RevistaBrasileiradeFarmacognosia25(2015)553–566
w w w . s b f g n o s i a . o r g . b r / r e v i s t a
Review
Article
Carthami
flos
:
a
review
of
its
ethnopharmacology,
pharmacology
and
clinical
applications
Yanhua
Tu
a,1,
Yingru
Xue
a,1,
Dandan
Guo
a,
Lianna
Sun
b,∗,
Meili
Guo
a,∗aDepartmentofPharmacognosy,SchoolofPharmacy,SecondMilitaryMedicalUniversity,Shanghai,PRChina
bDepartmentofChineseMedicineIdentification,SchoolofPharmacy,SecondMilitaryMedicalUniversity,Shanghai,PRChina
a
r
t
i
c
l
e
i
n
f
o
Articlehistory:
Received1May2015 Accepted3June2015 Availableonline27June2015
Keywords:
Carthamiflos
Pharmacology Clinicalapplications Sideeffects
a
b
s
t
r
a
c
t
Carthamiflos,thedriedfloretofCarthamustinctoriusL.,Asteraceae(safflower),hasbeenwidelyusedin traditionalChinesemedicinetotreatabroadrangeofailments,suchascoronaryheartdisease,angina pectoris,gynecologicdisease,stroke,andhypertension.However,althoughseveralstudiesonCarthami floshavebeendoneconsecutively,theresultsareusuallyscatteredacrossvariousdocuments.Thisreview aimstoprovideup-to-dateinformationonthetraditionaluses,pharmacology,clinicalapplications,and toxicologyofCarthamiflosinChinaandtherebytoprovideabasisforfurtherinvestigationofitsuse totreatdissimilardiseases.VariousethnomedicalusesofCarthamifloshavebeendocumentedinmany ancientChinesebooks.CrudeextractsandisolatedcompoundsfromCarthamiflosshowabroadrange ofpharmacologicalproperties,suchasprotectiveeffectsonbraintissue,onosteoblasts,andin myocar-dialischemia,aswellasanti-inflammatory,antithrombotic,antitumor,andantidiabeticactivities.To date,safflowerandsaffloryellowinjectionshavebeenusedtotreatcoronaryheartdisease,chronic pul-monaryheartdisease,cerebrovasculardiseases,orthopedicdiseases,anddiabetesmellitus.Regarding thetoxicologyofCarthamiflos,amongthesideeffectsthathavebeenobservedareallergicreaction, spermatogeneticfailure,fattyliver,andnephrotoxicity.
©2015SociedadeBrasileiradeFarmacognosia.PublishedbyElsevierEditoraLtda.Allrightsreserved.
Introduction
Carthamiflos,thedriedfloretofCarthamustinctoriusL., Aster-aceae(safflower),hasbeencommonlyusedintraditionalChinese medicinetotreatawiderangeofailments,suchascoronaryheart disease,angina pectoris,gynecologicdisease,stroke,and hyper-tension.InChina,Carthamifloshasbeenusedasmedicationfor morethan2100years,sincethespecieswasintroducedbyZhang Qianinthehistoric“SilkRoad.”TraditionalChinesemedicinehas accumulatedponderableinformationontheuseofCarthamiflos,
whichhasbeendocumentedinancientmanuscriptsand
summa-rizedinrecentlypublishedbooks,suchasChinesePharmacopoeias, “NewlyRevisedCanonofMateriaMedica,”and“MedicalTreasures
oftheGoldenChamber.”Modernpharmacologicalinvestigations
havesubstantiatedthatCarthamiflosextractsorisolatedpure com-ponentshaveprotectiveeffectsonbraintissue,myocardialtissue, andosteoblasts,aswellasantithrombotic,anti-inflammatory,and antitumoreffects,which,tosomedegree,aretightlylinkedtothe
∗ Correspondingauthor.
E-mails:[email protected](L.Sun),[email protected](M.Guo).
1Theseauthorscontributedequallytothiswork.
accountsofbloodcirculationpromotion,bloodstasisremoval,and painreliefrecordedinancientChinesedocuments.
Thusfar,manycompoundshavebeenisolatedfromCarthami
flos,includingquinochalones,flavonoids,alkaloids,polyacetylene, aromaticglucosides,organicacids,etc.(GuoandZhang,1998,2000; Zhangetal.,2004,2005,2006,2009a;AsgarpanahandKazemivash, 2013; Zhou et al., 2014). Effective compounds or active parts ofCarthamifloshavebeenscreenedforpharmacologicalactivity
invivoandinvitro,suggestinggoodrepercussionsforhumanhealth
maintenanceandpromotion.Somecompoundshavebeenapplied
toclinicaltreatmentofcoronaryheartdisease,chronicpulmonary heartdisease,cerebrovasculardiseases,orthopedicdiseases,and diabetesmellitus.Thispaperintroducestheethnopharmacological application,pharmacologicalproperties,modernclinical applica-tion,andsideeffectsofCarthamiflosinChinaandtherebyprovides supportandevidenceforfurtherinvestigationofitsuse.
Ethnopharmacologicaluse
IntraditionalChinesemedicine,theapplicationofCarthamiflos
inclinicaltreatmentcanbetracedbacktoasearlyas2000years
ago. Thebook Medical Treasures oftheGolden Chamber,
writ-tenbyZhangZhongjin,documented that,whengiven asacold
http://dx.doi.org/10.1016/j.bjp.2015.06.001
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infusion,thedecoctionofCarthamiflosanddistillatespirit(1:10) hadtherapeuticeffectsongynecologicaldiseases,including
induc-tion of abortion early in pregnancy, expulsion of a retained
afterbirthorstillbirth,andmoderationofpainduringmenstrual periods.Givenasamajormedicamentportion,Carthamifloshas beenusedasaremedyforcoronaryheartdisease,anginapectoris, hypertension,andgynecologicaldiseases(GuoandZhang,1996, 1999).Theleavesareusedasadiuretic,anappetizer,andacurefor urinarydischarge.Also,powdermadefromleavesofsafflower com-biningwithCortexLyciiRadiciscouldbeusedtoclavusafteradding sesameoiltobepasty.Duetotheimpactofspecificgeographic environments,customs,andcultures,themedicinalpropertiesand clinicalapplicationsof Carthamiflosdifferacrossvariousethnic regionsofChina.InMongolianandTibetanmedicine,Carthamiflos
isusedtotreatlivermetabolismdisorders,suchashepatomegaly, hepaticinjury,xanthoeyes,andheathepaticblood(Luo,1988).In DaiandYimedicine,thenatureofCarthamiflosissimilartothat inChinesemedicine,butitsclinicalapplicationsaremainlyaimed attreatinginfertility,galacturia,softtissueinjuries,andfractures (Linetal.,2003;Guan,1993).Modernmedicalinvestigationshave showntheactivitiesofCarthamiflosinimprovingoxygensupply totheheartandbrain,mitigatingischemicinjury,and hepatopro-tection,which,tosomeextent,providescientificsupportforthe traditionalmedicinaltheoryandapplicationofCarthamiflos(Chen etal.,2012).
