Inhibitory effect of saffron on head and neck squamous cell carcinoma via targeting of ESR1 and CCND1 by its active compound crocetin

Background:Traditional Chinese medicine is promising for managing challenging and complex disorders,including cancer,and in particular,saffron is applied in treating various cancer types.However,its potential therapeutic efficacy and active components in managing squamous cell carcinoma of the head and neck(HNSCC)remain unclear yet.Methods:Using network pharmacology approaches,active ingredients of saffron,their target genes,and HNSCC-related genes were identified.Enrichment analyses were conducted for determining molecular functions and pathways enriched by genes that overlapped between the saffron target gene set and the HNSCC gene set.Among the four known active ingredients of saffron,crocetin was found to have the strongest inhibitory impact on HNSCC,based on the findings of cell viability and migration assays.Therefore,the potential target genes of crocetin in HNSCC cells were examined using molecular docking experiments and were confirmed by qPCR.Result s:Four active ingredients of saffron and 184 of their target genes were identified.Further,a total of 34 overlapping saffron-/HNSCC-associated targets related to the four active ingredients were screened,and crocetin was chosen for further investigation because it had the strongest inhibitory effect on HNSCC cells.Molecular docking experiments indicated that ESR1 and CCND1 were the target genes of crocetin.These results were confirmed through qPCR analysis,in which crocetin was found to lower the expression of the ESR1 and CCND1 genes in AMC-HN-8 and FaDu cells.Conclusion:According to our results,crocetin is a primary active anti-cancer component of saffron that may have potential in the development of novel HNSCC-treating medications.However,more thorough molecular research is necessary for confirming these results and elucidating the anti-cancer mechanism underlying saffron.

Saffron(Crocus sativus L.) stigmas as a potential natural additive to improve oxidative stability attributes of sunflower(Helianthus annuus L.) oil stored under different conditions

The aim of our study was to explore the antioxidant potential of Moroccan saffron stigmas(SS) and examine their performance as a natural antioxidant in enhancing the stability and quality properties of edible sunflower oil(SO), with a focus on understanding the critical importance of edible oil stability for predicting quality deterioration during storage. Bearing this in mind, our research was designed to compare the efficacy of three concentrations of dried SS(at 0.2%, 0.3%, and 0.6%) against tocobiol(a synthetic antioxidant) as a positive control(at 0.3% concentration). This comparison was conducted under three distinct storage conditions namely accelerated ageing(60 ℃), exposure to light at ambient temperature, and darkness, to evaluate their impact on preventing severe oxidation and extending oil shelf-life. Oxidation state evolution was evaluated through peroxide value(PV), free fatty acid(FFA), anisidine value(p-AV), K_(270)value(conjugated trienes), total oxidation index(TOTOX), iodine value(IV), and fatty acid composition(FA). Our results revealed notable differences in stability tracking parameters. Specifically, these parameters were higher in samples stored under accelerated conditions, followed by the samples stored in ambient light, while those stored in darkness showed the highest stability among the three storage conditions. Supplementation of sunflower oil with SS and tocobiol significantly enhanced its oxidation stability. Notably, SS exhibited exceptional effectiveness in stabilizing sunflower oil regardless of SS dose, with the highest efficacy observed at 0.6%. This was evidenced by the slower rate of oxidation parameters under various storage conditions, highlighting a superior antioxidant activity compared to both the non-enriched oil and tocobiol-enriched oil. Furthermore, saffron stigmas, used as a natural supplement,contributed to the preservation of polyunsaturated fatty acids, indicating its potential as a robust source of natural antioxidants in sunflower oil. These attributes position SS as a promising alternative to synthetic antioxidants, offering opportunities to enhance the nutritional quality and extend edible oil shelf-life.

Beneficial effects of saffron(Crocus sativus L.) in ocular pathologies, particularly neurodegenerative retinal diseases

Saffron(Crocus sativus L.)has been traditionally used in food preparation and as a medicinal plant.It currently has numerous therapeutic properties attributed to it,such as protection against ischemia,as well as anticonvulsant,antidepressant,anxiolytic,hypolipidemic,anti-atherogenic,anti-hypertensive,antidiabetic,and anti-cancer properties.In addition,saffron has remarkable beneficial properties,such as anti-apoptotic,anti-inflammatory and antioxidant activities,due to its main metabolites,among which crocin and crocetin stand out.Furthermore,increasing evidence underwrites the possible neuroprotective role of the main bioactive saffron constituents in neurodegenerative diseases,such as Parkinson’s and Alzheimer’s diseases,both in experimental models and in clinical studies in patients.Currently,saffron supplementation is being tested for ocular neurodegenerative pathologies,such as diabetic retinopathy,retinitis pigmentosa,age-related macular degeneration and glaucoma,among others,and shows beneficial effects.The present article provides a comprehensive and up to date report of the investigations on the beneficial effects of saffron extracts on the main neurodegenerative ocular pathologies and other ocular diseases.This review showed that saffron extracts could be considered promising therapeutic agents to help in the treatment of ocular neurodegenerative diseases.

