Kolaviron, a Garcinia biflavonoid complex ameliorates hyperglycemia-mediated hepatic injury in rats via suppression of inflammatory responses
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1 Ayepol et l. BMC Complementry nd Alterntive Medicine 213, 13:363 RESEARCH ARTICLE Open Access Kolviron, Grcini iflvonoid complex meliortes hyperglycemi-medited heptic injury in rts vi suppression of inflmmtory responses Omolol R Ayepol 1, Novel N Chegou 3, Nicole L Brooks 2 nd Oluwfemi O Oguntieju 1* Astrct Bckground: Chronic inflmmtion plys crucil role in hyperglycemi-induced liver injury. Kolviron (KV), nturl iflvonoid from Grcini kol seeds hve een shown to possess nti- inflmmtory properties which hs not een explored in dietes. To our knowledge, this is the first study to investigte the effect of KV on pro-inflmmtory proteins in the liver of dietic rts. Methods: Dietes ws induced y single intrperitonel injection of streptozotocin (STZ) (5 mg/kg) in mle Wistr rts. Kolviron (1 mg/kg) ws dministered orlly five times week for six weeks. The concentrtions of cytokines nd chemokine were mesured using Bio-plex Pro mgnetic ed-sed ssys (Bio-Rd Lortories, Hercules, USA). Plsm glucose nd serum iomrkers of liver dysfunction were nlyzed with dignostic kits in n utomted clinicl chemistry nlyzer. Insulin concentrtion ws estimted y rdioimmunossy (RIA). Result: Kolviron (1mg/kg) tretment significntly meliorted hyperglycemi nd liver dysfunction. Serum levels of heptic mrker enzymes were significntly reduced in kolviron treted dietic rts. Kolviron prevented dietes induced increse in the heptic levels of proinflmmtory cytokines; interleukin (IL)-1et, IL-6, tumour necrosis fctor (TNF-α) nd monocyte chemotctic protein (MCP-1). Conclusion: The results of this study demonstrte tht the heptoprotective effects of kolviron in dietic rts my e prtly ssocited with its modulting effect on inflmmtory responses. Keywords: Dietes, Heptic injury, Kolviron, Pro-inflmmtory cytokine, Chemotctic protein Bckground Type 1 dietes mellitus (DM) is n utoimmune disorder involving immune medited recognition of pncretic β cells y uto-rective T cells with susequent relese of pro- inflmmtory cytokines tht worsen the disese stte [1]. DM chrcterized y prolonged hyperglycemi in the postprndil nd/or fsting stte [2] results from impired insulin medited glucose metolism. Uncontrolled hyperglycemi leds to progressive development of microvsculr nd mcrovsculr complictions, cusing moridity * Correspondence: oguntiejuo@cput.c.z 1 Deprtment of Biomedicl Sciences, Cpe Peninsul, Oxidtive Stress Reserch Centre, University of Technology, Bellville, South Afric Full list of uthor informtion is ville t the end of the rticle nd mortlity in dietic ptients [3-5]. Dietes is ssocited with n incresed risk of heptic injury [6,7]. It hs een reported tht the stndrdized mortlity rte from end-stge liver disese (i.e. cirrhosis) in dietic ptients is higher thn those with crdiovsculr disese [8]. To lrge extent, the effect of hyperglycemi is medited y n elevtion in the levels of pro-inflmmtory proteins. Over-production of severl inflmmtory meditors such s growth fctors, pro-inflmmtory cytokines nd chemokines hs een documented in DM [9-11]. Type 1 DM is considered s n inflmmtory process in which significnt increse of cytokines ws found in the lood of ptients with this disese. The response of heptocytes to pro-inflmmtory cytokines promotes the expression of genes tht medite the inflmmtory 213 Ayepol et l.; licensee BioMed Centrl Ltd. This is n Open Access rticle distriuted under the terms of the Cretive Commons Attriution License ( which permits unrestricted use, distriution, nd reproduction in ny medium, provided the originl work is properly cited. The Cretive Commons Pulic Domin Dediction wiver ( pplies to the dt mde ville in this rticle, unless otherwise stted.
