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Key words: Tazaaalo§ supetoxide
mion rsay, neuuophil md platelet
adhesion osa1s, adjuv@t dthritis,
carageenan-induced edema.
'l he oti-iaflammarory effects of
Afltid nontdflq either alone or in
combination vidr otter homeopathic
substances, have heer obseryed in
experimental and clinical studies,
Howeyer, its me.hdism of action hd
not been elucidàted. This pàper pre-
senrs the ffndings of a stu<lv of the
anri-inflammato.v effects of
Tta m.elqs (^prcpùarion ontairing
Ar ;.d tuontdfta ud other plant
extacts c vell o minerals that could
potentially work synergistically) using
rùiots in riùo and in liùÒ tuo<lels.
The etrect of Tramte"!,'S or t*o
importmt cellulù functions, nmelv
superoxide ion production ànd
human plarelet adhesion, w6 t6ted.
TtarneeFS did not affect either of
thesc celluld tun.tions, suggesting
thrt its atrti'infldmàtory effectr ùe
rot du€ to grmùloq«e ànd platel.t
iahibitioo. These lìndings also suggest
that the antimidobial fuactions of
immune cells such c granulogctes
re not &srupted by lrazrzeel.iS.
}r zao experiments using models of
chronic md acure irflammation show
thar ir the sc of subnuimal activa-
tion of the inflammatory response!
TiazzaeliS signiffcmdy rednco the
developmenr of Iocal edena Guch as
thar seen in the Èrst ph*e ofadjuvmt
arthritisl and causes a r50/o ieduction
of the carmgeenan-induced edema
*.hen adninistered locallv. These
results sugg.st that the anti-infldma-
:ory etr..ts of TtdatueelèS are not due
Coraoai.r,i EJpr n,(oraLsÌud,c on lrxumc.lls
Experimental Studies on the Anti-Inflammatory
Activity of a Homeopathic Preparation
A. CoMorri, S. Benani, tl. N,lerelnànn, S. CLirunbÒlÒ', S. Lùsignoli', and 1ì BclLalirc'
lmdrure ol lharmacology, Un;,ersLw of \ierÒnà, ltrlv
Bìologische Heiìmìrtel Hetl GnbH, Ba.Len Baden, GernàùI
'lnstittrte oI Clìnicil Che!, isrr y and lfic, oscopr: Lrniversi$ of \èiona, lrJ!
to iÌs aciion on a speciffc t?e of
imunonodùlating c€ll or due to à
biochemical mechanism associated
*ith conventional dti-infl matorv
drugs. Instea4 Ttdme.lrs àppets ro
inhibit the acute inflammatory
process at the local level.
lhe bkxsorns ot Amta nottma ate
used thempeuricilly prìnarìLv for Lhcn
ànt;inllamrnirory.nd ùrlgcsic propcr-
ries. -fhere àrc mor. tlìù 150 chcnii.al
lbrances in Arnicr blosoms rnd. of
ther. hderalir and its dcLratires and
llavoroirls are rhe most inportant. The
ùti hfl.lrmanrn' and antimic«,bial
rctìriries of thesr sub*anes have been
dcnorsrraerl bodr tu ùm tt1 it Llo
l1.ll. A nedninàl prcpanti(,n onÈin
ing rctìre substrncts frm ,.1rrirt mr
rzra rogerhe, +rth other planr ertracr
ard oirerals that could porertiallr act
srnergisd.rllv has lten Jer.elopeJ br
Heel and is callcd Trnwelt'\. rlLe prL
inI) indi.[iÒDs of ,,,r,../iJ nrdudc
vri)u\ ivp.s oflesioDs aDd nillÙuiro-
,v prucesses oI rhe nrclcs rnd jona
such as sprùx and ì:nLiscs. fts cffìcr*
has hern dernonstntcd in a nurbcL of
experimrntrl and chrì..] srudics i3,11,
incltLrlìng rardonìzcd, doublcìlind,
phcebo colnoìled oials. Horercr the
lrc.hanìsnìGl bv Bhi.h lzr,,../nJ
crcrrs ir anti inlLnnatorv eflècts hare
hererolòre ,u been .larifìeJ.