Green synthesis of silver nanoparticles using aqueous extract of saffron(Crocus sativus L.) wastages and its antibacterial activity against six bacteria

Objective: To synthesis silver nanoparticles(Ag NPs) by using extract of saffron(Crocus sativus L.) wastages and to test their antibacterial activity against six bacteria.Methods: In this paper, the synthesis of Ag NPs using aqueous extract of saffron wastage as a green method without any chemical stabilizer and reducer is demonstrated. The synthesized Ag NPs were determined by UV–vis spectrum, high resolution transmission electron microscopy, X-ray diffraction, and Fourier transmission infrared spectroscopy analysis.Results: UV–vis spectrum showed a peak at 450 nm due to excitation of surface plasmon vibrations. Fourier transmission infrared spectroscopy showed that nanoparticles were capped with plant secondary metabolites. X-ray diffraction analysis also demonstrated that the size range of the synthesized nanoparticles was 12–20 nm. Transmission electron microscope image illustrated Ag NPs with spherical shape and an average size of15 nm. The result of antibacterial activities showed that the biosynthesized Ag NPs had an inhibiting activity against Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Shigella flexneri and Bacillus subtilis.Conclusions: The biosynthesized Ag NPs showed significant antibacterial effect against Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Shigella flexneri and Bacillus subtilis, so, it can be used in biomedical applications.

Biochemical, Antioxidant and Antineoplastic Properties of Italian Saffron (<i>Crocus sativus L</i>.)

Saffron, the most expensive spice in the world, is got by Crocus sativus L. stigmas. The production of this substance has attracted human interest, since ancient cultures, for its medicinal and culinary properties. Because of saffron high economic value, sometimes, since Middle Ages, it is adulterated with other vegetal materials, dyes or synthetic molecules. Object of this work was the study of one of the best world saffron: Civitaretenga (AQ, Central Italy) C. sativus. Taste, color and aroma of Civitaretenga spice were determined, according to international standards (ISO/Technical Specification 3632), to define its high quality. A biochemical approach was then applied to obtain a secondary metabolite profile of this product. So, crocins, total phenolic content, flavonoids and phenolic acids were detected by HPLC-DAD and spectrophotometric analysis. Moreover, in vitro antioxidant properties and in vivo antineoplastic effects, on highly metastatic murine B16-F10 melanoma cell line, were successfully revealed in Civitaretenga C. sativus extract. All these data confirmed the elevated quality of Civitaretenga saffron and its highly reducing and chemopreventive activity.

The History of Saffron in China:From Its Origin to Applications

Saffron(Stigma Croci)is an autumn-flowering perennial plant,and its use has a history of over 3500 years.Saffron has often been considered as the costliest medicinal plant,a premium spice,and the best dye with a golden yellowish color.Iran currently produces the finest quality saffron and dominates its global production(>90%).Other countries such as Australia,Canada,the USA,China,and some countries in Central Africa,produce saffron at a lower yield.In China,saffron is celebrated as“red gold”owing to the red stigmas of the flower and its price,which is comparable to the price of gold.Saffron has been one of the most attractive traditional Chinese medicine(TCM)herbs in the Zhong Guo Yao Dian(《中国药典》Chinese Pharmacopoeia)since its inclusion in the 2005 edition.The earliest use of saffron in TCM was recorded in the Ben Cao Shi Yi(《本草拾遗》Supplement to Materia Medica)written during the Tang dynasty(741 A.D.).However,saffron grown in inland China has been widely mistaken as originating from Tibet.This is because its Chinese name begins with“Xi”or“Zang,”which sounds similar to its Tibetan name(“Xi Zang”).In this review,we clarify the origin of saffron and its introduction to China and summarize its various applications.

