.001 TP). Treatment of S animals with dexamethasone, most concentrations of the extract and safranal significantly reduced serum levels of ET-1 and TP compared to group S (P <0.01 to P <0.001). The effects of one concentration of the extract and safranal were significantly higher than dexamethasone (P <0.05 to P <0.01). A preventive effect of the extract of C. sativus and its constituent safranal on serum inflammatory markers in sensitized guinea pigs was shown.

In vitro absorption studies of saffron ingredients (tr-Crocin-1, tr-Crocetin) in the Caco-2 and blood-brain barrier model

Saffron shows various effects on the central nervous system, e.g. improvement of long-term potentiation of learning and pronounced antidepressant properties, probably caused by NMDA-receptor activity and inhibition of glutamatergic synaptic transmission. Pharmacological effects have been investigated by in vitro, ex vivo and in vivo experiments, antidepressant properties were also confirmed by preliminary clinical trials. The pharmacological potential of this drug has been well studied but there is a lack of pharmacokinetic data. Since the main compounds of hydroethanolic saffron extracts are Crocins, glycosylated C-20 carotenoids, we decided to investigate the absorption properties by in vitro experiments with Crocin-1 and the aglycon Crocetin. Investigations were carried out on Caco-2 cell line, representing the intestinal barrier and on two other in vitro models representing the blood-brain barrier. The aim of the following study was to show intestinal permeation characteristics of Crocin-1 and Crocetin by Caco-2 cells and to demonstrate central nervous system availability of Crocetin by different blood-brain barrier models. The Caco-2 system was validated by quality assurance parameters as permeability of transport markers, transepithelial electrical resistance, and laser scanning microscopy. Permeation studies indicates that Crocin-1 was poorly absorbed (Papp= 2,33 *10-7 cm/s). In contrast high permeation rates were found for the aglycon Crocetin (Papp= 2,65 *10-5 cm/s). Availability of Crocetin in the central nervous system was deduced from findings of permeation through in vitro models of blood brain barrier (Papp= 1,48 *10-6 cm/s) and blood cerebrospinal barrier (Papp= 3,75 *10-6 cm/s). Based on these data it can be assumed that the glycosylated crocines are not bioavailable after oral administration, but after intestinal deglycosylation the aglycon Crocetin should be absorbed to the systemic compartment and should also permeate the blood-brain barrier.

Crocus Sativus Stigma Extract and Its Major Constituent Crocin Possess Significant Antiproliferative Properties Against Human Prostate Cancer

In this study, we investigated the antiproliferative effects of saffron extract (SE) and its major constituent crocin on 5 different malignant and 2 nonmalignant prostate cancer cell lines. Using high performance liquid chromatography (HPLC), the purity and content of crocin were determined. All cells were incubated with different concentrations of SE or crocin for 48 h. Cell cycle and apoptosis were also evaluated. In a time- and concentration-dependent manner, both SE and crocin reduced cell proliferation in all malignant cell lines with IC50 values ranging between 0.4 and 4 mg/ml for SE and between 0.26 and 0.95 mM/ml for crocin. Nonmalignant cells were not affected. Flow cytometry profiles revealed that most cells were arrested at G0/G1 phase with a significant presence of apoptotic cells. Western blot analysis revealed that the expression of Bcl-2 was strikingly downregulated, whereas Bax was upregulated. Analysis of caspase activity indicated a caspase-dependent pathway with involvement of caspase-9 activation, suggesting an intrinsic pathway. Based on these findings, it can be concluded that both SE and crocin can inhibit cell proliferation, arrest cell cycle progression, inducing apoptosis in prostate cancer. Consequently, these agents could potentially be used as a chemopreventive as well as a chemotherapeutic agent for prostate cancer management.