Pharmacologicalreports
Effectonbraininjury
HydroxysaffloryellowA(HSYA)(1), amajoractive chemical ingredientofCarthamiflos,hasbeenwidelyresearchedinChina asatreatmentforcerebrovasculardiseases(Lietal.,2005,2010; Zhangetal.,2009a,b;Fengetal.,2010;Tangetal.,2010). Numer-ousstudieshaveindicatedaprotectiveabilityofHSYA(1)against brainimpairment.Aninvitrostudycorroboratedthatinterference withHSYA(1)(0.072mg/ml)contributedtonerveregenerationof anorganotypic hippocampalslice fromneonatalSDratsin nor-malorischemicconditions(Qinetal.,2012a;Chart1).HSYA(1) alsorelievedoxygen-glucosedeprivationinneuralstemcellinjury andcontributedtoneurogenesisinvitro(Qinetal.,2012b;Chart 1).Neurondamageinducedbyexposuretoglutamateandsodium cyanide (NaCN) in cultured fetal corticalcells was significantly inhibitedbyHSYA(1)(Zhuetal.,2003;Chart1).Inaninvivostudy, thetherapeuticeffectofHSYA(1)onfocalcerebralischemiawas investigatedinamiddlecerebralarteryocclusion(MCAO)model. Beginningwitha doseof3mg/kg, HSYA(1)suppressed throm-bosisformationinMCAOrats,followedbyinhibitionofplatelet aggregationandadjustmentofPGI2/TXA2(Zhuetal.,2003,2005; Chart1).Spinalcordischemia–reperfusion injuryinrabbitswas alsofoundtoimprovewhentreatedwithHSYA(10mg/kg)(Shan etal.,2010;Chart1).Aninvestigationofitseffecton mitochon-drialpermeabilitytransitionpores(mtPTP)inratbrainindicated thatHSYA(1)(10–80mol/l)inhibitedCa2+-inducedswellingof
mitochondriaisolatedfromratbrainsandgenerationofROS.Taken
togetherwith the improvedmitochondrial energy metabolism,
theenhancedATPlevels,andtherespiratorycontrolratio,itwas extrapolatedthatHSYA(1)inhibitedtheopeningofmtPTPbyafree radical-scavengingactionin thebrain,which consequentlymay haveresultedintheneuroprotectiveeffect(Tianetal.,2008).In astudybyPanetal.(2012),HSYA(1)(5mg/kg,i.p.)wasfoundto attenuatebraininjuryinducedbylymphostaticencephalopathy, which showing alleviating neurologic deficits and cell apopto-sisintherostralventrolateral medulla(RVLM),suppressingthe
impaired regulatoryroles of theautonomic nervous system in
cardiovascular,andpreventingthedecreaseofendothelialnitric
oxide synthase (eNOS) mRNA. Additionally, pretreatment with
HSYA(1)improvedspatialmemorydeficitsandinhibitedchanges intheblood–brainbarrier,theSODactivity,andthe malondialde-hyde contentin brain injury induced by12C6+particle therapy
(Gan et al., 2012).A recent workimplied that HSYA (1)has a protective effect against cerebral I/Rinjury, partly by reducing
apoptosisthroughthePI3K/Akt/GSK3b signalingpathway(Chen
etal.,2013a,b;Chart1)anddecreasingnitrotyrosineformation(Sun etal.,2013).Basedonthesestudies,itwashypothesizedthatthe underlyingmechanismsoftheprotectiveeffectsofHSYAonbrain injuryinvolvedareductionoflipidperoxidation,thesuppressionof superoxidedismutase(SOD)andglutathioneperoxidase(GSH-Px) activities,theupregulationoftheexpressionofendothelialnitric oxidesynthase(eNOS) protein,anda decreaseincellapoptosis andstructuraldamageofnervoustissues.Recentreportshave sug-gestedthatHSYA(1)protectcorticalneuronsfrominhibitingthe phosphorylationofPPARrandtheexpressionofNR2B-containing NMDAreceptorsandregulatingtheBcl-2family(Yangetal.,2010; Liuetal.,2012a,b).However,whetherneuronalNOSisinvolved inthe protectiveeffects of HSYA(1)remains tobedetermined in a future study. Anin vitroinvestigation alsoconfirmed that HSYAshowedprotectiveeffects onneurotoxicityinducedby -amyloidinPC12cells,asevidencedbyreversedchangestriggered by-amyloid,suchasadecreaseincellviability,glutathionelevel, mitochondrialmembranepotential,andtheratioofBcl-2/Bax pro-teinexpression,alongwithanincreaseinlactatedehydrogenase, DNAfragmentation,andthelevelsofmalondialdehydeand intra-cellularreactiveoxygenspecies.ThisfindingsuggestedthatHSYA (1)wasapromisingcandidatedrugforpreventionandtreatment ofAlzheimer’sdisease(Kongetal.,2013a,b).
Investigationsontheprotectiveeffectsofextractsorother com-poundsfromCarthamiflosonthenervoussystemhavealsobeen done.Inaninvitrostudy,Zhaoetal.(2009a)foundthatallsolvents extracted fromCarthami flos,which containextracts of chloro-form,ethyl acetate,and n-butyl alcohol (1,10, and 100g/ml,
respectively),markedlyenhancedbothdopamineuptakeby
Chi-nesehamsterovary(CHO)cellsstablyexpressingthedopamine
transporter(DAT)andnorepinephrineuptakebyCHOcells
express-ing the norepinephrine transporter (NET), and simultaneously
depressed serotonin uptake by CHO cells expressing the
sero-tonin transporter(SERT),indicating thatextractsfromCarthami floswouldimproveneuropsychologicdisordersthroughregulating themonoamine-transporteractivity.Further,aninvitro investi-gationshowedthatN1,N5-(Z)-N10-(E)-tri-p-coumaroylspermidine
(2)potentlyandselectivelyinhibitedserotoninuptakeinS6cells
or in synaptosomes witha reversiblecompetitive property for
5HTuptakeinhibition,mirroringitseffectofimproving neuropsy-chologicdisordersthroughregulatingserotoninergictransmission (Zhao et al., 2009b; Chart 1).Carthamin (10mg/kg) (3)
signif-icantly decreased the formation of malondialdehyde in mouse
cerebrum and of thiobarbituricacidreactivesubstances and 8-hydroxy-2′-deoxyguanosine (8-OHdG) in the cerebral cortex of
ratssubjectedtoan injectionofFeCl3 solutionintothesensory
motorcortexinhibitedglutamate-induced(Hiramatsuetal.,2009;
Chart 1).Nicotiflorin (kaempferol-3-O-rutinoside) (4), a natural flavonoidextractedfromCarthamiflos,showedneuroprotection in focal cerebralischemia invitro and in vivo,which mightbe attributed to the upregulation of endothelial nitric oxide syn-thase (eNOS) activity (Li et al., 2006a,b; Chart 1). A study by
Huang etal. (2007)showed thatkaempferol-3-O-rutinoside (4) (30,60, and120mg/kg)significantly attenuatedtheincrease of lacticacidandmalondialdehyde(MDA)contentsandthedecrease inLDH,Na+K+ATPase,Ca2+Mg2+ATPase,andsuperoxidedismutase
(SOD) activity in multi-infarct dementia model rats,
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Chart1
Pharmacologicalactivitiesofcompounds/extractfromCarthamiflos.