Saffron (<i>Crocus sativus L.</i>) Inhibits Aflatoxin B₁Production by <i>Aspergillus parasiticus</i>

Aflatoxin B1 (AFB1) is a carcinogenic metabolite produced by certain Aspergillus species. The aim of the present study was to investigate the effect of saffron stigmas on A. parasiticus growth and AFB1 production in Yeast Extract Sucrose (YES) medium. AFB1 was extracted from cultures and purified with immunoaffinity columns followed by high performance liquid chromatography (HPLC) coupled to fluorescence detector (FL) analysis. Methods' recovery and limit of detection were 95.3% and 0.02 ng AFB1·ml–1 of YES respectively. Results indicated that AFB1 production in samples of YES inoculated with A. parasiticus after the addition of saffron dried stigmas (100 mg·flask–1) was significantly lower (p 1 was detected, after the 7th day of observation in cultures with saffron alone. Maximum production was observed on the 12th day (0.018 μg AFB1·flask–1) and on the 9th day (0.051 μg AFB1·flask–1) for samples of YES with the addition of saffron inoculated with A. parasiticus and samples with saffron alone (non-inoculated), respectively. In control samples (inoculated without saffron) the maximum production on the same days 12 and 9 was 75.31 μg AFB1·flask–1 and 64 μg AFB1·flask–1 respectively. Conclusively when saffron was added to YES inoculated with A. parasiticus, AFB1 production decreased by 99.9% compared to control cultures without saffron addition. This inhibition can be attributed to the antioxidant capacity of saffron constituents.

Neuroprotective properties of dietary saffron：more than just a chemical scavenger？

There is increasing evidence that consumption of saffron,a spice derived from the flower of the Crocus savitus plant,has various therapeutic effects,including protection of the central nervous system.

Aqueous Extract of Saffron (<i>Crocus sativus</i>) 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.

Mechanisms of saffron in treatment of atherosclerosis based on network pharmacology

Objective:To explore the main pathways and possible mechanisms of saffron anti-atherosclerosis(AS)using network pharmacology.Methods:The active ingredients and target proteins of the drug were obtained from the TCMSP database,and the gene names corresponding to the target proteins were queried using the Uniprot database to obtain human targets regulated by saffron.Human targets of AS were obtained from DisGeNET database and previous literature.The target gene of saffron was intersected with the target gene of AS to obtain the target gene of saffron for AS.With the help of Cytoscape 3.7.2 software,a saffron active ingredient-potential target,saffron active ingredient-AS disease target interaction network was constructed,and the core targets of saffron's role in AS disease were screened out through evaluation of network topological characteristics such as degree;The ClueGo plug-in in Cytoscape 3.7.2 software was used to analyze GO biological processes and KEGG metabolic pathways of targets.Results:With the drug-like property≥0.18 as the limiting condition and previous literature search and screening,22 compounds were obtained.A total of 106 saffron-regulated target proteins and 117 AS targets were obtained.The intersections were obtained to obtain 26 saffron targets for AS.According to the degree of drug-candidate component-AS candidate target network,the main effective components of saffron anti-AS were crocetin,isorhamnetin and quercetin.26 gene targets were analyzed by KEGG enrichment using the ClueGo plug-in in Cytoscape 3.7.2 software to obtain pathways related to saffron treatment of AS.Using the KEGG database and consulting previous literature,a schematic diagram of the related pathways of saffron anti-AS was obtained.Conclusion:The anti-AS of saffron may be mainly caused by crocetin,isorhamnetin and quercetin in saffron to exert anti-inflammatory and inhibit angiogenesis effects,thereby achieving anti-AS effect.

Survey of the History and Applications of Saffron

Fan Hong Hua(番红花saffron)is a natural product that has long been used in food and dye industries and fortreating various disorders.Saffron has a long history of applications in traditional Chinese medicine,following its introduction to China in 1979.According to the theory of traditional Chinese medicine,saffron is sweet,slightly cold in nature and enters heart and liver meridians.Moreover,this spice can invigorate blood circulation,eliminate blood stasis,cool blood and eliminate heat toxins.Saffron has been used to treat asthma,phlegm,insomnia,Alzheimer’s disease,depression,fright,shock,hemoptysis,heartburn,and pain.Among the almost 70 bioactive ingredients of saffron,major constituents of this herb include safranal,crocin,and crocetin,which are responsible for the unique taste and color of saffron.These ingredients have been investigated using modern pharmaceutical research methods as potential therapeutics to treat disorders such as Alzheimer’s disease,cardiovascular diseases,cancer,digestive system disorders,ocular issues,and COVID-19 infections.However,focus and emphasis on understanding the pharmacological effects of saffron in treating diseases are required.This article briefly reviews the history,cultivation processes,different qualities properties,and traditional and modern applications of this unique herb.