Functional proteomics reveals the protective effects of saffron ethanolic extract on hepatic ischemia‐reperfusion injury

Hepatic ischemia-reperfusion (IR) injury is a common clinical problem and ROS may be a contributing factor on IR injury. The current study evaluates the potential protective effect of saffron ethanol extract (SEE) in a rat model upon hepatic IR injury. Caspases 3 and terminal deoxynucleotidyl transferase-mediated dUTP biotin nick end labeling (TUNEL) results showed increased cell death in the IR samples; reversely, minor apoptosis was detected in the SEE/IR group. Pretreatment with SEE significantly restored the content of antioxidant enzymes (SOD1 and catalase) and remarkably inhibited the intracellular ROS concentration in terms of reducing p47phox translocation. Proteome tools revealed that 20 proteins were significantly modulated in protein intensity between IR and SEE/IR groups. Particularly, SEE administration could attenuate the carbonylation level of several chaperone proteins. Network analysis suggested that saffron extract could alleviate IR-induced ER stress and protein ubiquitination, which finally lead to cell apoptosis. Taken together, SEE could reduce hepatic IR injury through modulating protein oxidation and our results might help to develop novel therapeutic strategies against ROS-caused diseases.

Selective Method of Detection and Quantification of Safranal - The Natural Benzodiazepine- Like Gaba-A Receptor Agonist

Safranal, the organic volatile compound isolated from stigmas of saffron flowers (Crocus sativus) is the constituent primarily responsible for mood enhancing activity. It has been previously reported that the protective effect of safranal in anxiety and depression is mediated by the GABA-A receptor complex. Because of the commercial interest of saffron extract (2% safranal) it is essential to describe a more sensitive and discriminating methods that can quantify safranal with less interference from other components and adulterants absorbing at similar wavelength. The results here reported show that the previously reported ISO-UV/TS 3632 – 2 method (spectrometry) has lack of specificity and selectivity for the safranal analysis. The content of safranal in saffron extract is susceptible to adulteration by adding other artificial colorants as tartrazine or other parts itself of the saffron plant. The selective HPLC method here described is suitable for the detection of native safranal, not native colorants or synthetic molecules. Moreover this robust repetitive validated method, can depict the singular hplc profile of Iridafran™ saffron extract. The introduction of a photodiode array detector (DAD) let us the detection and quantification in one single run of safranal (310nm), crocin (440nm) and picrocrocin (250nm), pigments naturally content in saffron stigmas. Furthermore the method here presented is suitable for the qualitative and quantitative analysis of pigments in other botanical extracts

Protective effects of saffron (Crocus sativus) against lethal ventricular arrhythmias induced by heart reperfusion in rat: A potential anti-arrhythmic agent

Context: Saffron (Crocus sativus L.) has been used as a cuisine spice in eastern and western societies for thousands of years. In traditional medicine, saffron is recommended for the treatment of various kinds of disorders including heart palpitations.Objective: We investigated the hypothesis of the protective effect of saffron on lethal cardiac arrhythmias induced by heart ischemia-reperfusion in rat.Materials and methods: Animals were divided into a control (CTL) group that received tap water, Saf50, Saf100 and Saf200 groups that were orally treated with aqueous extracts of saffron, at dosages of 50, 100 and 200 mg/kg/day, respectively, and amiodarone (Amio) group that orally received 30 mg/kg/day for seven days. On day 8, heart ischemia-reperfusion was induced by ligation and releasing of the left anterior descending coronary artery.Results: During reperfusion, the numbers and durations of ventricular fibrillation (VF) decreased in all groups compared to the CTL group (p < 0.05). Ventricular tachycardia (VT)/VF numbers (3.2 ± 1.2), durations (4.9 ± 2.6) and also arrhythmia severity (1.9 ± 0.35) were decreased significantly in the Saf100 group versus CTL group values (18.4 ± 11.6, 52 ± 31 and 3.3 ± 0.3, respectively). The PR and QTcn intervals of ECG were significantly longer in the Saf200 group (p < 0.001 versus CTL). The other doses of saffron only significantly prolonged the QTcn interval.Conclusion: The results suggest that pretreatment with saffron, especially at the dosage of 100 mg/kg/day, attenuates the susceptibility and incidence of fatal ventricular arrhythmia during the reperfusion period in the rat. This protective effect is apparently mediated through reduction of electrical conductivity and prolonging the action potential duration.