2 Ayepol et l. BMC Complementry nd Alterntive Medicine 213, 13:363 Pge 2 of 9 process [12]. Furthermore, incresed oxidtive stress nd chronic inflmmtion ffects insulin secretion nd sensitivity [13]. Trgeting inflmmtory meditors signling through the use of nti-inflmmtory gents could therefore improve therpeutic options for dietic liver disese, dietic compliction tht is grdully gining recognition. Bitter kol (Grcini kol) elongs to the fmily of plnts clled Guttifere nd the genus Grcini. Grcini kol seeds hve een shown to contin complex mixture of polyphenolic compounds, iflvonoids, prenylted enzophenones nd xnthones which ccount for the mjority of its effects [14,15]. Kolviron (KV) is n extrct from the seeds of Grcini kol, contining complex mixture of iflvonoids nd polyphenols [16]. A numer of studies hve confirmed the ntioxidtive nd nti-inflmmtory effects of kolviron in chemiclly-induced toxicity, niml models of diseses nd in cell culture [17-2]. Although the glucose lowering effect of kolviron hs een reported in niml models of dietes mellitus [21,22], no study hs ddressed the effect of KV on inflmmtory iomrkers in dietes. In the present study, we investigted the effects of kolviron in modulting inflmmtory responses in the liver of streptozotocin-induced dietic rts. Methods Plnt mterils Fresh seeds of Grcini kol were purchsed from Bodij mrket in Idn, Oyo Stte, Nigeri nd uthenticted y Professor E. A Ayodele t the Deprtment of Botny, University of Idn. A voucher specimen is ville t the herrium of the Forestry Reserch Institute of Nigeri (FRIN), Idn. Extrction of kolviron Grcini kol seeds were peeled, sliced nd ir-dried (25 28 C). Kolviron ws isolted ccording to the method of Iwu et l. [2]. Briefly, the powdered seeds were extrcted with light petroleum ether (p 4 6 C) in soxhlet for 24 hr. The deftted dried product ws repcked nd extrcted with cetone. The extrct ws concentrted nd diluted twice its volume with wter nd extrcted with ethylcette (6 3 ml). The concentrted ethylcette yielded kolviron, golden yellow solid [16]. Ethics sttement The study protocol ws pproved y the Fculty of Helth nd Wellness Sciences Reserch Ethics Committee of Cpe Peninsul University of Technology (Ethics Certificte no: CPUT/HW-REC 212/AO4). All the nimls received humne cre in ccordnce to the criteri outlined in the Guide for the Cre nd Use of Lortory Animls prepred y the Ntionl Acdemy of Science (NAS) nd pulished y the Ntionl Institute of Helth (Puliction no. 8 23, revised 1978). Animls Adult mle Wistr rts, weighing out g were housed in individul plstic cges t the niml fcility of the Medicl Reserch Council, South Afric. They were supplied with wter nd stndrd rt feed d liitum. The nimls were mintined under stndrd lortory conditions t 22 ± 2 C with 12-h light/drk cycles nd humidity t 55 ± 5%. Induction of dietes Dietes ws induced in overnight fsted rts y single intrperitonel injection of freshly prepred solution of streptozotocin (STZ, Sigm, USA) in citrte uffer (.1 M, ph 4.5) t dosge of 5 mg/kg ody weight (.wt.). Dietes ws confirmed y stle hyperglycemi (>18 mmol/l) in the til lood glucose fter five dys of STZ injection using portle glucometer (Accu-Chek, Roche, Germny). Study design The dose of kolviron (1 mg/kg) ws chosen sed on our preliminry investigtion. 1 mg/kg kolviron ws more effective dose mong the doses (1 nd 2 mg/kg) investigted in our preliminry study. The nimls were divided into 4 groups (n = 1 per group): Norml control (NC group), Kolviron treted norml control (NC + KV), dietic control (DM group), nd kolviron treteddietic group (KV + DM group). Kolviron (1 mg/kg. wt.), dissolved in dimethylsulphoxide (DMSO) ws dministered y gstric gvge 5 times week. Control rts lso received DMSO s vehicle. At the end of the tretment period, the rts were weighed nd then nesthetized with n intrperitonel injection of sodium pentoritl (6 mg/kg). Blood glucose ws mesured in 4 hours-fsted nimls (usully etween 1 m nd 2 