Isolation of Main Genes Involved in Flowering of Saffron （Crocus sativus L.） in Order to Study Effective Flowering Protein

Saffron （Crocus sativus L.） always is grown for using its flowers in nutrient industry, color industry and healthy compounds due to its flowers and specially stigmas. Because of its expensive flowers, surveying and recognizing on effective genes for flowering is very important and its results can help us to control rate and timing of flowering at an early stage of flowering. The gene and gene state meant Pistillata like MADS box （PIC2） were surveyed for recognizing its molecular mechanism. The molecular sequence at the genes has high similarity to members of family MADS that is a factor for controls of protein at flowering stage. PIC2 gene was studied by bioinforrnatics resources. Primers were designed for replicating the gene and DNA and RNA were extracted from saffron＇s leaves. The gene＇s eDNA was built by recopying enzyme and used such a pattern for replicating gene PIC2 at polymerase chain reactions （PCR）. Segments were replicated such 900 eDNA pair-nucleotides and a segment such 2,100 of DNA＇s pair-nucleotides. The gene codes a protein that was composed of 210 amino acids that has MADS sequence box. Analysis of protein＇s molecular structure and homological modeling of the protein indicated that it has a regular structure.

Purification of Agro Waste Saffron Using Membrane Technology-Ultrafiltration-Application to Sunscreen Cosmetic Emulsions

In this present work, ultrafiltration was applied for the purification of agro waste Greek red saffron. The agro waste saffron (before and after ultrafiltration) was used for the preparation of oil- in-water emulsions. Produced emulsions were examined for their viscosity, pH and conductivity during time. Also surface tension values of the aqueous and oily phase of the emulsions were measured. Saffron is known for its antioxidant and anti-solar properties. Anti-solar activity of saffron was evaluated by measuring the SPF value of the emulsion. UV-Vis measurements have shown an incredible increase of the absorption of saffron after ultrafiltration. The emulsions made with the purified saffron presented a better anti-solar activity and at the same time remarkably improvement of their antioxidant and antiaging properties giving a very promising alternative to the cosmetic industry.

Saffron： a potential immunological adjuvant in anti-tumor therapy

A large and increasing number of patients in the world use medicinal plants and herbs for health purposes.Especially,chemoprevention using readily available natural substances from vege.tables,fruits,herbs and spices is one of the significantly important approaches for cancer prevention in the present era.Saffron is native to Iran and now recorded in "Chinese Pharmacopoeia" as a widely used Chinese medicine with good safety.Other than several useful pharmacological effects such as anticonvulsant,antidepressant,anti-inflammation,saffron and its active components(crocin,crocetin and safranal) have been shown to induce apoptosis in several tumor cell lines and mouse tumors with beneficial properties including radical scavenging,anti-mutagenic and immuno-modulating effects.In addition,saffron was reported good potential to alleviate the toxicity ofcisplatin,including the nephrotox.icity.However,the application of these components in the clinic has been limited due poor clinical trials data.This review aimed to provide a brief overview on clinical evaluation for anti-tumor potential and current molecule mechanism of saffron based on recent literature data.

Neuroprotective effects of saffron on chronic focal cerebral ischemia through inhibiting glial scar formation in rats

OBJECTIVE To explore the neuro-protective effects of saffron(Crocus satius L.) on chronic focal cerebral ischemia in rats.METHODS SD rats were randomly divided into 6 groups:sham control group,MCAO group,edaravone group and saffron 30,100,300 mg·kg^(-1) groups.Focal cerebral ischemia was induced by middle cerebral artery occlusion(MCAO).Saffron was administered orally by once daily from 2 h to 42 d after ischemia.At 42 d after cerebral ischemia,neurological deficit score,spontaneous activity test,elevated plus maze test,marble burying test and novel objective recognition test were used to evaluate the effects of saffron on the behevioural change.Infarct volume,survival neuron density,activated astrocyte number,and the thickness of glial scar were also detected.GFAP expression and inflammatory cytokine contents in ischemic peripheral region were detected by Western blot and ELISA,separately.RESULTS Saffron(100,300 mg·kg^(-1)) improved the body weight decrease,neurological deficit and spontaneous activity.Saffron(30-300 mg · kg^(-1)) increased the traveled distance ratio and total time in open arm,decreased the buried marble number,which indicated that saffron could ameliorate anxiety-and depression-like behaviors.Saffron(100,300 mg·kg^(-1)) improved the learning and memory function,which manifested by increased discrimination ratio(DR) and discrim.ination index(DI) in T2 test.The results of toluidine blue found saffron treatment(100,300 mg · kg^(-1))decreased the infarct volume and increased the neuron density in cortex and hippocampal.The activated astrocyte number,the thickness of glial scar and GFAP expression in ischemic peripheral region decreased after saffron.Saffron(100,300 mg · kg^(-1)) decreased the contents of IL-6 and IL-1β,increased the content of IL-10 in ischemic peripheral region.CONCLUSION Saffron exerted neuro-protective effects on chronic focal cerebral ischemia,which could be related with inhibiting the activation of astrocyte and glial scar,following with the decrease of inflammatory reaction.