Biologicalactivities Extract/compound(effective dose)
Testmodel Remarks Reference
Protectiononbrain injury
HSYA(1)(0.036and 0.072mg/ml)
OrganotypichippocampalslicesfromneonatalSD rats(invitro)
IncreaseorganotypichippocampalcellsfromneonatalSDrats (DAPI+,BrdU+andNestin+)witharrangedincordsandmigrated tothecodexascomparedtocontrol(normalsaline)
Qinetal.(2012a)
HSYA(1)(0.036and 0.072mg/ml)
Oxygen-glucosedeprivationinhippocampalslice culturesfromrats(invitro)
Reducetheproliferationofneuralstemcellcomparedtonormal saline
Qinetal.(2012b)
HSYA(1)(10–80mol/l) Ca2+-inducedandH
2O2-inducedswellingof
mitochondriaisolatedfromratbrains(invitro)
Inhibitedswellingandreactiveoxygenspeciesofmitochondria, improvemitochondrialenergymetabolismandenhancedATP levelsandtherespiratorycontrolratiocomparedto50mol/l Ca2+-treatedgroup
Tianetal.(2008)
HSYA(1)(3.0and6.0mg/kg, sublingularveininjection)
MaleWistar–Kyotoratswithmiddlecerebral arteryocclusion(MCAO)(invivo)
Decreaseneurologicaldeficitscoresandreducinginfarctarea comparedtothesalinegroupanddosageof6.0mg/kgshowa similarpotencyasnimodipine(0.2mg/kg)
Zhuetal.(2003)
HSYA(1)(3.0and6.0mg/kg, sublingularveininjection)
MCAOrats(invivo) Produceadose-dependentreductionininfarctarea(60%and85% respectively),increasetheratioof6-Keto-PGF1aandTXB2,reduce thromboticweight
Zhuetal.(2005)
HSYA(1)(10.0mg/kg,ear intravenousinjection)
Rabbitswithischemia/reperfusion(I/R)injury (invivo)
AttenuateI/Rinducednecrosisinspinalcords,alleviateoxidative stressasindicatedbydecreasedmalondialdehyde(MDA)leveland increasedsuperoxidedismutase(SOD)activityandprotected neuronsfromI/R-inducedapoptosisinrabbits
Shanetal.(2010)
HSYA(1)(5mg/kg,i.p.) Ratswithlymphostaticencephalopathy-induced braininjury(invivo)
Alleviatetheneurologicaldeficits,attenuatecellapoptosisinthe rostralventrolateralmedulluscomparedtosalinegroup
Panetal.(2012)
HSYA(1)(5,10and20mg/kg, intraperitonealinjection)
Mousewithbraininjuryinducedby12C6+particle
therapy(invivo)
Dose-dependentlyimprovethespatiomemorydeficitsand increaseSODactivityandreducemalondialdehydecontentin braintissue
Ganetal.(2012)
HSYA(1)(4and8mg/kg, tail-veininjection)
MCAOrats(invivo) DiminishthenumberofapoptoticcellsandincreasetheBcl-2/Bax ratioaswellasthephosphorylationsofAktandGSK3b
Chenetal.(2013a,b)
N1,N5-(Z)-N10-(E)-tri-p
-coumaroylspermidine(2) (0.04–100M)
Chinesehamsterovarycells(invitro) InhibitserotoninuptakeinS6cells(IC50=0.74±0.15M)andin synaptosomes(IC50=1.07±0.23M)
Zhaoetal.(2009b)
Carthamin(3)(10mg/kg) Ratswithepilepticfociinducedbyiron(invivo) Inhibit8-hydroxy-2′-deoxyguanosineinthecerebralcortexofrats Hiramatsuetal.(2009)
Kempferol-3-O-rutinoside(4) (2.5,5and10mg/kg,tailvein)
Ratswithpermanentfocalcerebralischemia (invivo)
Dose-dependentlyreducebraininfarctvolumeandneurological deficitscomparedwithnimodipin(positivecontrol)
Lietal.(2006a)
Kempferol-3-O-rutinoside(4) (25–100g/ml)
Culturedneuronssufferedhypoxia(invitro) Attenuatecelldeathandreducelactatedehydrogenaserelease Lietal.(2006a)
Anti-myocardial ischemiaeffect
Ethanolicextract(62.5and 125g/ml)
H9c2cardiomyoblastcells(invitro) ReduceIkBdegradationandNFkBactivation,activateof anti-apoptoticproteins,Bcl-2andBcl-xL,stabilizationthe mitochondriamembraneandthedown-regulationofextrinsicand intrinsicpro-apoptoticproteins,suchasTNF␣,activecaspases-8,9 and3t-Bid,Bax,comparedtoLPS
Tienetal.(2010)
HSYA(1)(4and8mg/kg,tail vein)
Ratswithcoronaryarteryligation(invivo) Reductionofmyocardialinfarctionsize,superoxidedismutase activity,endothelialnitricoxidesynthaseactivityandnitricoxide content,andinhibitelevationofcreatinekinaseactivityand malondialdehydecontent
Wangetal.(2009a,b)
HSYA(1)(0.1–3mg/kg, intravenousinjection)
Ratswithpentobarbitone-anesthetized normotensiveandspontaneouslyhypertensive (invivo)
Dose-dependentlyreduceheartrate,meanarterialpressure,left ventricularsystolicpressure,leftventricularend-diastolicpressure
Nieetal.(2012)
N-(p-coumaroyl)serotonin(6) (5×10−7M)
Modelofperfusedguinea-pigLangendorffhearts subjectedtoischemiaandreperfusion(invitro)
IncreasetheNOlevelattheendofischemiaandshow63.2% recoveryrateofleftventriculardevelopedpressurecomparedto drug-freecontrol(30.8%)
Hottaetal.(2002)
N-feruroylserotonin (5×10−7M)(7)
SimilartoN-(p-coumaroyl)serotonin Show61.0%recoveryrateofleftventriculardevelopedpressure andquenchtheactivityofactiveradicals
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Chart1(Continued)
Biologicalactivities Extract/compound(effective dose)
Testmodel Remarks Reference
Antithrombotic effect
Aqueousextract(1and 0.7g/kg,oral)
(a)Ratsarterialthrombosismodel(invivo) (b)Ratsvenous(Wessler)thrombosismodel (invivo)
(c)Mouseswithcollagen/epinephrine-induced pulmonaryembolism(invivo)
(d)Mousestailbleeding(invivo)
(a)Showamildthrombosisinhibitionbutwithoutdifferencewith controlwhenplusclopidogrelornot
(b)Inhibitthrombusformationfrom16.1to7.9mgandledtoa significantdecreaseinvenousthrombusweightwhenadded clopidogrelandalsoaugmentthrombintimeandprothrombintime (c)Increasethenumberofnon-paralyzedanimalsto33.3%and survivingratesto53.3%
(d)Prolongedthebleedingtimewhenaddedclopidogrel
LiandWang(2010)
Carthaminsyellow(100and 200mg/kg,oral)
Ratswithbloodstasis(invivo) Decreasethewholebloodviscosity,plasmaviscosity,erythrocyte aggregationindex,hematocritandplateletaggregationwhen comparedwithaspirin
Lietal.(2009)
HSYA(1)(1×10−4M) HumanECline(EAhy926)(invitro) Attenuatecellcyclearrestandinhibitcellapoptosisina
concentration-dependentmannerandincreasethebcl-2/baxratio, VEGFproteinconcentration,VEGFmRNAexpression,HIF-1aprotein accumulationanditstranscriptionalactivity
Jietal.(2008)
HSYA(1)(1×10−6,1 ×10−5
and1×10−4M)
Humanumbilicalveinendothelialcells(HUVEC)
(invitro)
InhibitcellapoptosisandcellcycleG1arrestinducedbyhypoxia IncreasetheBcl-2/BaxratioofproteinandNOcontentofcell supernatant
Reducep53proteinexpressionincellnucleus
Jietal.(2009)
Anti-inflammatory Saffloryellow(25and 50mg/kg,intraperitoneal)
Ratsofpulmonaryfibrosisinducedbybleomycin (invivo)
(a)Alleviatethelossinbodyweight,theincreaseofhydroxyproline contentinthelungtissuesandpathologicchangesofpulmonary fibrosis
(b)Preventtheincreaseofa␣-SMApositivecellsandTGF-bb1 expression
Wangetal.(2011a,b)
Saffloryellow(0.05,0.25and 1.25mg/ml)
Humanembryolungfibroblast(invitro) Dose-dependentlyinhibittheelevationof␣-SMAexpressionandthe morphologicalchange
Wangetal.(2011a,b)
Methanolextract(10,50and 100g/ml)
RAW264.7macrophages(invitro) TheincreaseHO-1proteinexpression,thereducedproductionof NO,PGE2,iNOSandCOX-2,andtheinhibitionof(TNF)-␣-mediated
VCAM-1expressionandNF-Bluciferaseactivity
Junetal.(2011)
HSYA(1)(1,4and16mol/l) HumanalveolarepithelialA549cells(invitro) (a)InhibittheexpressionofTLR-4,Myd88ICAM-1,TNF␣,IL-1and IL-6
(b)Inhibitthephosphorylationofp38MAPK,theadhesionof leukocytestoA549cellsanddecreaseNF-Bp65nuclear translocation
Songetal.(2013)
HSYA(1)(5×10−6,10 ×10−6
and20×10−6mol/l)
UmbilicalveinendothelialEahy926cells(invitro) InhibittheincreasedexpressionofTLR-4,IL-6,IL-1andTNF␣ Zhuetal.(2012)
HSYA(6,15and37.5mg/kg, i.v.)