The effect of the extract of Crocus sativus on tracheal responsiveness and plasma levels of IL-4, IFN-γ, total NO and nitrite in ovalbumin sensitized Guinea-pigs

Ethnomedical relevanceAnti-inflammatory, anti oxidant and effect of Crocus sativus (C. sativus) on Th1/Th2 balance were described previously. Aim of the studyThe preventive effects of the extract of Crocus sativus on tracheal responsiveness and plasma levels of IL-4, IFN-γ, total NO and nitrite were examined on sensitized guinea pigs. Materials and methodsFive groups of sensitized guinea pigs to ovalbumin (OVA), were given drinking water containing three concentrations of the extract of Crocus sativus, dexamethasone (S+D) or alone (group S). Tracheal responses (TR) of control animals (group C) and sensitized guinea pigs (n=6, for each group) to methacholine, OVA and the levels of IL-4, IFN-γ, total NO and nitrite in serum were examined. ResultsThe TR to both methacholine and OVA, the levels of serum IL-4, total NO and nitrite in S guinea pigs were significantly increased but that of IFN-γ and IFN-γ/IL-4 ratio (Th1/Th2 balance) were decreased compared to the controls (p<0.05 to p<0.001). In the treated animals with dexamethasone and all concentrations of the extract, TR to both methacholine and OVA, IL-4, total NO and nitrite were significantly decreased but IFN-γ and IFN-γ/IL-4 ratio increased compared to S group (p<0.05 to p<0.001). The effects of the highest concentration of the extract was greater than those of other concentrations and the effect of dexamethasone (p<0.05 to p<0.01). ConclusionsThese results not only showed a preventive effect of C. sativus extract on tracheal responses and serum levels of inflammatory mediators in sensitized guinea pigs but also showed increased Th1/Th2 balance.

Cytotoxic effect of four herbal medicines on gastric cancer (AGS) cell line

Gastric cancer (GC) is the fourth most common cancer and second leading cause of cancer-related death worldwide. Therefore, discovery of novel anti-cancer herbal drugs is of importance. Herbal medication is now being used for treatment of various diseases, including cancer in many countries. In this study, the cytotoxic effect of traditional herbal medicines (Aloe vera, Ginger, Ziziphora and Saffron extracts) was investigated on gastric AGS cell line. MTT assay was performed for cytotoxic effect of these herbal medicines. The most cytotoxic effects of Ginger, Ziziphora and Saffron extracts on AGS cell line were 47, 88 and 67%, respectively, compared to control group. According to our data, among these herbal extracts, Ziziphora has the highest cytotoxic effect on AGS cell line. Ziziphora extract seems to be a good candidate as an anti-cancer agent against GC. To clarify the effective molecules and their mechanisms, further studies are undertaken on animal models and humans.

Effective isolation protocol of saffron ingredients: Semi-preparative scale preparation of crocin-1 and trans-crocetin

Extracts from saffron, the dried stigmatas from Crocus sativus L., are recognized as valuable tools for pharmaceutical development for neuroprotection and antidepressive therapy. One major lead compound is crocin-1, which gets metabolized in vivo by deglucosylation to the C20-dicarboxylic acid trans-crocetin. Trans-crocetin is responsible for potential NMDA-antagonistic effects within central nervous system. Neither crocin-1 nor crocetin are commercially available in sufficient quality and to a reasonable price. The following protocol describes simple methods to obtain both compounds from an EtOH-water extract (2:8) of saffron.

Study of the effect of alcoholic extract of saffron on lymphocyte changes in lavage liquid in asthmatic rat

Study of the effect of alcoholic extract of saffron on lymphocyte changes in lavage liquid in asthmatic rat

Saffron Reduced Toxic Effects of its Constituent, Safranal, in Acute and Subacute Toxicities in Rats