Ìr rhìs paper rve teporr ase,ies of*uJ
ics on aniiì31 models ro improre the
undesrinding of linnna!'S atul ;*
nechanim ot rnti intlamnao,y actòn
ìn comparnon rith clasicrl rnti iniìan
Consi.le.ing rh. dillìrcnr uss of
]]atnttliS iLt humus, oro rnimrl
nodeLs of inflan,narion havc bcor
ernployd. for asscsog its el1ècr on
chro c ìnflamnarlor T un,l:ts wa.
adnìliisrered ro nrs vìrh àdjùrant
anlrLtn (Ai.), rhich sLares rvirh human
rhcurmtoid arthrìris nanl ìomunologi
cal, biocliemical, and clinìcal charactcris-
rics. Adjuraor atrì,rirìs mav be induced
i! r.rs by inje.ting inb rhe hìnd paw or
rall a mrieg ol killed mycobacterla sus
penderl in ninerri oil. The sequcncc oi
phvsioìogiceL eflccrs is charactcrizerl b-r
etlv Loc nrlLunation ard, atler nvo
weeks, dìsscnnnrcd arthrirìc ìesions.
B..iusc of irs reprodu.ibilin, AA ;
widclv use,l tor rudring antl-inflamna
ron and immunomoJnlaory agens.
l,o.assesing n\ effr.t Òn r.u( nrlli!!
matx,n, 7i,,,r../is $,as Lestcd h car-
Lageenar-ìnduced rrlemr in msr a tcs
rhat is currentlv emplored for ruL{lcs of
corrcntnnal anti-in1ìammauv rinLgs.
Ìr rte caragecnan-ìnduced edema er,
rìre eanr *reìling nal have a serotonin
component since it crn be inhibited
tLsins serotonin anusÒnrs Hrta"ùìc
rloes ror rpy'ar to phv a signifìcant role
h Lhis rpc ofrcurc nflarn!ìarion, rhile
prosraglandns (l'G, flav .n ìùpotranr
rolc nr the dcvclopneri ol rhe sccond
phrse of canagccran-ìnduced foot
,\ serìes oladditional experìnents 'vas
pertormed on the basic lunctìons ofreu
rophils and platelers o invesìgrte possi
ble eiTe*s ol l/,r,,?.li,.l Òn ..ll ù
tuncrions ard po*ible crtot,xirtr
2. Mareriàls.nd methods
2.r. & zoo experiments
Adluvam aÌÌh.itn srs irdtccL{ n
I ewis rrts *eìghing lrll .190 g br nilccr
28 Biomedical Therapy / Vol. XV / No. 1 1997
CotrLni,l, &pùrucl SÌuirs on lnumel
ing ino the pLantar ufre ofthe hind
parv 0.1 n of. srspension .ontàining
0.3 nrg of heat-killed M,v.Òbrcteriu!ì
buwricum (Mb) in rnineml oil. frevious
exterinenis using a sùslension ot 0.6
mgofMb droucd slight inhibnory e1}èct
by Trdtmel6's, so we decid.d rc "se
lorver arthritogcric concentration of Mb
ln o«1er to fird the experinentJ condr
rions to better emphasizt rhis nodulato-
Ttd nelJ,S w^\ giver ì.n. (0.11
nli n0 cver rwo <1ars from dav 2 ro day
28 after arthriris ìodùcriol accordìng to
therapeuric protocol. Conrrol rats
rcceived an equJ volunie of0.9% NaCl.
Everv two days the animàls wci.
weighed ard rheir contralatcraL paw
srvelling was me*und b,r r wrteL
pledrysmomru (Éod. 7150, Uso
Ba,ìle. Milan,Italv). The parv cdcna was
*rsed bv the in.re e in pùv volune
comparcd to prv volLrme ncaured
bdorc anhrnis inductior.
