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Activity of Antioxidants from Crocus Sativus L. Petals: Potential Preventive Effects Towards Cardiovascular System

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Activity of Antioxidants from Crocus Sativus L. Petals: Potential Preventive Effects Towards Cardiovascular System antioxidants Article Activity of Antioxidants from Crocus sativus L. Petals: Potential Preventive Effects towards Cardiovascular System 1, 2, 3, 4 Keti Zeka y , Pasquale Marrazzo y , Matteo Micucci * , Ketan C. Ruparelia , Randolph R. J. Arroo 4 , Guido Macchiarelli 5 , Stefania Annarita Nottola 6, Maria Adelaide Continenza 5, Alberto Chiarini 3, Cristina Angeloni 7 , Silvana Hrelia 2 and Roberta Budriesi 3 1 Department of Haematology, Cambridge Biomedical Campus, University of Cambridge, Long Road, Cambridge CB2 0PT, UK; [email protected] 2 Department for Life Quality Studies, Alma Mater Studiorum-University of Bologna, Corso d’Augusto 237, 47921 Rimini, Italy; [email protected] (P.M.); [email protected] (S.H.) 3 Nutraceutical Lab, Department of Pharmacy and Biotechnology, Alma Mater Studiorum, University of Bologna, Via Belmeloro 6, 40126 Bologna, Italy; [email protected] (A.C.); [email protected] (R.B.) 4 Leicester School of Pharmacy, Faculty of Health and Life Sciences, De Montfort University, The Gateway, Leicester LE1 9BH, UK; [email protected] (K.C.R.); [email protected] (R.R.J.A.) 5 Department of Life, Health and Environmental Sciences, University of L’Aquila, Via Vetoio, Coppito 2, 67100 L’Aquila, Italy; [email protected] (G.M.); [email protected] (M.A.C.) 6 Department of Anatomy, Histology, Forensic Medicine and Orthopaedics, La Sapienza University of Rome, 00161 Rome, Italy; [email protected] 7 School of Pharmacy, University of Camerino, Via Madonna delle Carceri 9, 62032 Camerino (MC), Italy; [email protected] * Correspondence: [email protected] These authors contributed equally to this work. y Received: 3 September 2020; Accepted: 6 November 2020; Published: 9 November 2020 Abstract: The petals of the saffron crocus (Crocus sativus L.) are considered a waste material in saffron production, but may be a sustainable source of natural biologically active substances of nutraceutical interest. The aim of this work was to study the cardiovascular effects of kaempferol and crocin extracted from saffron petals. The antiarrhythmic, inotropic, and chronotropic effects of saffron petal extract (SPE), kaempferol, and crocin were evaluated through in vitro biological assays. The antioxidant activity of kaempferol and crocin was investigated through the 20,70-dichlorodihydrofluorescein diacetate (DCFH-DA) assay using rat cardiomyoblast cell line H9c2. The MTT assay was applied to assess the effects of kaempferol and crocin on cell viability. SPE showed weak negative inotropic and chronotropic intrinsic activities but a significant intrinsic activity on smooth muscle with a potency on the ileum greater than on the aorta: EC50 = 0.66 mg/mL versus EC50 = 1.45 mg/mL. Kaempferol and crocin showed a selective negative inotropic activity. In addition, kaempferol decreased the contraction induced by KCl (80 mM) in guinea pig aortic and ileal strips, while crocin had no effect. Furthermore, following oxidative stress, both crocin and kaempferol decreased intracellular ROS formation and increased cell viability in a concentration-dependent manner. The results indicate that SPE, a by-product of saffron cultivation, may represent a good source of phytochemicals with a potential application in the prevention of cardiovascular diseases. Keywords: cardiovascular prevention; nutraceuticals; crocin; Crocus sativus L. petals; kaempferol; by-products Antioxidants 2020, 9, 1102; doi:10.3390/antiox9111102 www.mdpi.com/journal/antioxidants Antioxidants 2020, 9, 1102 2 of 18 Antioxidants 2020, 9, x FOR PEER REVIEW 2 of 19 1. Introduction 1. Introduction For centuries, plants have represented the main tools for the treatment of human diseases. In theFor modern centuries, era, plants phytochemistry have represented and pharmaceutical the main tools for approaches the treatment were of human applied diseases. to investigate In the themodern mechanisms era, phytochemistry of action and and the pharmaceutical chemical composition approaches of plants were usedapplied in folkto investigate medicine the [1,2 ]. mechanisms of action and the chemical composition of plants used in folk medicine [1,2]. The The pharmaceutical value of plant extracts may be due to the synergy of the phytochemicals present pharmaceutical value of plant extracts may be due to the synergy of the phytochemicals present in in them, even if, rarely, the affection of specific functions may be ascribed to isolated molecules. them, even if, rarely, the affection of specific functions may be ascribed to isolated molecules. Phytochemicals are non-nutrients compounds normally found in plants used as food or