MicewithLPS-inducedpulmonaryinflammatory injury(invivo)
(a)Amelioratepulmonaryedemaandinflammatorycellinfiltration (b)Suppressp38MAPK,NF-Bp65activationandalter
inflammatorycytokineexpression(TNF-␣,IL-1,IL-6andIL-10)
Sunetal.(2010)
HSYA(1)(26.7,40and 60mg/kg/day,intraperitoneal injection)
Micewithbleomycin-inducedpulmonaryinjury (invivo)
Attenuatethelossinbodyweight,theincreaseofmyeloperoxidase activityandpathologicchangesofpulmonaryinflammation (c)AttenuatetheincreasedexpressionofTNF-␣,IL-1andTGF-1, theincreasedactivationofNF-Bandphosphorylationofp38MAPK
Wuetal.(2012a,b)
Hepatoprotective effect
HSYA(1)(5mg/kg/dayfor12 weeks,oral)
RatswithCCl4-inducedhepaticfibrosis(invivo) Thedecreaseinfibrosisandexpressionof␣-SMAprotein,MEF-2C
gene,T-RI,T-RII,MEKK3,MEK5,andphosphorylationofERk5
Zhangetal.(2011,2012a,b,c)
HSYA(1)(10mg/kg, intraperitonealinjection)
Ratswithhepaticfibrosisinducedbyoxidative stress(invivo)
(a)Increasetheactivitiesofantioxidantenzymesandreduce␣-SMA level.
(b)Up-regulatingtheexpressionofPPARrandMMP-2,anddown regulatingtheexpressionofTGF-1andTIMP-1
Wanetal.(2013)
Carthamusred(5,10and 20mg/kg,oral)
RatswithCCl4-inducedhepaticfibrosis(invivo) (a)LowertheserumlevelsofALT,AST,ALPandtotalproteininliver
damageratmodels
(b)Up-regulateNrf2,GST␣andNQO1expressionswereatthe proteinlevel
(c)ElevatetheactivitiesofantioxidantenzymesandlevelofGSH andlessenthecontentofTBARScomparedtosilymarin
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energymetabolismfailure,andoxidativestressthatareinvolved in multi-infarctdementia. Additionally, postischemic treatment withkaempferol-3-O-glucoside(7.5mg/kg)(5)showeda neuro-protectiveeffectinfocalcerebralischemia–reperfusionrats.The mechanismofthesekaempferolflavonoideffectswasattributed to anti-neuroinflammatory activity by inhibiting the activation
ofSTAT3andNF-kBp65,includingindependentand dependent
pathwaysof IkBdegradation and thesubsequent expressionof
pro-inflammatorymediators(Yuetal.,2013).Nevertheless,more pharmacologicalevaluationsinanimalmodelsrelatingto neuro-logicaldisordersneedtobeconsideredinfuturestudiestoexplain
theunderlyingmechanismsofthesecompounds.
Effectsonmyocardialischemia
EthanolicextractofCarthamiflos(62.5g/ml)hastheabilityto suppressJNKactivityandinhibitLPS-inducedTNF␣activationand apoptosisinH9c2cardiomyoblastcells(Tienetal.,2010;Chart1). Inaninvivostudy,theprotectiveeffectsofapurifiedextractfrom
Carthamiflos(100,200,400,and600mg/kgbodywt.)on
myocar-dial ischemia wasassessed in a model of myocardial ischemia
injuryinducedbyleftanteriordescendingcoronaryartery(LAD) occlusion,which resulted inreduced infarctsize and improved cardiac function (Han et al., 2009).Further investigations have reportedthatthiscardioprotectiveeffectofCarthamiflosextract (200mg/kg)wasnot onlysupportedbydecreased levelsof cre-atinekinase(CK)andLDHbut,further,couldbestrengthenedby addingPanaxnotoginseng(Burk)F.H.Chen(EPN)extract(50mg/kg) (Han et al., 2013a,b). HSYA (1) (4 or 8mg/kg) also showed a cardioprotectiveeffect,asevidencedbythereducedmyocardial infarctionsizeinratswithacutemyocardialischemiainducedby LADligation(Wangetal.,2009a,b).Whethertheprotectiveeffect ofHSYA(1)onmyocardialischemiainjurycouldbeenhancedby
combination with EPN remains inconclusive (Fu et al., 2011). Contrarily, extracellular Ca2+ influx through receptor-operated
Ca2+channels and potential-dependent Ca2+channels could be
blockedbycrudedrugofCarthamiflos(Liuetal.,2005).Afurther findingshowedthatHSYA(1)markedlyreducedCa2+influxon
car-diaccells,aswellasdecreasedthecontractileforceandheartrate inrats.SuchaneffectmayimplicatetheactivationofBKCaandKATP
channels(Nieetal.,2012;Chart1).WhetherHSYA(1)couldlower theperipheralresistanceremainsunderstudy.Othercompounds, such asN-(p-coumaroyl) serotonin(6) and N-feruroylserotonin
(7), showed cardioprotective effects on isolated guinea pig
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Antithromboticeffect
TheaqueousextractsofCarthamifloshavebeenshowntobe moreefficientthanclopidogrelagainstvenousthrombosisand pul-monaryembolism,withsimilarfunctionstocarthamusyellow(Li andWang,2010;Lietal.,2009;Chart1).AnHSYA(1)activityof enhancingthesurvivalofvascularendothelialcellsunderhypoxia hasalsobeenfound,whichmaybecorrelatedwithitseffecton upregulationoftheHIF-1a-VEGFpathwayandregulationof Bcl-2/Bax (Jiet al., 2008; Chart 1).A further investigationshowed thatHSYA(1)couldprotecthumanumbilicalveinendothelialcells (HUVECs)fromhypoxia-inducedinjurybyinhibitingcellapoptosis andcellcyclearrest,partlyindicatingthemolecularmechanism ofHSYA(1)inthetreatmentofischemicheartdisease(Jietal., 2009; Chart 1).Contrarily,carthamin (3)(10mg/l)could repair theblockedHUVECmigrationandrefinementofthef-actin struc-turecausedbymodeledmicrogravity(MMG),whichwouldprovide anewalternativeforinterventionincardiovasculardysfunction apartfromHSYA(1).
Anti-inflammatoryeffect
InastudybyWangetal.(2010),Carthamiflosaqueousextract andcarthamusyellow(CY)wereexaminedregardingtheireffects
onLPS-inducedinflammatoryresponseinamurinemacrophage
celllineRAW264.7model.Accordingtotheresults,aqueousextract fromCarthamiflos(1–1000g/ml) andCY (1–2000g/ml)
sup-pressed the production of NO, PGE2, and IL-1 and decreased
theiNOSandcyclooxygenase-2(COX-2)proteinexpressionlevels
inLPS-inducedRAW264.7macrophages.Basedontheinhibition
ofcytosolIB-␣proteindegradationandphospho-NF-Bprotein expression,itwastheorizedthataqueousextractfromCarthami flosandCY mayinhibitLPS-stimulatedexpressionsof theiNOS andCOX-2genesthroughtheinactivationofNF-B.Asan effec-tivepartoftheaqueousextractofCarthamiflos,saffloryellow(SY) hasshownaninhibitoryeffectonpulmonaryfibrosisinvivoand
invitro,supportedbysuppressingtheexpressionofa-smooth mus-cleactin(a-SMA)(Wangetal.,2011a,b;Chart1).Regardingthe anti-inflammatoryactionofmethanolextractsofsafflower(MEC), ithasbeenreportedthatMECtriggeredhemeoxygenase-1 expres-sionthroughNrf2(NF-E2-relatedfactor)translocationandNF-kB activityinhibition.Thispotentialmolecularmechanismhas
pro-videdother clues on themolecular mechanism underlyingthe
anti-inflammatoryaction of aqueousextract from Carthamiflos
andCY(Junetal.,2011;Chart1).Contrarily,polysaccharidesfrom
Carthamifloswerefoundtohaveimmunomodulatingactivitiesthat effectivelyactivatedtheNF-kBsignalingpathwaythroughTLR4and inducedtheproductionofvariouscytokines(IL-1,IL-6,IL-12,and IFN-␥)byperitonealmacrophages(Andoetal.,2002).