fourteen.2l,18, ud 35 dai* alìer the
arlhrirogeric nrjecrion, arthrìtn delelop
ment was cvaluated bv the *me obsen,
er using an arbnrary scale as follows: Lcli
ard right hhd paws, each 0-7j lelì d.l
rìght fÒrepa§s, each 0-4.5i tuìl 0-5: eùs
0-2j nÒs. ind erÌs, er.h 0-1. The neao
parv sntllìng and thrids inder h ùc
groups of rerd ainals s,ere compand
§irh rhàt obtÀined in the coiirol group
erd rhe percentage of inl birioD rv6
Carrageenan-inductd edema was
induced br iniectlng 0.1 rnl of 1% car
ragcenan {suspended n srcrilc salinel
nìro the pLdrar surfacc of the lìind tàw
of 40 Sprague-Da§ley rats weighing
210 230 g. The lìrst group (n=21 ràrs)
receivcd 0.1 ml oi Tintùeel:.S in ù.it
hìnd pa§ one hour before the car
ragecran inleoìon od the second group
(n=8 ra§) \$ ù.àted i m. wlth 2 nicc-
dons of 0.1 ml oi araazral.S half an
houL before ad rir* cdcna inductior.
Each ofthr coorol rors (n=21) rec.n-ed
an identical \!!ùme oi 0.90,t, NaCl in
l he pr. . oLmes strc mcauLcd wit!
a nier ple-,i.soomctcr bcforc.nd 1. 3,
and i hoùi arrù -,-he càrixgccDin iljec-
tiÒr, and cdcma wa! as$sed bv measur
irg rhe j.crcase in pew volume com-
pared r'ìth rhe paw volume btfore the
inlection ol crngeenm l5l. The hern
pav welling in the treated group wrs
compared wrh thrt ofthe control group.
Thc rcsponse was *pressed s percent
ase inhibnn,n Geducrion) of nrflannu-
2.2- In litro .xpelnents
The neutrophil is one of the most
inporrut cells involved in the ;nflmr
nrtory respo,sei Prti.ùla.lY rvhen tis
suc damage or bacterial products re pre
scnt.'lheretàre. it *t o{i
the possible eilecs of laaz..l@.loD rhe
tìnuiom ofrhc reutrophil ce.Lls ìn titro-
'lìvo inportarr tunctions ol granulc
cytes verc rncarr.,l i" th. p..'.".. ."d
in the rbscnce of TidunebS, mtel'-
superolidc arion produ*n,n (vhich
reflccts the ability of the cells to nounr
an cxudarire metrbolic response nccca
sary ro kill microorganisns) ard adh*
sion to sermr coated plastic urLfìrces
(whìch nflects rhe abiliw of cclls ro
*press and actiratc mcmì:ranc adhcsìon
molecules, a piercquisirc for chcmotùìs
and phaso.rtosis). Ìr ùoLrld be poinrcd
out rhat nay conrcntioml anri nrflam-
maory md enrlgesìc compourds are
krorvn to inhibir one oL both of these
Hunar blood neutrcphils from
herlthy volunteos scre isolaed as
Jescribed prcviously I6l; upe.o*ìd.
erion ras neasured bv rhe SOD-
inhibital:le reduaion of leric,vochrone
c, and adherent ceLLs wen quantified by
measuring the monbranc cnzvmc rcid
phosphatrse 16l. 'Ìhc pcrccnragc of
adherentcelis rvas calculatcd by rncans of
a stand*d cuvc obrained wìrh knovn
numb*s ofnertropiils.