Saffron and its constituents are widely used around the world as a spice and medicinal plant. Different constituents in medicinal herbs are thought to have the potential to induce useful and/or adverse effects. So, efforts have been made to find the best and most valuable tools to reduce their adverse effects. According to Iranian traditional medicine (ITM), it is believed that administration of whole herbs exhibits more activity and fewer side effects than isolated constituents. Since toxicological studies have indicated that safranal is more toxic than other active components in saffron stigma, thus this study was undertaken to evaluate the effect of co-administration of saffron extract and safranal in acute and sub-acute toxicities in rats. In acute toxicity, rats received safranal (1.2 mL/kg, IP) plus saffron aqueous extract (25-100 mg/kg, IP). One and four days after the treatment, percentage of mortality was assessed. In subacute toxicity, rats were randomly divided into six groups. Group 1) safranal (0.2 mL/kg, IP), Groups 2, 3 and 4) safranal plus saffron aqueous extract (5, 10 and 20 mg/kg, IP) Groups 5 and 6) Paraffin and normal saline, as solvents of safranal and saffron aqueous extract, respectively. Treatments were continued for 21 days. For sub-acute toxicity, the percentages of lethality as well as some biochemical parameters were evaluated. Our results showed that four days co-treatment of safranal and saffron significantly reduced mortality, so that the effect was more obvious in lower doses. Sub-acute toxicity studies showed that saffron could increase survival in rats so that no mortality was observed at dose of 10 mg/kg. Our data also indicated that the levels of triglyceride, BUN and ALT significantly increased after sub-acute interaperitoneal (IP) administration of safranal (0.2 mL/kg/day) and co-treatment of saffron aqueous extract (5 and 10 mg/kg) plus safranal significantly improved all toxic effects of safranal on biochemical parameters. The co-administration of saffron aqueous extract and safranal reduced toxic effects of safranal in acute and sub-acute toxicities.

SAFFRON EXTRACTS AS ENVIRONMENTALLY SAFE CORROSION INHIBITOR FOR ALUMINIUM DISSOLUTION IN 2M HCl SOLUTION

The inhibitive effect of aqueous extract of Saffron leaves toward the corrosion of aluminium in 2 M HCl solution was investigated. The inhibition efficiency was measured using weight loss and electrochemical polarization techniques. It was found that the addition of the extract reduces the corrosion rate of aluminium. The results indicate that the extracts functioned as a good inhibitor and inhibition efficiency increased with extracts concentration. Polarization curves indicated that the plant extracts behave as cathodic-type inhibitors. Surface analysis using SEM was carried out to establish the corrosion inhibitive property of this plant extract in 2 M HCl solution. The inhibitive effect of the tested extract was discussed in view of adsorption of its components on the aluminium surface. The adsorption of the extract components on the aluminium surface follows Temkin adsorption isotherm.

Cardioprotective effect of saffron extract and safranal in isoproterenol-induced myocardial infarction in wistar rats.

This study was designed to evaluate the cardioprotective effect of Crocus sativus L. (saffron) aqueous extract and safranal, the major constituent of the essential oil of saffron, on lipid peroxidation, biochemical parameters and histopathological findings in isoproterenol (ISO)-induced myocardial infarction in Wistar rats. The saffron extract (20, 40, 80 and 160 mg/kg/day IP) or control were administered for 9 days along with ISO (85 mg/kg, SC, at 24 hr interval) on 8th and 9th day in rats. Activities of creatine kinase-muscle, brain (CK-MB) and lactate dehydrogenase (LDH) were measured using standard commercial kits. The level of malondialdehyde in heart tissue was estimated with thiobarbituric acid reactive species test. For histopathological examination, hematoxylin and eosin (H&E) staining was used. ISO administration induced a statistically significant increase (P< 0.001) in serum LDH and CK-MB and a significant increase (P< 0.001) in the levels of thiobarbituric acid reactive substances (TBARs) in the heart as compared to vehicle control rats. Saffron pretreatment (20, 40, 80 and 160 mg/kg IP) or safranal pretreatment (0.025, 0.050, 0.075 ml/kg IP) for 8 days, significantly decreased (P< 0.001) the serum LDH and CK-MB and myocardial lipid peroxidation as compared to ISO- induced rats. Histological findings of the heart sections confirmed myocardial injury with ISO administration and preserved nearly normal tissue architecture with saffron or safranal pretreatment. Saffron and safranal may have cardioprotective effect in ISO-induced myocardial infarction through modulation of oxidative stress in such a way that they maintain the redox status of the cell.

