In Vitro Assessment of Myorelaxant and Antispasmodic Effects of Stigmas, Tepals, and Leaves Hydroethanolic Extracts of Crocus sativus

The valorization of Crocus sativus focuses mainly on the plant’s stigma because it is one of the most expensive and valuable spices in the world and has great value in the food, cosmetics, and pharmacological industries. Due to this high stigma value, the other parts of the plant are considered as by-products; our study aimed to evaluate the myorelaxant and antispasmodic activities of the byproducts (tepals and leaves) to compare them with the stigma of the plant. To investigate the myorelaxant and antispasmodic activities of the Crocus sativus on isolated rabbit and rat jejunum, we used an in vitro technic with an organ bath and an isotonic transducer. Our results showed that the STG (hydroethanolic extract of stigmas) and LV (hydroethanolic extract of leaves) had a moderate myorelaxant effect with IC50 = 6.61 ± 1.5 and 5.08 ± 0.45 mg/ml, respectively. TPL (hydroethanolic extract of tepals) had a significant ( p ˂ 0.001 ) inhibitory effect on the amplitude of the rabbit jejunum basic contractions with an IC50 = 1.36 ± 0.15 mg/ml. TPL also caused a significant ( p ˂ 0.001 ) antispasmodic activity depending on the dose of the contraction induced by CCh (10−6 M) and KCl (25 mM). The antispasmodic effect of the TPL is slightly altered in the presence of nifedipine by a percentage of 26.8%. This difference is statistically significant ( p ˂ 0.01 ). Therefore, the extract could induce the inhibitory effect on L-type voltage-dependent Ca2+ channels, but not on the guanylate cyclase and nitric oxide pathways. That confirmed that the TPL has a comparable effect to the verapamil. The HPLC-DAD analysis of various parts of C. sativus shows that the three extracts contain the kaempferol flavanol compound, the STG also was revealed to be rich in carotenoids crocin, and these isomers are trans and cis-crocin, safranal, and picrocrocin. In contrast, the TPL revealed the presence of isorhamnetin and quercetin, but the LV was rich in hesperidin and mangiferin. In conclusion, this study supports the traditional use of this plant to treat digestive problems and will allow us to explore future possibilities for treating bowel spasms using natural molecules derived from saffron.

[1]  A. Asehraou,et al.  Crocus sativus L. (saffron): a cocktail of bioactive molecules as a biostimulant by influencing plant growth, the polyphenol and ascorbic acid content of eggplant fruit , 2022, Scientific African.

[2]  A. Bennani,et al.  Crocus sativus L. Stigmas, Tepals, and Leaves Ameliorate Gentamicin-Induced Renal Toxicity: A Biochemical and Histopathological Study , 2022, Evidence-based complementary and alternative medicine : eCAM.

[3]  S. Akhtar,et al.  Possible Mechanisms Underlying the Antispasmodic, Bronchodilator, and Antidiarrheal Activities of Polarity–Based Extracts of Cucumis sativus L. Seeds in In Silico, In Vitro, and In Vivo Studies , 2022, Pharmaceuticals.

[4]  A. Asehraou,et al.  Antioxidant Activity, Metal Chelating Ability and DNA Protective Effect of the Hydroethanolic Extracts of Crocus sativus Stigmas, Tepals and Leaves , 2022, Antioxidants.

[5]  J. Chen,et al.  The Antispasmodic Effect of Warionia saharae Essential Oil in Experimental Models and its Mechanism of Action. , 2022, Frontiers in bioscience.

[6]  J. Simal-Gándara,et al.  Anti-Depressant Properties of Crocin Molecules in Saffron , 2022, Molecules.

[7]  A. Asehraou,et al.  Protective effect of Crocus sativus stamens extract on gentamicin-induced nephrotoxicity and oxidative damage in rat kidney , 2022, Journal of Experimental Biology and Agricultural Sciences.

[8]  A. Ziyyat,et al.  Myorelaxant and antispasmodic effect of an aqueous extract of Artemisia campestris L. via calcium channel blocking and anticholinergic pathways , 2021, Journal of smooth muscle research = Nihon Heikatsukin Gakkai kikanshi.

[9]  M. Aziz,et al.  Antispasmodic activity of Warionia saharae Benthem ex Benth. & Coss. on the rabbit and rat jejunums , 2021, Journal of Pharmacy & Pharmacognosy Research.

[10]  A. Attaranzadeh,et al.  Crocus sativus (saffron) petals extract and its active ingredient, anthocyanin improves ovarian dysfunction, regulation of inflammatory genes and antioxidant factors in testosterone-induced PCOS mice. , 2021, Journal of ethnopharmacology.