Inflanmatory and homcomric otnts
are stictLy ntcrluked ard it is stll
kro",n thar plrrelers are involved ìn
inflànrn.tor rea.tn,ns. One ìmponant
nechanism br whi.h bhrdplarele$ pd
forn thcn functions is adhqion b the
lnjured rcsclwrì1, whìch ,rarbe regard-
ed o the cmcial first sep ofrhe hom«r
ràtic piocc$. Acauired or genetic
defeos for plateLet adhesìon may serious
lv conpror sc rÌrc Ìromcosriric process,
while an umecesrrv nrcrcase ni platelet
àdhesivcnc$ may resrlr n1 an jicreÀ€d
i6k of vascula. disorders. Since
Tizaarel@.! ho been used o net ta
maric onditiom whoe the hemorhrgìc
erenrs aLe highLy probrble md snrcc
helenalin là.tones fron AJnica !ìorrana
have been found ro ìnIìbir !ìatelet
dggresàrion [7], ìr wd ol ìnterest to
erpLore platelet lincrìon ìn rhe ptesence
The adhcsion of human plateles tu
fi brinogen-coated oLfaces wo rneasurc<1
asing a caloinetìc merhod bard on
rhc mcmbrane enrme acid phosphatase
[8l. lhe p-nitrophenoL prÒduced by rhc
reactn,n was mearrecl wi$ a microplate
reÀds Àt 40t nn rganxt a plateler-fìee
b1anL. The pcrcentagc of adheLent alls
wr cJcuLrrcd or thc basis ofa sandaLd
curve obranred wih a knorvn numb* of
3. Results
The th.rapcuric administration of
I'td""Èel.S lcà. ro r significanr Leduc-
rn,n in àcurc local infÌanmation (fìnt
phae of adjuvant artÌrrftis) nr compai-
«,n b the cÒntrÒls (Fisue l). Snicc ùis
etrect occuncd durnrg thc fisr l-7 days
ìn dre hind pan shere the Mb ra,
injc.rcd and ro! durins the gener:1ized
rrthriric reacrjon of ìmmunol,glcJ ori
gin, ìt suggess a srhlbmati. à.tiÒn or
locaÌ inflammaiion mth* than r capacl
ry to nodul,te the rvhole artlritic
proce$. 'lhì"- prcmpecl us to ue drc
conventi<»a1 moJels lòr à.ute inflam-
The elkcts of 7ì&nrcl.S on.av
ngeenrn edcma rrc rcponed h !ìgure 2.
ft can be rotcd that systcmìc adnìnrisra'
tio of l/a,,,1,1is (i.m.) did not rcdrce
edena rlevdopnent. §'len the dmg was
ìoÈdeJ inro thr hind pà1v onc hour
before crrageenu, ir appcarcd to nduce
edemNoluN by up ro lrqo (p=0.05).
TÌrs nrhibirior by tiaunrbS 1s simllat
to rhc c$eci qe.red by aspirin at a dose
of 30 mg/kg ln rhe same experìmeno1
model. l.he locaì inj*ion of lir""iÈliS
rr thc sune time ard halfan hor aftcr
cdcma inductrcn did not influence thc
Biomedical Therapy / Vol. XV / No. 1 1997 29
lNouttupnìl adlesion and sùDeroridc anio.
I'XPERII{ENTAI, MOI'ELS
r:,.."g""n*i,"tu*a "r"*
Àc'r[oN oF rraralvrzz€{§
N trcphii ad6.\ion rcn{1i n,è :6iliLy oa tIÈ clls Ò dpres d r.lvte men$mr idhesion rol
cul6, x 0rre,lùisi.È là, .henoixn xnd phrgoqaosis
?lI€ler rdlìsion n dre (ùcnÌ tìrt ftp !lùc h.n!n{i. pn,r$. shi.f is {!.dr nìIe,ln,krJ rÒ
Camsesan-induc.ll .,lcsr n. m.,lèl.arùe n,nrnnnr on.
irljnanr.{hrirh n tr nìodel ol.h'onic inatrnmtrn.f,
Trb t : tianrtl'r.\ tìLitt rn erTu ittotd udt!: tt t i,rù r d I iù).
e ù si.eiifi cùn er,..!r., n,,ìihirion
Strpdonde miotr pnducrion rtt1e.6 r!.
rtspoNq lcrrsrùy ro LlÌ mi@!.!ari$s.
J by nrie.rioo in loco lh lÈroE
Fn! x.ùr! phaF !9.!U4j!Iqt! È!i!.
lbil,n oflh..dl! tu noù.r , Òrldrn. mlrboli.
I
à
j
1
5
à
Time{daF)
:.,o,o1.1,,:*
[ir 1: Àt!r.rh,Lt. al ,ì,1jtunnt /..t!s i,!A): i,j{tul prtr tulti4 ùf..1]'ir! inn ,nd ùt
t..tìttrr0 tt rn!.l t;rt.kt!'t.\ i! a,trlnLrtht:fortlq2rutL4.z8rlio)liuhti
by tt t ngltr tl lltrtlmttìt'n tùar iaù i;tli,) tit tr,4l Ò11. Dùt tnt.lrt udt r
, ,l ,Ù
il
fig.2: C,r*gatan-nlrtL e,laù: t'ar :rilmt. tf mi1! tr: tt -21). t.t: meit ng i;t
hiù pat a 2 nl oJ't'rarnc.ll.\ t hu l'?it Ln-*!rù nt=21) n ì ùt u!;Lins tùr
;ii?.iott i. . aJ 0.2 ùt .t t uwt'rs hnt tt iou btfut at di* ?ttt n in.hirn
nt-8,. Ttt. t!l^ tt u.ta ! S t) t)J nm qaùic e\Ì,ùìrun':. 'P<a 0i lsttli 1t t t?n.