pm). Blood smples were collected from the dominl ort into glucose tues (contining sodium fluoride/potssium oxlte), EDTA-contining tues nd serum clot ctivtor tues. Blood smples were centrifuged t 4 g for 1 min t 4 C. Aliquot of the superntnt ws stored t - 8 C for plsm glucose determintion while other iochemicl nlysis ws crried out on the serum. The liver ws dissected out, rinsed in cold phosphte uffered sline (1 mm ph 7.2), lotted on filter pper nd weighed. Liver homogente ws prepred in phosphte uffered sline (1 mm ph 7.2), centrifuged t 15 rpm for 1 min t 4 C. Liquid chromtogrphy-mss spectrometry (LC-MS) nlysis of Grcini kol seed extrct LC-MS ws performed on Dionex HPLC system (Dionex Softron, Germering, Germny) equipped with inry
3 Ayepol et l. BMC Complementry nd Alterntive Medicine 213, 13:363 Pge 3 of 9 solvent mnger nd utosmpler coupled to Brucker ESI Q-TOF mss spectrometer (Bruker Dltonik GmH, Germny). Kolviron ws seprted y reversed phse chromtogrphy on Thermo Fischer Scientific C18 column 5 μm; mm (Bellefonte, USA) using grdient elution with.1% formic cid in wter (solvent A) nd cetonitrile (solvent B) s solvent t flow rte of 1. ml min -1, n injection volume of 1 μl nd n oven temperture of 3 C. MS spectr were cquired in negtive mode using the full scn nd uto MS/MS (collision energy 25 ev) scn modes with dul spry for reference mss solution. Electrospry voltge ws set to +35 V. Dry gs flow ws set to 9 l min -1 with temperture of 3 C nd neulizer gs pressure ws set to 35 psi. Anlysis of glucose nd liver dysfunction iomrkers Plsm glucose, levels of sprtte trnsminse (AST) nd lnine trnsminse (ALT) in the serum were nlyzed with dignostic kits in n utomted clinicl chemistry nlyzer (Medicl Coopertion, Bedford, MA, USA). Insulin estimtion Plsm insulin ws estimted y rdioimmunossy (RIA) ccording to the protocol supplied y Merck Millipore (Millipore, Coopertion, MA, USA). Seprte tues contining 1 μl nd 2 μl of ssy uffer, 1 μl of plsm smples or stndrds were mixed with1 μl 125 I insulin trcer nd 1 μl of primry ntiodies. The mixture ws incuted overnight t 4 C. This ws followed y the ddition of 1 ml of precipitting gent nd incution for 2 minutes t 4 C. Agin, smples were centrifuged t 4 g for 3 min t 4 C nd the superntnt ws spirted. The tues were sujected to rdioctive counting using 125 I gmm counter. Anlysis of inflmmtory iomrkers The levels of 4 inflmmtory mrkers including interleukin (IL)-1β, IL-6, tumour necrosis fctor (TNF)-α nd monocyte chemotctic protein (MCP-1) were mesured in the tissue lystes from ll the rts. This ws done using Bio-plex Pro mgnetic ed-sed ssys (Bio- Rd Lortories, Hercules, USA) on the Bio-plex pltform (Bio-Rd), ccording to the mnufcturer s instructions. Following previous optimiztion, smples were evluted undiluted, in linded mnner. Bio-Plex Mnger softwre, version 6. ws used for ed cquisition nd nlysis. Sttisticl nlysis Dt were nlyzed using one-wy nlysis of vrince nd expressed s men ± stndrd devition. Sttisticl nlyses were performed using Grph Pd Prism version 6.2, for windows (Grph Pd softwre, Sn Diego, CA). Differences were considered significnt t P <.5. Results Kolviron tretment lowers lood glucose, prevented loss of ody weight nd liver hypertrophy in dietic rts Effect of kolviron dministrtion on lood glucose, liver nd ody weight in STZ-induced dietic nd normoglycemic rts is shown in Tle 1. Six weeks fter dietes confirmtion, the rndom lood glucose concentrtion (mmol/l) in the dietic nd control group ws ± 2.25 nd 9.93 ±.51 respectively. The lood glucose concentrtion for the norml control rts plus KV ws 8.91 ±.6 nd the dietes mellitus plus kolviron group ws ± In ddition to elevted lood glucose, dietic rts hd decresed men ody weight compred to norml control while tretment with kolviron for 6 weeks significntly lowered lood glucose nd meliorted the ody weight loss when compred to the untreted dietic group, lthough ody weight ws still significntly lower in comprison