RecentinvestigationsonHSYA(1)havefocusedonthe treat-mentof acutelung injury (ALI).The effectsof HSYA(1)onALI havebeenevaluatedinvitro(humanalveolarepithelialA549cells andumbilicalveinendothelialcells(Eahy926cells))andinvivo
(LPS-inducedand BLM-induced ALImice), which allmanifested
thatHSYA(1)amelioratedacutelunginjurybysuppressingboth p38MAPK(mitogen-activatedproteinkinase)phosphorylationand NF-Bactivation,subsequently leading toa dramatic reduction ininflammatory cellinfiltration andpro-inflammatory cytokine expressioninlungtissue,aswellaspulmonaryedemaand respi-ratorydysfunction(Sunetal.,2010;Songetal.,2013;Zhuetal., 2012;Wuetal.,2012a,b;Chart1).Duetothehighwatersolubility ofHSYA(1),thesefindingsimplythatHSYA(1)maytargetthecell membraneandtheninterferewiththeinterplayofreceptorsand theirspecificligands(suchasmicrobialligands,pro-inflammatory cytokines, growth factors, etc.) to regulate downstream signal transduction pathwaysso as to exert its effects. However, the
concretemechanismbywhichHSYAaltersintracellularsignaling stillwarrantsfurtherstudies.
Othercompoundshaveanti-inflammatoryeffectsaswell. Com-paredwithginkgolideB(IC505.45×10−6mol/l),saffloquinosideA
(8)(10−5mol/l)exhibited54.3%inhibitoryrateonthereleaseof -glucuronidasefromratpolymorphonuclearneutrophils(PMN), whichwasinducedbytheplatelet-activatingfactor(PAF), suggest-ingitsanti-inflammatoryactivity(Jiangetal.,2010).
Antitumoreffect
Carthamifloshasbeenappliedin canceradjuvanttherapy in
traditionalmedicine.Modernpharmacologicalexperimentshave
confirmedtheantitumoractivityofCarthamiflosinvitroandin vivo.HerbalextractofCarthamiflos(40mg/ml)has antiprolifera-tiveandproapoptoticeffectsonhepaticstellatecells,whichmay actbyregulatingthegeneexpressionofFasandBcl2 pathways (Choretal.,2005).Furtherinvitroexperimentshaveshownthat saffloryellowB(9)(1,10,and 100nmol/l)protected pheochro-mocytoma(PC12)cellsfromH2O2-inducedinjuryandapoptosis
throughantioxidantandantiapoptoticmechanismsthatarelinked tosuppressingcaspase-3activityandBaxexpressionand increas-ingBcl-2synthesis(Wangetal.,2009a,b).Anotherinvitrostudy by Zhang et al. (2012a,b,c) reported that polysaccharide S of
Carthami flos (0, 0.02, 0.04, 0.08, 0.16, 0.32, 0.64, and 1.28g/l)
couldrestraintheproliferationofSMMC-7721andenhancethe
apoptosisof SMMC-7721 in a dose- and time-dependent
man-ner,asshownbytheincreasedBaxexpressionandthedecreased
Bcl-2 expression and mitochondrial membrane potential.
How-ever,HSYA(1)(0.028g/l)inhibitedthegrowthofatransplanted BGC-823 tumorthrough inhibitionof tumorvascularization (Xi et al., 2012). Carthamin (3)dose-dependently (10−5, 10−4, and
10−3mol/l)inducedtheK562leukemiccellstothehaemoglobin
endcells, themechanismbehindwhich remainsunknown(Wu
etal.,2012a,b).Invivo,Changetal.(2011)showedinvivothe anti-tumoractivitiesofCarthamiflos-treateddendriticcellvaccinein
JC(mousemammaryadenocarcinoma)tumor-bearingmice,
rele-vanttothepolarizationtowardTh1cytokinesandtheincreasein cytotoxicTlymphocytes.Thisfindingfurthersupportsthereport regardingtheeffectivenessofCarthamiflosinbreastcancer(Loo etal.,2004).Asdeterminedfromthesestudies,themechanism responsiblefortheantitumoractivityofCarthamiflosmayoccur partlybysuppressingcaspase-3activityandBaxexpressionand byincreasingBcl-2synthesis,aswellasbyinhibitingtumor vas-cularization. Inhibition of human tyrosinase activity increased withincreasingconcentrations ofkinobeonA (10)withtheuse of l-tyrosine or l-3,4-dihydroxyphenylalanine (l-DOPA) as the
substrate(Kanehiraetal.,2003a,b).Recentworkhasuncovered thatcarthamusyellowreducedtheactivityofmushroom
tyrosi-nase in a dose-dependent manner(IC50 1.01±0.03mg/ml) and
showedamodeofcompetitiveinhibitionwithaKiof0.607mg/ml.
Y.Tuetal./RevistaBrasileiradeFarmacognosia25(2015)553–566 559
production of B16F10 melanoma cells at concentrations of
4.0mg/ml, indicating no cytotoxicity (Chen et al., 2013a,b).As derived from these studies, Carthami flos has the potential to becomeausefulskin-whiteningagentorapotentnatural tyrosi-naseinhibitorinthefuture.
Effectonosteoblasts
In a study by Choi et al. (2010), extract of Carthami flos
(2–10g/ml)wasshowntoinhibitosteoclastogenesisby modu-latingthereceptoractivatorofnuclearfactor-kBligand(RANKL) signalinginMC3T3-E1cells.Recently,evaluationsoftheeffectsof othercompoundsonbonehavebeencarriedout.Liuetal.(2011a,b)
foundthat SY (1.6mg/ml) promoted therepair of injured
ten-doninLeghornchicken,manifestedbytheenhancedexpression
of bFGFand collagentype Iprotein.Also, HSYA (1) downregu-latedtheexpressionofTLR4mRNAcallusosteoblasts(LuandTu, 2012).Theseresultsmayhavetherapeuticimplicationsintreating osteoporosisandotherboneerosivediseases,suchasrheumatoid arthritisormetastasisassociatedwithboneloss.
Othereffects
HSYA(1)hasbeenfoundtoalleviatecarbontetrachloride(CCl4
)-inducedliverfibrosisinrats,inpartthroughinhibitionofhepatic stellatecell(HSC)activationandMAPkinaseextracellular regu-latedkinase5(Erk5)signaling(Zhangetal.,2011,2012a,b,c;Chart 1),suggestingthatHSYA(1)cantargetfibrogenicpathwaysand thereforemaybea potentialtherapy forhepaticfibrosis.A fur-therworkhasreflectedthattheprotectiveeffectofHSYA(1)on hepaticfibrosisinducedbyoxidativestressrequirestheactivation ofPPARr(Wanetal.,2013;Chart1).Regardingothercompounds, carthamusredhasbeenreportedtohaveahepatoprotectiveeffect inratswithCCl4-inducedliverdamage,whichmightbemediated
byinductionofantioxidantdefensethroughincreasedactivation oftheNrf2pathway(Wuetal.,2013;Chart1).
Compared with glibenclamide, hydroalcoholic extract
of Carthami flos (200mg/kg) has an antidiabetic activity in
alloxan-induceddiabeticrats,asshownbytheirdecreasedfasting bloodsugar,triacylglyceride,cholesterol,LDL-C,andVLDL-Clevels, aswellincreasedinsulinlevels(Asgaryetal.,2012).Diabetesis
usually accompanied by increased production of free radicals
or impaired antioxidant defenses (Cuerda et al., 2011; Kuroki etal.,2003;Maritimetal.,2003).Previousstudieshavedisclosed
Carthamiflosconstitutedagoodsourceofantioxidantcompounds with free radical-scavenging potential (Choi et al., 2010; Zhao et al., 2012; Hiramatsu et al., 2009; Kanehira et al., 2003a,b; Kambayashi et al., 2005), suggesting that it could beuseful in thepreventionofdiseasesinwhichfreeradicalsareimplicated.