LLe"elopnent oi intìanmarion drra ror
Ihc lr rr" clpcrtrnerB dulrd rh:r
/lza;rrr1-.( neirhcr srrnLrhr«1 ror
inhibite,l ùe hucriors of rlt rcr
rophiìs ùth rs rufc o{idc ifio! tnr
,lÌrrì.n :fn :JLe,l.f
FurLerrroLe. ',/i,r&t.J dìd nor
atièlt eithcr pornilch.r negrtnfh thc
plrr1t heslor sunuLtclL.r rLo nr
uiil igonìsrs, {Dìr ùd rlùonlLìlr.
A n,mm:rr ol /iz,ozrrl',.§ rct rin or
erper nenraì models nr lnz ild n, rr.,
Ih. .roq jignilì.rnl tìnL1n,s Èoor
rhes. itrlcsrlgariLrtrs is rh ar,,rir.:1.-\
,loes not 41ea, ro rcr as r comorioml
anti infìammr«,Lr drù.e ;in.€ fofe oi
the allùlu tìncrr»r rcrcd r, ,n,,
shored r.igniiìcrnt inhibition ared b"
the JnLs lrarar,lrS di.L re,lnte acrre
nLilunrLu, n dre test animLs
,",/rrl.-J', at rllc ligh.sL con.otr.rliÒn
r«rinrblc n cornccti'c rì$rcs Lrr Lo.il
ì.je.rio., k n.t roric ro l.ui.ocrrcs and
rÌarel.s. Ltiu,. rh. noDet dctcnsnr and
hoùeosriri. fLr.crioDs ot rhcsc cdls rrc
Àr nrporr.nr pÒstrnc ron.L$b! rlìrt
.a! rr. drrrf noù .tl oI ltrr g rnloc, re
crperilrcnG is rhù ,i,/,r../r-! n'.ms
\rie to, rhesc utcs oicelh ard t[crctore
rloe" not nhibi dìc anrlmi.,obiil tìrr
ddèm.s rhich the granuìocites repre-
'enr. This point appea,s paricuì: r .lg-
nitì.r.t in vies olthe f.ssihle use olnre
drug in inmunoconp,onìised p idìts
or lvhere the lu.al tn"ue rnatonu could
lead o rrl oI;rfection (i.e.. ùù,mlr
rirh LatnLiors, bum, otolrrngerl Llis
Àlorcov*, rhc j:rcr rhir Lhc ccll ncLrb
o]trm ìs noL srhullcLl ip;ncdl rs *rir
orlÙ .lrugs aqqer' rhat Lrrrrrr:J
Llocs rot hlt Lhc potentiJl, .t:ler:riùr\
clltrs olnnmno*nrLrL:ol rgo::. 1iLr
crtolrnr, oi]orornrs l4,oforsi..hl
rldesl, :nd orh:, h:.re,ial ..mtnf..r.