Aqueous Extract of Saffron (&amp;lt;i&amp;gt;Crocus sativus&amp;lt;/i&amp;gt;) Increases Brain Dopamine and Glutamate Concentrations in Rats

Recent studies involving human and animal models have identified that saffron helps in the improvement of depression. Antidepressants are known to function in part by increasing brain serotonin, norepinephrine and dopamine concentrations. Therefore, to identify the cellular and molecular mechanism(s) underlying this property of saffron, we measured changes in rat brain dopamine, serotonin, norepinephrine and glutamate concentrations after administration of varying doses of an aqueous extract of saffron stigma. Male Wistar rats (250 ± 30 g) were administered a single dose of saffron extract (5, 25, 50, 100, 150, and 250 mg/kg, i.p.), fluoxetine (10 mg/kg, i.p.), and/or desipramine (50 mg/kg, i.p.) and were sacrificed 30 min later. Brains were removed, homogenized, and centrifuged at 4?C. The supernatant was used for subsequent neurotransmitter detection by ELISA. Our results indicated that the aqueous extract of saffron (50, 100, 150 and 250 mg/kg, i.p.) increased brain dopamine concentration in a dose-dependent manner compared with saline. In addition, the brain glutamate concentration increased in response to the highest dose of the extract (250 mg/kg, i.p.). Interestingly, the extract had no effect on brain serotonin or norepinephrine concentration. Our findings show that the aqueous extract of saffron contains an active component that can trigger production of important neurotransmitters in brain, namely, dopamine and glutamate. In addition, these results provide a cellular basis for reports concerning the antidepressant properties of saffron extract in humans and animals.

Preventive effects of hydroalcoholic extract of saffron on hematological parameters of experimental asthmatic rats.

Asthma is a chronic inflammatory disease of the respiratory airways distinguished by edema and infiltration of inflammatory immune cells. To test our hypothesis about the anti-inflammatory effect of saffron, we examined effects of Crocus sativus (C. sativus) extract as a prophylactic anti-inflammatory agent in sensitized rats. To induce experimental asthma, rats were sensitized with injection and inhalation of ovalbumin (OA). Forty male Wistar rats were divided into 5 groups (n=8 for each): control, sensitized (asthma), and sensitized and pretreated with three different concentrations of extract, 50, 100, and 200 mg/kg, 2 times a week (group asthma+50EX, group asthma+100EX, and group asthma+200EX). After 32 days, total white blood cells (WBC) counts, red blood cells (RBC), and platelet counts in blood were examined. Total WBC number and eosinophil and neutrophil percentage in blood were increased, but lymphocyte decreased in sensitized animals compared with those of control group (p<0.05 to p<0.001). We observed also elevated levels in RBC and platelet counts after sensitization in the asthma group. Pretreatment of sensitized rats in all concentrations decreased WBC count which was significant in first two concentrations (p<0.01 compared with group asthma). All concentrations of extract decreased eosinophil percentage significantly (p<0.001 compared with group asthma), however, for neutrophil percentage this improvement was not significant. Lymphocyte percentage increased in group asthma+100EX compared with group asthma (p<0.05). Moreover, in all concentrations, the extract reduced RBC and platelet count in pretreated sensitized rats compared with group asthma (p<0.01 to p<0.001). Our findings indicated that the extract of C. sativus could be useful to prevent asthma as an anti-inflammatory treatment.

Crocus sativusL. (Saffron) Stigma Aqueous Extract Induces Apoptosis in Alveolar Human Lung Cancer Cells through Caspase-Dependent Pathways Activation

Worldwide, lung cancer is the most common form of cancer. Saffron has been used in folk medicine for centuries. We investigated the potential of saffron to induce cytotoxic and apoptotic effects in lung cancer cells (A549). We also examined the caspase-dependent pathways activation of saffron-induced apoptosis against the A549 cells. A549 cells were incubated with different concentrations of saffron extract; then cell morphological changes, cell viability, and apoptosis were determined by the normal invertmicroscope, MTT assay, Annexin V and propidium iodide, and flow cytometric analysis, respectively. Activated caspases were detected by treatment of saffron in lung cancer cells using fluorescein-labeled inhibitors of polycaspases. The proliferation of the A549 cells were decreased after treatment with saffron in a dose- and time-dependent manner. The percentage of apoptotic cells increased with saffron concentrations. Saffron induced morphological changes, decreased percentage of viable cells, and induced apoptosis. Saffron could induce apoptosis in the A549 cells and activate caspase pathways. The levels of caspases involved in saffron-induced apoptosis in the A549 cells indicating caspase-dependent pathway were induced by saffron. The anticancer activity of the aqueous extract of saffron could be attributed partly to its inhibition of the cell proliferation and induction of apoptosis in cancer cells through caspase-dependent pathways activation.