Considering their effects on these lipid components and their
antioxidantactivity,Carthamiflosand itsactivecompoundscan
be assumed to be potential hypolipidemic agents that could
yield considerable advantages for both the diabetic condition
andtheassociatedatherosclerosisorhyperlipidemicconditions. Recently, an in vitro study further corroborated that HSYA (1)
(10–100mol/l) ameliorated methylglyoxal-induced injury in
culturedhumanbrainmicrovascularendothelialcells,asshown
bythedecreasedexpressionofcaspase-3and theaccumulation
ofadvancedglycationendproducts.Theseresultsarereminiscent ofthepotentialofHSYAasanovelstrategyforprotectingagainst vascularcomplicationsassociatedwithdiabetes(Lietal.,2013).
Liuetal.(2012a,b)reportedthatadministrationofCarthamiflos
toethyleneglycol(EG)-fedratsledtoasignificantreductionin CaOxcrystalformation,indicatingitsantilithiceffect.Meanwhile,
saffloweryellow(50mg/l)canprotecthumanrenaltubular epithe-lialcell lines (HK-2cells) fromdamage by inhibiting apoptosis inducedbyaristolochicacid,whichmaybeaffectedbysuppressing theactivationofcaspase-3.Thesefindingssuggestthatsafflower yellow may be beneficial to the treatment of aristolochic acid nephropathy.
Anin vivo studyby Luet al. (2008) showedthat
menopro-gen,aherbalformulaconsistingofLyciifructus,Rehmanniaeradix,
Morifructus,andCarthamiflos,significantlyincreasedthelevelsof serumestradiolandprogesteronebutreducedthelevelsof follicle-stimulatingandluteinizinghormonesinrats.
The5a-reductaseinhibitoryandhairgrowth-promoting activ-ities of Carthami flos ethanolic extract were tested in C57BL/6
mice, which resulted in a finasteride equivalent 5a-reductase
inhibitory activity (FEA)value of 24.30±1.64mg per 1g crude extract(Kumaraetal.,2012).Thisfindingmayleadtonew alterna-tivemedicinesforhairlosspreventionandtreatment.
Aninvitrostudyoftheanti-gammaherpesvirusactivityofn -hexaneandEtOHfractionsofCarthamiflosextracts(iSLK-BAC16 andiSLK-purocells)byLeeetal.(2013)indicatedthatn-hexane andEtOHfractionsofCarthamiflosextractscriticallyinfluenced twostagesoftheKaposi’ssarcomaherpesvirus(KHSV)lifecycleby abnormallyinducingKSHVlyticreactivationandseverely preven-tingKSHVvirionreleasefromtheviralhostcells.Simultaneously, themechanismofdysregulationofKSHVreplicationbyCarthami flosextractsmaybemediatedbydysregulatingthecellcycleand producing strong cytotoxicity.Based on this finding,studies to gathermoreinvitroandinvivoevidenceontheanti-gamma her-pesvirusactivitiesofCarthamiflosextractsandthecausesofcellular cytotoxicity are needed.In vivo, Carthami flosextract (200 and 400mg/kg)showedsimilarreductionsinthevolume,freeacidity, andtotal acidityofgastricsecretioninducedbycarbachol,such ascimetidineandverapamil,indicatingitsantiulcerogeniceffect (Mandadeetal.,2012).AsreportedbyLiuetal.(2005),Carthami floshasanaturalcalciumchannelblockingactivity,whichmayhave contributedtoitsantiulcerogeniceffect.Furtherstudiesto evalu-atetheexactmechanismofthiseffectaresuggested.Recently,the photoprotectiveactivityoftopicalHSYA(1)(100and200gper mouse)wasinvestigatedinaUV-inducedphotoagingmicemodel.
560 Y.Tuetal./RevistaBrasileiradeFarmacognosia25(2015)553–566
whichcouldpossiblybeattributedtotheantioxidativeproperty ofHSYA(1)andactivatedbypromotingendogenouscollagen syn-thesis(Kongetal.,2013a,b).
Clinicalapplications
Treatmentofcoronaryheartdisease
InastudybyHuang(2013;Chart2),patientswithcoronaryheart disease(CHD)treatedwithsafflowerinjectionshowedadecreasein hemodynamicparameters,containingthewholeblood,highshear viscosity,shearwholebloodviscosity,wholebloodviscosityand plasmaviscosity,whichwerelowerthanthoseofthecontrolgroup treatedwithSalviamiltiorrhiza injection,suggesting theclinical efficacyofsafflowerinjectionincoronaryheartdisease.Inanother study,safflowerinjectionwasfoundtoimprovetheclinical symp-tomsofanginapectorisandtheelectrocardiogramofCHDpatients
compared withgroups treated only withconventional western
medicine(SuandChai,2011;Chart2).Themajoractiveingredient ofCarthamiflos,saffloryellow,hasalsobeenusedtotreat coro-naryheartdisease.AstudybyLiuetal.(2011a,b)foundthatsafflor yellowcouldreducetheendothelin,matrixmetalloproteinase-9, andhigh-sensitivityC-reactiveproteinofdifferentpatientswith coronaryarterydisease(Chart2).Otherstudieshaveshownthat saffloryellowcoulddecreaseneuropeptideY,whichwas impor-tanttoimprovethepathogeneticconditionofpatients(Liuetal., 2008;Chart2).
Treatmentofchronicpulmonaryheartdisease
Theuseofsafflowerinjectiontotreatheartfailureinpatients withchronicpulmonaryheartdisease(CPHD)resultedinimproved heartfunction,asprovedbytheameliorativebloodgas parame-terscomparedwiththecontrolgroup(p<0.05)(Liao,2012;Chart 2).contrarily,recentstudiesonSYandSYinjectionhaveindicated theircurativefunctioninpulmonaryheartdisease.An investiga-tionbyChenetal.(2014)showedtheefficacyofSYinimproving STsegmentdepression(p<0.01)anddecreasingthelevelsofTnI, CK-MB,and CRP(p<0.05),therebyprotectingmyocardial injury fromCPHD(Chart2).Meanwhile,thepulmonaryfunction,aswell asbloodviscosityandrheology,ofpatientswithCPHDwasfound tobeamelioratedaftertreatmentwithSYinjection(RenandMei, 2014;Chart2).
Treatmentofcerebrovasculardiseases
Extractsandcompoundsfromsafflowerhavebeenappliedwith beneficialeffectsintheclinicaltreatmentofcerebrovascular dis-eases. Systemic evaluations of the clinical effect and safety of safflowerinjectioninthetreatmentofacuteischemicstrokehave indicatedthat suchinjectionis helpfulinimprovingneurologic functionaldeficitsandhasagoodsafetyprofile,asshownbythe meta-analysisresultsontotaleffectiverates,therelativerisk99% credibleregion(99%CI),thenumberoftreatmentsneeded(99%CI), andtheweightmeandeviation(99%CI),whichare1.19(1.10,1.28), 7.14(5.00,12.5),and−0.62(−1.10,−0.15),respectively(Maetal., 2012).Thelatentmechanismwaspartiallyinvolvedindecreasing theserumratioofIL-6/IL-10andsuppressingthelipidperoxidation byincreasingthelevelofsuperoxidedismutase,choramphenicol acetyltransferase,andmalondialdehyde,whichweresupportedby clinicalevidencefromLuoetal.(2014;Chart2).Also,injectionsof saffloweryellowandHSYAhavebeenreportedtobemore effec-tiveinclinicaltreatmentofacuteischemicstrokecomparedwith Ginkgoleafextractanddipyridamoleinjection(XiaoandHu,2011; Xingetal.,2008;Chart2).