[11]  Antidiarrheal Activity of Aqueous Extract of Artemisia campestris L. subsp. Glutinosa , 2021, Tropical Journal of Natural Product Research.

[12]  A. Asehraou,et al.  Hepatoprotective effects of hydroethanolic extracts of Crocus sativus tepals, stigmas and leaves on carbon tetrachloride induced acute liver injury in rats , 2021, Physiology and Pharmacology.

[13]  L. Ivanauskas,et al.  Characterization of Phytochemical Components of Crocus sativus Leaves: A New Attractive By-Product , 2021, Scientia Pharmaceutica.

[14]  S. Zara,et al.  Antimicrobial Activity and Chemical Characterization of a Non-Polar Extract of Saffron Stamens in Food Matrix , 2021, Foods.

[15]  Patrycja Pawłuszewicz,et al.  Quercetin relaxes human gastric smooth muscles directly through ATP‐sensitive potassium channels and not depending on the nitric oxide pathway , 2021, Neurogastroenterology and motility : the official journal of the European Gastrointestinal Motility Society.

[16]  A. Greco,et al.  Crocus sativus L. (Saffron) in Alzheimer's disease treatment: bioactive effects on cognitive impairment. , 2021, Current neuropharmacology.

[17]  H. Sadraei,et al.  Relaxant effect of quercetin on rabbit isolated bladder smooth muscles contractions , 2020 .

[18]  A. Asehraou,et al.  Saffron extract stimulates growth, improves the antioxidant components of Solanum lycopersicum L., and has an antifungal effect , 2019 .

[19]  Rus,et al.  Antispasmodic Effect of Essential Oils and Their Constituents: A Review , 2019, Molecules.

[20]  A. Asehraou,et al.  Antidiabetic effect of hydroethanolic extract of Crocus sativus stigmas, tepals and leaves in streptozotocin-induced diabetic rats , 2019 .

[21]  K. Sanders.  Spontaneous Electrical Activity and Rhythmicity in Gastrointestinal Smooth Muscles. , 2019, Advances in experimental medicine and biology.

[22]  P. Pittia,et al.  Crocins pattern in saffron detected by UHPLC-MS/MS as marker of quality, process and traceability. , 2018, Food chemistry.

[23]  B. Razavi,et al.  Saffron Induced Relaxation in Isolated Rat Aorta via Endothelium Dependent and Independent Mechanisms , 2018, Iranian journal of pharmaceutical research.

[24]  A. Ziyyat,et al.  Antispasmodic and Myorelaxant Activity of Organic Fractions from Origanum majorana L. on Intestinal Smooth Muscle of Rodents , 2018 .

[25]  G. Alonso,et al.  Effect of drying and storage process on Moroccan saffron quality , 2018 .

[26]  M. Elyoubi,et al.  Anti-diabetic effect of aqueous extract Crocus sativus L. in tartrazine induced diabetic male rats , 2017 .

[27]  M. Sarwat,et al.  Mechanism behind the anti-tumour potential of saffron (Crocus sativus L.): The molecular perspective. , 2017, Critical reviews in oncology/hematology.

[28]  T. Mouas,et al.  Ethnobotanical investigations on plants used in folk medicine in the regions of Constantine and Mila (North-East of Algeria). , 2016, Journal of ethnopharmacology.

[29]  M. Tsimidou,et al.  Special Issue “Saffron (Crocus sativus, L.): Omics and Other Techniques in Authenticity, Quality, and Bioactivity Studies” , 2016, Molecules.

[30]  S. Shekarforoush,et al.  The Effects of Aqueous and Alcoholic Saffron (Crocus sativus) Tepal Extracts on Quality and Shelf-Life of Pacific White Shrimp (Litopeneous vannamei) During Iced Storage , 2016 .

[31]  C. Pizza,et al.  Antioxidant activity, cytotoxic activity and metabolic profiling of juices obtained from saffron (Crocus sativus L.) floral by-products. , 2016, Food chemistry.

[32]  Manuel Carmona,et al.  Determination of saffron quality by high-performance liquid chromatography. , 2014, Journal of agricultural and food chemistry.

[33]  E. Mostafa,et al.  Ethnobotanical survey of medicinal plants used by people in Oriental Morocco to manage various ailments. , 2014, Journal of ethnopharmacology.

[34]  M. Martins,et al.  Mangiferin Prevents Guinea Pig Tracheal Contraction via Activation of the Nitric Oxide-Cyclic GMP Pathway , 2013, PloS one.