Analysis of volatile components in saffron from Tibet and Henan by ultrasonic-assisted solvent extraction and GC-MS

To determine the chemical constituents of volatile components in saffron from the Tibet Autonomous Region and the Henan Province of China and to compare the chemical composition difference in the saffron,the total volatile components were extracted by ultrasonic-assisted solvent extraction（USE）,using five different solvents：diethyl ether,ethanol,ethyl acetate, dichloromethane and acetone,analyzed using gas chromatography-mass spectrometry（GC-MS）and compared to the standard MS data,and their relative contents were calculated by area normalization.The results showed both that USE was an efficient and rapid method for the extraction of the volatile components from saffron and that the components extracted from the same sample using different solvents were different.Comparison of the experimental results of saffron from the Tibet Autonomous Region and the Henan Province of China showed that their volatile components were different in their chemical composition and in their relative percentages.USE/GC-MS is a simple,rapid,and effective method for the analysis of volatile oil components in saffron.

Determination of the major metal elements including heavy metals in Saffron from Tibet and Henan by ICP-AES or ICP-MS

A quick and sensitive method was developed for the determination of 19 metal elements in Saffron,a traditional Chinese medicinal herb,collected from Tibet Autonomous Region and Henan Province of China using inductively coupled plasma atomic emission spectrometry（ICP-AES）or inductively coupled plasma mass spectrometry（ICP-MS）.The determined 19 metals in Saffron include Ca,Fe,Mg,P,Sr,Al,Mn,Zn,V,Cr,Se,Co,Ni,Mo,and heavy metals As,Cu,Cd,Hg and Pb.For all the analyzed elements,the correlative coefficients of the calibration curves were no less than 0.9938.This proposed method was accurate,and the relative standard deviations of the measurements were lower than 5.25%.It can be used for the quality control of metal elements in Saffron.

Effects of Culture Conditions, Carbon Source and Regulators on Saffron Callus Growth and Crocin Accumulation in the Callus

There are many factors influencing the growth and secondary metabolites of callus and saffron callus. In this paper, the effects of culture conditions, including culture temperatures, light levels, the carbon source and its concentration, and the preserve of regulators (mainly hormones), are studied for callus cultures. All the experiments used Murashige and Skoog (MS) solid medium as the basic medium with 10 g/L agar, pH 5.75.8. Saffron callus was cultured at 20℃ in the dark, with a sucrose concentration of 45 g/L (or starchy hydrolysate concentration of 40 g/L), but 30 g/L sucrose was best for the synthesis of crocin (for starchy hydrolysate the concentration can range from 20 to 40 g/L). To promote callus growth, the best auxin was α-naphthaleneacetic acid (NAA) and the optimum ratio of NAA (mg/L) to benzylaminopurine (BA) (mg/L) was 2∶0.25. Indole-3-acetic acid (IAA) (4 mg/L), gibberellin(GA3) (2 mg/L), and uniconazole (S-07) (1.25 mg/L) increased the crocin content remarkably as analyzed by high performance liquid chromatography (HPLC). NAA (2 mg/L) promoted the growth of saffron callus but had no benefit and may inhibit crocin synthesis while S-07 (1.25 mg/L) had the opposite effect. GA3 promoted both growth and synthesis.

Clinical Evidence on the Effects of Saffron(Crocus Sativus L.)in Anxiety and Depression

Crocus sativus L.,a member of the Iridaceae family,commonly known as saffron has very much popular as both medicine and spice.Earlier saffron was used as one of the important constituents in opioid preparations for pain relief.Its uses in the treatment of mild-to-moderate depression are well documented.Depression can affect the quality of life of individuals by causing headaches,difficulty in thinking,and loss of interest.Many clinical trials have been conducted for evaluating the efficacy of saffron in depression cases.Almost fifty constituents of C.sativus L are well known.Few of the major constituents include crocin,picrocrocin,and safranal.In the present review,the main focus is on the antidepressant effects of saffron and its major constituents,in which it was found that saffron has substantial effects on depression and anxiety because of double-blind study the data can be trusted.

DEVELOPMENT OF A NOVEL HPLC METHOD FOR QUALITY CONTROL OF SAFFRON(CROCUS SATIVUS L.)

Saffron,the dried stigma of Crocus sativus L.,findsnumerous applications in TCM.Here,a novel HPLC protocol was established and applied for the analysis of saffron samples,not only from different places of origin but also from several harvest seasons.One of the main active constituents of saffron,crocin,is also contained in