Treatmentoforthopedicdiseases
Recently,clinicalobservationsofsafflowerinjectionin ortho-pedicdiseaseshavebeengraduallycarriedout,withgoodecho. Safflowerinjectionforpatientswithacutegoutyarthritisshowed equalefficacytotreatmentwithcolchicine,suggesting that saf-flowerinjectionmaybeanewalternativetreatmentforacutegouty arthritispatientsduetoitslowtoxicity(Lietal.,2011;Chart2).A clinicalreportbySuietal.(2011)pointedtotheapplicationof saf-flowerinjectioninisolatedlimbreplantation,whichresultedin95% survivalofreplantedfingerswithnoadverseeffect,exceedingthe 86%survivalrateinthecontrolgroup.Anobservationofits preven-tionofpostoperativetendonadhesioninflexortendoninjuryhas alsobeenconducted,whichresultedin60patientsbeinghealedin stageIandshowinganadvantageoverthedickonbiologicalfilm groupintermsoftotalactivemotionofthetendonafterfourweeks.
Noobviousdiscrepancyinpowertograspwasobserved,which
wasreminiscentoftheapotropaiceffectofsafflowerinjectionon postoperativetendonadhesioninflexortendoninjury(Choietal., 2013).
Treatmentofdiabetesmellitusanditscomplications
InaclinicalreportbyWei(2011;Chart2),safflowerinjection wasextrapolatedtoeffectivelypostponethedevelopmentofrenal failurecausedbydiabeticnephropathyonthebasisofroutine treat-ment.Similarly, theapplicationof saffloweryellowinjectionin diabeticnephropathyobtainedbeneficialresultsinpalliatingurine proteinandloweringtheserumcreatininelevelofpatients(Yang etal.,2011;Chart2;Qiuetal.,2013).Othercomplicationsof dia-betesmellitusthatcouldbetreatedwithsaffloweryellowinjection have beenreported.Diabeticperipheralneuropathy, a frequent chroniccomplicationofdiabetesmellitus,couldbeamelioratedby saffloweryellowinjection,whichmaybeimplicatedin ameliora-tionofmicrocirculationinpatients(YangandDai,2010;Chart2).In arecentclinicalobservation,saffloweryellowinjectionwasfound todecreasetheincidenceofdelayedgraftfunctionand thereby improverecoveryofgraftfunctionafterrenaltransplantation(Pang etal.,2014;Chart2).
Sideeffects
Aninvivostudyoftheacutetoxicityofcarthamusredshowed notoxicityandmortalityfordosesofupto2000mg/kg(Wuetal., 2013).However,somesideeffectshavebeenreportedinanimal
andhumanmodels.Afterbeinggivenorally(bygavagemethod)
tomiceatadoseof200mg/kgfor35consecutivedays,Carthami flosextractengenderedtheformationofmultinucleatedgiantcells inthegerminalepitheliumandresultedinamarkeddecreasein
epithelial vacuolization, germsloughing and detachment,
sem-iniferous tubule diameter, seminiferous epithelium height, and maturationarrest,suggestingitsabilitytochangethetestis histo-logicstructureandcausespermatogeneticfailure(Mirhoseinietal., 2012;Bahmanpouretal.,2012).Therefore,precautionsshouldbe takenwhenusingCarthamiflosextractonmenwhoareinfertile orhavereproductivedisorders.Testsofactivesystemic anaphy-laxisandpassivecutaneousanaphylaxisbyCarthamiflosinjection haveshownpositivereactionsinguineapigsandSDrats, respec-tively,indicatingitssensitizationtoallergicreaction(Zhangetal., 2012a,b,c).Anessentialrequirementforfuturestudiesisthe explo-ration of the recessive sensibilizing substance in Carthami flos
Y. Tu et al. / Revista Brasileira de Farmacognosia 25 (2015) 553–566 561 Chart2
Summaryofclinicaltrials.
Author(Date) Designduration Condition No.ofparticipants
randomized(age)
Intervention Comparison Mainresultsbetween
groups
Adverseevents Authorsconclusion
Huang(2013) 40days Coronaryheart disease(CHD)
122(57.33±15.02) Injectedwith20ml safflowerinjection with0.5%250ml glucoseinjectiononce aday
Salviainjection Thehemodynamic parameters,werelower thanSalviainjection (p<0.05)
Nonereported Safflowerinjection hasefficacyinCHD andcouldimprove heartfunction
SuandChai (2011)
2weeks CHD 100;50pergroup (42–75years)
Usualwestern medicinetreatment andIntravenouswith 40mlsafflower injectionwith0.5% 250mlglucose injection
Usualwestern medicinetreatment
Theclinicalsymptomsand electrocardiogramresults intreatedgroupwere betterthanthegrouponly treatedwithwestern medicine(p<0.05)
Nonereported Safflowerinjection couldmakeusual westernmedicine treatmentbetterin aspectof controllingangina pectorisofCHD
Liuetal.(2008) 9days Unstableangina pectoris
72;36treatedwith saffloryellow;36 treatedwith nitroglycerin (61.7±12.2)
Intravenouswith 80mgsaffloryellow onceaday
Intravenouswith 10mgnitroglycerin
PretherapyneuropepiteY levelwas
(228.5±29.8)pg/mlin saffloryellowgroup, (237.6±27.9)pg/mlin nitroglyceringroup. NeuropepiteYlevelof post-treatmentwas (149.5±24.3)pg/mlin saffloryellowgroup, (181.8±23.7)pg/mlin nitroglyceringroup
Nonereported NeuropepiteY couldbeaindexin observingchange ofunstableangina pectoris.Safflor yellowcould improveoxygen supplyin myocardium
Miaoetal. (2010)
14days CHDwith heart-blood stagnation syndrome
439;330intestgroup; 109incontrolgroup (18–65)
Testgroupreceived saffloryellowinjection 5ml(250mg)in0.9% NaCl250ml
Givensafflower injection20mlin0.9% NaCl250ml
Thetotaleffectiverateon anginapectorisand electrocardiogram analyzedbyperprotocol analysiswas91.6%and 67.3%respectivelyintest group,whichwas69.2% and61.2%respectivelyin controlgroup
Adverseevents5 patients(1.5%)intest groupwhilenoadverse incontrolgroup
Saffloryellow injectionatadose of5mliseffective andsafeforthe treatmentof anginapectoris withheart-blood stagnation syndromein patientswithCHD
Liuetal. (2011a)
3weeks CHD 127,62ingroup treatedwithsafflor yellow.65ingroup treatedwithusual westernmedicine (68.58±10.12)
Intravenouswith 80mgsaffloryellow 100mg/day
Usualwestern medicinetreatment
Saffloryellowtreatment resultedindecreaseof plasmaofendothelin, matrix
metalloproteinase-9and highsensitivityCreactive protein(hs-CRP)asusual treatment
Nonereported Saffloryellowis beneficialtoCHD
Liao(2012) 30days Chronicpulmonary heartdisease (CPHD)
68,34pergroup (58.78±1.74)
Usualtreatmentwith injectionof30ml safflowerinjectionin 0.5%glucoseinjection 250mlonceaday
Usualtreatment Theparametersofblood gasintheobservation groupweresignificantly betterthanthecontrol group(p<0.05)
Nonereported Safflowerinjection hasclinicalvaluein CPHD
Chenetal. (2014)
10days CPHD 72,36pergroup (58±7)
Usualtreatmentwith 100mgsaffloryellow in0.9%NaCl250ml, 30–40drop/min
Usualtreatment(7ml 10%KCl,4IUinsulin, ATP,CoAandsoonin 0.9%NaCl250ml
TheimprovementofST segmentdepressionin treatmentgroupwasmuch moreobviousthanthatin controlgroup.Thelevelof troponin,phosphocreatine kinaseandhs-CRP decreasedmuchmorethan thatincontrolgroup
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Chart2(Continued)
Author(Date) Designduration Condition No.ofparticipants
randomized(age)
Intervention Comparison Mainresultsbetween
groups
Adverseevents Authorsconclusion
RenandMei (2014)
14days Chronicobstructive pulmonarydisease (COPD)
69,35intreatedgroup, 34incontrolgroup (63.8±5.3)
Usualtreatmentand injecting150mgsafflor yellowpowderin0.9% NaCl250mlonceaday
Usualtreatment TheN-terminal pro-brainnatriuretic (NT-proBNP)was reducedto 438.39pg/mlfrom 2335.38intreated groupwhileitwas reducedto 738.27pg/mlfrom 2178.35incontrol group.