The use oi{ce r{enr i. rorrairJicared
in patids "irh intlamm:rion of
enrlogenru, oririr iin panirular auoin
30 Biomedical Therapv / Vol. XV / No. r 1997
il-i*i ,, ,r, a*p.i-oa s'au "' T"-"l"s
lLunlrÌ1. Ho$o€r, bAed on nÙ dJÉ
ùi. dr* nor àPPer rc be à Prohl'nr
Tte i, ,;ùo {udies con6rm thar
In,,r1./"Sdoc. not,cr rn the §me $À
* -,nreotional anti-lnJlm'mrto'J dft'g\
antl rugge* rht need to finJ new rnd
more àPP.oPflJre models m n'i g:É
,À -e.hrni'm ol,crion' 5rn* thc adlu
.:nt rrthrir. s a moJel toL rreimmune
rlrea< r. n"r nsnrfiLrntlv inhibikd bn
7iz,zc.l,S, the drug does nor àpPear ro
htr\c rmouro'uPPresitt
rmmuromodulatol e{ecB à ci N!Ù-
sio. \h,"h s c.nsistent wtrh iò !a'k Ùt
.lf.cs "n ldrkoclt. hrNooi\ /' ''e'
Hov.'er Iaaaraets! is abìe to sìgnttt-
,rndv uh,brt the 1ìr't Phrk ol rrhr'ns
whtn rhe dnot is nJtcd bv r vot'r
,p i (Mb at lo$er c"'c'ntr{roor' $
.i-,"*n.n-irdr«d eJ'm resa
r-aia1,r ,ed".e. ellemr only when
rhe irleLnoo is adrrinnrreJ ì"rll1'
,,r,,..r'"S lPPer\ o *cn ir rhsr-
otuo. effics b' rnctucting not §rd' l
i.<,fi. cell rP. Òr hrochcmic:l m(hl
;i'm amotrg ùo\e {ùdred dru' tri but
b\ lLne ,nd.omPlc( rtgularion of lure
l;-l '"flamm,tio" \hlrc n tr lnù$n
rhàt ncur"peltidù rcl''s«l b' rnsnit
,,..,. ",,di,'e' Flàl L criflcJ r'ìe l')l'
1',Jtmi,rarv- "b.e 'rro" l'd " to
hroorh.rnt rhrr IaarrazF) rnar atfe'c
"."-*",..-t..,'-'' f rnfìarrn:rt on
,*l hì«ier ,oLr.h is n*deJ to irve*r
sate rhis Po$ible lrechànism ofacrion'
Iil auzollLr L, a,'nfnrrr \, AJJmr -À'
Lssietroli s Nl.iennnàr D€ S"lJro
t Reiori G. PhamntoL Xcr 1995i 3t
(r):61 5.
tu l lleilarire ll (hLrumL'olo
nt..*,1d" C C.Lndloi G Dn P
Ldi?Lqt? Bxl .,)92, 51 .329'35
I:l s.hrodcr H, Losc[c \ Srrobrch
H.'L.."n !q vill,hn G, Till U S'l''"'
K- Thtunh. Ra. 1990t 51 :839'45 -
tsl Bel[ure I] ,{idrioli (ì' Cuz?u I
( ii;,.b.1. s, Manzrr' F' Lei c'
lnaL. Bìochatt. 1994; 216:444 5A-
l9l Brrncs l! Bdvni MG, Roges DH
fl?S 19901 11:185-89.
hrrutÒ di larna.olosir
Univcrsnr dl Verom
Ì'oiicìinico l'1. Ronn
ft"ly
S,
lll \X/rlnsmtr R, V/oti.lenbrg Hl'
q,i,.e \\ 7";6/,,,1 rtr Ìhrtui'dnfic.
r995r 1:48 62.
l7l Ktrbo I Muror H KuLo '\'
Ch";Jhu,i sK \Àn.htz Y Osu'À I
ANinicrobial aeents froF Heterorttcca
;nnlaiàes ?h"u Me,Lcn' 1994 6:ztq
,1. Mru i,
f4l Tniet w, Botho B BioL' Mtd'
l99l r 20:506.
lì Zdl l, a'rrnt( \ÙD'
P.ì,-sqke C. Fotr't MeJ'
r06:96.
ffiTÈ lh. to*trÀ,ot.l'd'aal@Ta"'i''' .r,.,,uo,. How.r.r nro,ntsr.d,er(. ,{n . no ,ra.."d:t ld
ioenrrcal rngredtens L fu'aù1t95 r!e'l rn _ ''- _. -,^.t, ",t ,hra. ,noo al d. p"
'nén,,càl ineredtenb "'' " .; ri. i... pro,iJ.ed :br+ ,co o ,l d . o"
Hypercum perfoaum, rre ar rhe Jx PurcnÒ I
6azral@stabien and ctrops are not avaitable il the U S'
1
Biomedical Therapy / VoI XV / No 1 1997 31
