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.

Anti-fatigue activity and mechanism of crocetin loaded nanoliposome in acute exercise-treated mice

Crocetin displays strong antioxidant,anti-inflammatory and anti-depression activity which is promising to relieve symptoms of fatigue.As a carotenoid,crocetin is difficult to dissolve in water and highly unstable against many environmental factors.Nanoliposome is used to encapsulate crocetin to improve its water dispersion.In the present study,the antifatigue activities and potential mechanism of crocetin loaded nanoliposome(CLN)was extensively investigated.The potential antifatigue pathway of CLN was analyzed.Furthermore,impact of CLN on the gut microbiota structure was examined which contributes to its antifatigue functions.CLN significantly increases exhaustive swimming time of fatigue mice,decreases the blood contents of lactic,blood urea nitrogen(BUN)and malondialdehyde(MDA).At the same time,CLN improves the activity of glutathione peroxidase(GSH-Px)and succinate dehydrogenase(SDH)enzyme,attenuates the oxidant stress in mice.CLN activates the adenosine monophosphate-activated kinase(AMPK)/peroxisome proliferator-activated receptor gamma coactivator-1α(PGC-1α)signaling pathway of fatigue mice,increases the mRNA expression of ATP synthase.It also increases mRNA expression of mitochondrial transcription factor A(TFAM)which promotes mitochondrial biogenesis.Additionally,CLN ameliorates the gut microbiota structure by increasing the abundance of genus such as Lactobacillus in fatigue mice.In summary,CLN exerts strong anti-fatigue properties by decreasing the oxidant stress and the contents of harmful metabolites,augmenting the production of ATP,and potentially ameliorating the gut microbiota structure.

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.

Crocetin Prevents Amyloid <i>β</i>_1-42-Induced Cell Death in Murine Hippocampal Cells

Crocetin is an aglycon of carotenoid extracted by saffron stigmas (Crocus sativus L.) and known to have a potent anti-oxidative effect. Amyliod β (Aβ), hallmark of Alzheimer’s disease, is reported to have neurotoxicity partly via oxidative stress. In this study, we investigated the effect of crocetin on hippocampal HT22 cell death induced by Aβ1-42. Furthermore, to clarify the mechanism underlying the protective effects of crocetin against Aβ1-42- induced cell death, we measured reactive oxygen species (ROS) production by CM-H2DCFDA kit assay. Crocetin at 1 -10 μM protected HT22 cells against Aβ1-42-induced neuronal cell death and decreased ROS production increased by Aβ1-42. These results that crocetin has the potent neuroprotective effect against Aβ1-42-induced cytotoxicity in hippocampal cells by attenuating oxidative stress, suggest that crocetin may provide a useful therapeutic strategy against Aβ-related disorders.

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.

Crocetin Potentiates Neurite Growth in Hippocampal Neurons and Facilitates Functional Recovery in Rats with Spinal Cord Injury

Crocetin is an ingredient of traditional Chinese medicine and has therapeutic potential in various diseases due to its pharmacological properties, such as neuroprotection, anti-oxidative stress, and anti-inflammation. These properties might benefit the treatment of spinal cord injury.In the present study, we tested the effect of crocetin on neurite growth and sensorimotor dysfunction in a rat model of spinal cord injury. We evaluated the viability of cultured hippocampal neurons with tetrazolium dye and lactate dehydrogenase assays, visualized neurites and axons with antibody staining, and monitored motor and sensorimotor functions in rats with spinal cord injury using the Basso,Beattie, and Bresnahan assay and the contact plantar placement test, respectively, and measured cytokine expression using enzyme-linked immuno-absorbent assays.We found that crocetin（1） did not alter the viability of cultured hippocampal neurons;（2） accelerated neurite growth with preference for the longest process in individual hippocampal neurons;（3） reversed the inhibition of neurite growth by chondroitin sulfate proteoglycan and Nogo A;（4） facilitated the recovery of motor and sensorimotor functions after spinal cord injury; and（5） did not inhibit pro-inflammatory responses, but restored the innervation of the descending 5-HT system in injured spinalcord. Crocetin promotes neurite growth and facilitates the recovery of motor and sensorimotor functions after spinal cord injury, likely through repairing neuronal connections.

Crocin Synthesis Mechanism in Crocus sativus

Saffron （Crocus sativus） cells can synthesize crocin, crocetin digentiobiosyl ester, in suspension cultures. The crocin family biosynthesis mechanism was studied using high pressure liquid chromatography （HPLC） to determinate the glucosyltransferase activity and to develop a method for synthesizing medicine from saffron cells. Previous studies indicated that two glucosyltransferases might be involved in the formation of crocetin glucosyl- and gentiobiosyl-esters. GTasel formed an ester bond between crocetin carboxyl groups and glucose moieties while GTase2 catalyzed the formation of glucosidic bonds with glucosyl ester groups at both ends of the molecule. These enzymes can catalyze the formation of crocetin glucosides in vitro. GTasel activity is higher during the first four days of crocin glucosides biosynthesis, but decreases after four days. The formation and accumulation of crocin increase during the first six days and stabilized on the eighth day.