[35]  G. Alonso,et al.  A contribution to nutritional studies on Crocus sativus flowers and their value as food , 2013 .

[36]  S. Saeidnia,et al.  An overview on saffron, phytochemicals, and medicinal properties , 2013, Pharmacognosy reviews.

[37]  A. González-Coloma,et al.  Agricultural residues as a source of bioactive natural products , 2012, Phytochemistry Reviews.

[38]  G. Alonso,et al.  Increasing the applications of Crocus sativus flowers as natural antioxidants. , 2012, Journal of food science.

[39]  A. Ziyyat,et al.  Antispasmodic Effect of Aqueous Extract of Rubia tinctorum L . Roots on Rodents , 2012 .

[40]  Manuel Carmona,et al.  Geographical origin differentiation of saffron spice (Crocus sativus L. stigmas) - Preliminary investigation using chemical and multi-element (H, C, N) stable isotope analysis. , 2011, Food chemistry.

[41]  S. Z. Bathaie,et al.  HISTORICAL USES OF SAFFRON: IDENTIFYING POTENTIAL NEW AVENUES FOR MODERN RESEARCH , 2011 .

[42]  M. Boskabady,et al.  Effect of Crocus sativus ( Saffron ) on Muscarinic Receptors of Guinea Pig Tracheal Chains , 2010 .

[43]  M. Boskabady,et al.  Stimulatory effect of Crocus sativus (saffron) on beta2-adrenoceptors of guinea pig tracheal chains. , 2008, Phytomedicine : international journal of phytotherapy and phytopharmacology.

[44]  M. Zadkarami,et al.  Spasmolytic effect of Petroselinum crispum (Parsley) on rat's ileum at different calcium chloride concentrations. , 2007, Pakistan journal of biological sciences : PJBS.

[45]  Mohammad Reza Aslani,et al.  Relaxant effect of Crocus sativus (saffron) on guinea‐pig tracheal chains and its possible mechanisms , 2006, The Journal of pharmacy and pharmacology.

[46]  W. Gerthoffer.  Signal-transduction pathways that regulate visceral smooth muscle function. III. Coupling of muscarinic receptors to signaling kinases and effector proteins in gastrointestinal smooth muscles. , 2005, American journal of physiology. Gastrointestinal and liver physiology.

[47]  A. Castillo-Villanueva,et al.  Chemical Composition of Saffron (Crocus sativus L.) from Four Countries , 2004 .

[48]  J. Bornstein,et al.  Enteric motor and interneuronal circuits controlling motility , 2004, Neurogastroenterology and motility : the official journal of the European Gastrointestinal Motility Society.

[49]  M. Fatehi,et al.  Effects of Crocus sativus petals' extract on rat blood pressure and on responses induced by electrical field stimulation in the rat isolated vas deferens and guinea-pig ileum. , 2003, Journal of ethnopharmacology.

[50]  D. Criddle,et al.  Relaxant effects of the essential oil of Ocimum gratissimum on isolated ileum of the guinea pig. , 2002, Journal of ethnopharmacology.

[51]  T. Ozawa,et al.  Modulatory effect of l-NAME, a specific nitric oxide synthase (NOS) inhibitor, on stress-induced changes in plasma adrenocorticotropic hormone (ACTH) and corticosterone levels in rats: physiological significance of stress-induced NOS activation in hypothalamic–pituitary–adrenal axis , 1997, Brain Research.

[52]  R. Eglen,et al.  Characterization of the interaction between muscarinic M2 receptors and β‐adrenoceptor subtypes in guinea‐pig isolated ileum , 1995 .

[53]  A. Izzo,et al.  Effects of quercetin on the gastrointestinal tract in rats and mice , 1994 .

[54]  M. Meckes,et al.  Calcium-antagonist effect of quercetin and its relation with the spasmolytic properties of Psidium guajava L. , 1994, Archives of medical research.

[55]  M. Abou-zaid,et al.  Quercetin glycosides in Psidium guajava L. leaves and determination of a spasmolytic principle. , 1994, Archives of medical research.

[56]  H. Karaki,et al.  Calcium channels in smooth muscle. , 1984, Gastroenterology.

[57]  A. Fleckenstein.  Specific pharmacology of calcium in myocardium, cardiac pacemakers, and vascular smooth muscle. , 1977, Annual review of pharmacology and toxicology.

[58]  E. Bülbring,et al.  The inhibitory action of adrenaline on intestinal smooth muscle in relation to its action on phosphorylase activity , 1961, The Journal of physiology.