Hemorheologyindexes andpulmonary ventilationfunction wereimproved
Nonereported Saffloryellowpigment injectioncombined withwesternmedicine canbeneficialtoCOPD
Luoetal. (2014)
14days Acutecerebral infarction(ACI)
51,25intreatedgroup, 27incontrolgroup (35–40)
Intravenouswith safflowerinjection (20ml/day)and20% mannitol,citicoline, takenaspirinorally
Injectedwith20% mannitol,citicoline andtakenaspirinorally
On14thday, neurologicalseverity scores(NSS)of treatmentgroupwas lowerthanthatof controlgroup (p=0.040).Also,itwas resultedinserumIL-6 leveldecreasedand IL-10levelincreased. Positivecorrelation wasobservedbetween IL-6/IL-10valueand NSSintreatmentgroup (r=0.997,p=0.048), butmotincontrol group(r=0.962,
p=0.177)
Nonereported Earlyapplicationof safflowerinjectionis beneficialforyoung patientswithACI.The mechanismmightbe relatedwithlowing IL-6/IL-10value throughdecreasing IL-6leveland enhancingIL-10level
Xingetal. (2008)
15days Ischemicapoplexy 60,30pergroup (65±3.1)
IVdripofsaffloryellow 100mgin0.9%NaCl 250ml
20mlGinkgoleaf extractand
dipyridamoleinjection in0.9%NaCl250ml
Theeffectiveratein saffloryellowgroup was93.3%while63.3% incontrolgroup. Saffloryellowshowed aremarkableeffectfor qivacuitybloodstasis
Nonereported Saffloryellowhas favorableresultsin treatmentofischemic apoplexy
Lietal.(2011) 5days Acutegouty arthritis
120,80insafflower injectiongroup,40in colchicinegroup (30–58)
IVdripofsafflower injectionin0.9%NaCl 250mlonceaday
Taken1mgcolchicine orallythreetimesaday
Blooduricacidinboth groupsdecreasedthan pretherapy
Nonereportedin safflowerinjection groupbutserious gastrointestinal reactionhappenedin 29patientsofcontrol group
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Wei(2011) 28days Diabetic nephropathy(DN)
93,47intreatedgroup, 46incontrolgroup (57.2±9.8)
IVdripofsafflower injection20mloncea dayandusual treatment
Usualtreatment Theindexoffasting serumglucose,blood ureanitrogen,serum creatinineandurine proteindecreased muchmorethan controlgroup
Nonereported Safflowerinjection couldeffectively postponementthe developmentofrenal failurecausedby diabeticnephropathy
Pangetal. (2014)
7days Renal transplantation
382,231ingroup treatedwithsafflor yellow.151ingroup withconventional treatment(38.0±13.1)
Intravenousinjection ofsaffloryellowtwice aday(10mlpertime) andconventional treatment
Conventional treatment
Theincidenceof delayedgraftfunction waslowerin observationgroup (5.63%)thanthatin controlgroup(11.26%), whichissameasthe levelofserum creatinine,therenal segmentalandlobular arteryresistance indexes
Nonereported Saffloryellowcould descendtheincidence ofdelayedgraft functionand amelioratethe recoveryofgraft functionafterrenal transplantation
Xiongand Dong(2014)
14days Diabetic retinopathy(DR)
168,92intreatment group,76incontrol group(60.45±6.10)
Conventional treatmentwith additional100ml saffloryellowinjection onceaday
Conventional treatment
Thetotaleffectiverates oftreatmentgroupand controlgroupwere 91.3%and76.32% respectively.The increasedserum endostatinand decreasedvascular endothelialgrowth factorweremuchmore intreatedgroup
Nonereported Saffloryellowinjection canbeusedtoDRin type2diabetes patientsandthereby delaytheprogression ofDR
Yangetal. (2011)
14days DN 72,38intreatment group,34incontrol group(59.0±7.7)
IVdripofsaffloryellow 100mgin0.9%NaCl 250mlonceaday
IVdripof20ml compositesalviain 0.9%NaCl250mlonce aday
24hUrineprotein serumcreatininewere decreasedbothin prophaseandclinical phasethancontrol group
Nonereported Saffloryellowinjection wasbeneficialtoDN treatment
YangandDai (2010)
Onemonth Diabeticperipheral neuropathy(DPN)
84,42pergroup (48.94±7.15)
Conventional treatmentwith injecting100mgsafflor yellowtozusanli acupointand sanyinjiaoacupoint
Conventional treatmentwith additional100mg saffloryellowinjection onceaday
Thetotaleffectiverates oftreatmentgroupand controlgroupwere 84.47%and62.46% respectively.The treatedgroupof sensorythreshold, motornerve conductionvelocity andsensory conductionvelocity outweighthatin controlgroup
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dosageof 180mg/kg for ninety days resultedin slight nephro-toxicityinSDrats(Liuetal.,2004),whereas␣-terpineolinduced fattyliver(Choietal.,2013).Aclinicalobservationalsoreported
Carthamiflosasanewcauseofoccupationalasthma,asshownby bronchialchallengesorbrochialprovocationtests(Compesetal., 2006).Asmentionedpreviously,furtherevaluationsofthesystemic toxicityandsafetyofCarthamiflosareneeded.
Conclusion
The available pharmacological studies on crude extracts or
identified compounds of Carthami flos provide pragmatic
sup-portforsometraditionaltherapeuticclaims.However,thereare anumberofissuesthatneedtobeaddressed.First,theextensive pharmacologicalinvestigationsonquinochalconeglycosideshave predominantlyfocused onHSYA.Itsadvantages inbrain tissue, myocardialtissue,diabetesmellitus,andhypertensionhavebeen reported,anddifferentpreparationshavebeenappliedinclinical practiceinChina.However,theunderlyingmolecularmechanisms ofactionofHSYAhavenotbeensufficientlyclarified.Consequently, morerigorousexperimentsoninvitroandinvivosystems,aswell asinhumanmodels,arerequired.Moreover,howtoexploitother quinochalconeglycosidesremainsasubjectofcontinuingstudy. Second,moresystemicevaluationsofCarthamiflosinclinical appli-cations,including treatment of coronary heart disease, chronic pulmonaryheartdisease,cerebrovasculardiseases,orthopedic dis-eases,anddiabetesmellitus,needtobecarriedout.Third,inspite ofthedistinguishedactivitiesofCarthamiflosinsomediseases, theside effects ofits useshouldnot beignored, suchas sper-matogeneticfailure,allergicreaction,andnephrotoxicity.Thereby, systemictoxicityandsafetyevaluationsregardingCarthamiflosare necessary.In summary,thechallengeforthefutureofCarthami flosliesinconfirmingthemechanismunderlyingitseffectsandin providingbrawinestclinicalsupport.
Author’scontribution
YHTandYRXcontributedwithdatacollectionandwritingof themanuscript.DDGcontributedwithdatacollectionandformat ofthemanuscript.LNSandMLGsuggestedthemanuscriptoutline andguidedthewritingofthemanuscriptanddataanalysis.Allthe authorscontributedtothecriticalreadingofthemanuscript.
Conflictsofinterest
Theauthorsdeclarenoconflictsofinterest.
Acknowledgements
Thisworkwassupportedbythefollowinggrants:National
Nat-uralScienceFoundationofChina(81173484,81473300)andNew
drugMajorProgram(2009ZX19105-01)forscholarshipand finan-cialsupport.
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