as components Phytochemicals are non-nutrients compounds normally found in plants used as food or as of food supplements, able to exert several biological effects which potentially result in the modulation components of food supplements, able to exert several biological effects which potentially result in of several parameters related to human health [3]. the modulation of several parameters related to human health [3]. Cardiovascular diseases (CVDs) are considered a major cause of mortality worldwide. According to Cardiovascular diseases (CVDs) are considered a major cause of mortality worldwide. the World Health Organization (WHO), 17.9 million people were estimated to die from CVDs in According to the World Health Organization (WHO), 17.9 million people were estimated to die from 2017,CVDs representing in 2017, representing 31% of all global31% of deathsall global [4]. deaths Of this [4]. mortality, Of this anmortality, estimated an 7.4estimated million 7.4 were million due to coronarywere due heart to coronary disease andheart 6.7 disease million and to stroke.6.7 million In the to age stroke. of genomics, In the age nanotechnology, of genomics, nanotechnology, and proteomics, CVDsand proteomics, continue to CVDs be a majorcontinue challenge to be a major and development challenge and of development a viable evidence-based of a viable evidence-based way to prevent theway onset to prevent of CVDs the continues onset of CVDs to attract continues the attention to attract of the researchers. attention of researchers. SeveralSeveral compounds compounds havehave beenbeen shown to exert biolog biologicalical actions actions that that may may result result in in a adecrease decrease of of cardiovascularcardiovascular diseasesdiseases [[5,6].5,6]. ForFor example, flavonoids flavonoids can can improve improve cardiovascular cardiovascular health health through through severalseveral mechanisms mechanisms including including the the antioxidant antioxidant effect, effect, the hypocholesterolemic the hypocholesterolemic and antidiabetic and antidiabetic activities, theactivities, antiatherosclerotic the antiatherosclerotic and vasorelaxant and vasorelaxant properties [7properties,8]. Similarly, [7,8]. carotenoids Similarly, carotenoids such as crocin, such lutein, as zeaxanthincrocin, lutein, exert zeaxanthin biological exert actions biological that may actions determine that may a plethora determine of cardiovascular a plethora of benefitscardiovascular [9,10]. benefitsSome [9,10]. spices and vegetal extracts exert antioxidant [9] and hypocholesterolemic effects [11,12]. SaffronSome (Crocus spices sativus and vegetalL.) is one extracts of the exert most antioxid popularant spices [9] and [13 ].hypocholesterolemic It is commonly used effects in regions [11,12]. of EuropeSaffron such (Crocus as Italy, sativus Spain, L.) is and one Greece, of the andmost in popular Asian regions spices [13]. of the It world,is commonly such as used Iran in and regions India [of14 ]. Briefly,Europe sa suchffron as is aItaly, spice Spain, derived and from Greece, the flowerand in ofAsianCrocus regions sativus of Lthe (Iridaceae). world, such Each as flowerIran and has India three stigmas,[14]. Briefly, which saffron are dried is a spice and usedderived in cookingfrom the as flower a seasoning of Crocus and sativus as a coloringL (Iridaceae). agent. Each They flower contain has a carotenoidthree stigmas, dye, which crocin, are which dried gives and foodused a in rich cooking golden-yellow as a seasoning hue [7 and,15,16 as]. a Sa coloringffron consumption agent. They is showncontain to a correlate carotenoid with dye, higher crocin, High which Density gives Lipoproteins food a rich (HDL) golden-yellow and lower triglycerideshue [7,15,16]. levels Saffron [17 ]. Moreover,consumption administration is shown ofto 15correlate mg of sa withffron capsuleshigher High to diabetic Density patients Lipoproteins for a 3 month (HDL) period and resultedlower intriglycerides a significant levels decrease [17]. ofMoreover, fasting plasma administration glucose, of cholesterol, 15 mg of saffron Low Density capsules Lipoproteins to diabetic patients (LDL-c), andfor LDLa 3 month/HDL ratioperiod in resulted comparison in a tosignificant the placebo decrease [18]. of fasting plasma glucose, cholesterol, Low DensityTo date, Lipoproteins well over (LDL-c), 6500 and publications LDL/HDL listed ratio in in comparison the PubMed to the Central placebo database [18]. demonstrate the potentialTo date, useswell over of sa 6500ffron publications extracts as listed adjuvant in the in PubMed anti-cancer, Central anti-diabetic, database demonstrate anti-depressant, the andpotential anti-inflammatory uses of saffron treatment. extracts as Ouradjuvant group in hasanti-cancer, shown theanti-diabetic, presence anti-depressant, of substantial amounts and anti- of kaempferolinflammatory
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