with norml control. Dietes cused n increse in reltive liver weight (expressed s % ody weight) in rts while tretment of dietic rts with kolviron significntly restored liver weight to ner norml. STZ dietic rts exhiited impired insulin relese while KV tretment of dietic rts significntly incresed plsm insulin levels compred with untreted rts. Kolviron lowers serum levels of heptic enzymes in STZ-induced dietic rts Figure 1 indictes results of kolviron dministrtion on serum levels of heptic enzymes in STZ-induced dietic rts. In the dietic group, serum levels of liver dmge iomrkers; ALT (77.96 ± 11.9) nd AST (17 ± 5.43) were elevted compred with norml controls viz: 6.37 ± 7.2 nd ± 6.63 respectively. Kolviron dministrtion to dietic rts significntly reduced serum levels of ALT (67.9 ± 6.94) nd AST (53.38 ± 4.93) when compred to dietic control. Kolviron meliorted hyperglycemi-medited increse in the levels of proinflmmtory proteins in the liver of dietic rts The effect of kolviron on interleukin (IL)-1β, IL-6, tumour necrosis fctor (TNF- α) nd monocyte chemotctic protein (MCP-1) is illustrted in Figure 2. The concentrtion of proinflmmtory cytokines were significntly incresed in the liver of dietic rts when compred with the control rts. Lowered levels of MCP-1 nd IL-1β were detected in the liver of kolviron treted dietic rts compred to the untreted dietic group. Administrtion of kolviron to dietic rts 5 times week for 6 weeks lso significntly reduced IL-6 nd TNF- α when compred with oth norml control nd dietic rts. Kolviron lso lowered serum levels of TNF-α nd IL-6 in norml rts.
4 Ayepol et l. BMC Complementry nd Alterntive Medicine 213, 13:363 Pge 4 of 9 Tle 1 Effect of kolviron dministrtion on plsm glucose, insulin, liver weight nd ody weight in STZ-induced dietic nd normoglycemic rts Prmeters NC NC + KV DM DM + KV Glucose (mmol/l) 9.93 ± ±.6, ± ±2.36, Insulin (ng/ml) 7.99 ± ± ±.13,.79 ±.28, Body weight chnge (g) , Reltive liver weight (g) 3.22 ± ± ± ±.15, Dt re presented s men ± S.D Vlues. p <.5 vs. norml control. p <.5 vs. untreted dietes. NC; Norml control, NC + KV; Norml control treted with kolviron, DM; untreted dietic rts, DM + KV; Dietic rts treted with kolviron. Liquid chromtogrphy-mss spectrometry (LC-MS) nlysis of Grcini kol seed extrct In the negtive-ion, the ESI-MS nlysis of kolviron shows moleculr ion peks [M-H] - t (1) m/z , (2) m/z , (3) m/z , (4) m/z (5) m/z nd (6) m/z (Figure 3). Bsed on the clculted moleculr formul, the presence of Grcini iflvonoid 2 (m/z = C 3 H 22 O 12 ), Grcini iflvonoid 1(m/z , = C 3 H 22 O 11 ), kolflvnone (m/z = C 31 H 24 O 12 ) nd kolflvones (m/z = C 3 H 21 O 11 ), previously reported in literture s components of kolviron [16] were confirmed (Figure 4). The mss spectrum of pek 5 shows the ion pek [M-H] - t m/z with the formul C 3 H 22 O 1. On the sis of this dt nd dtse serching, the structure of this compound (pek 5) ws deduced to e inringenin (Figure 4), iflvnone commonly found in Grcini species [23] To our knowledge, this is the first report of new iflvonoid in kolviron ( Grcini iflvonoid complex). Discussion Limittions of the currently used drugs on glycemic regultion hve rised the need for the development of new drugs which cn ct s lterntive nd/or complementry therpy. Interest in nturl plnt products s nti-dietic gents hs incresed over the yers due to their low side effects nd multidimensionl mode of ction [24]. Kolviron nturl compound from the itter kol seed, contining complex mixture of Grcini iflvonoid, GBI, GB2 nd kolflvones hs een demonstrted for its, hypoglycemic, ntioxidtive, nti-inflmmtory nd ntigenotoxic effects [19,25]. The concept tht chronic low grde inflmmtion is importnt in the development nd progression of dietes nd its ssocited compliction is not new. Prolonged hyperglycemi cn generte n inflmmtory stte leding to n increment in cytokine production nd pncretic et cell destruction [26]. Anti-inflmmtory gents hve een documented to e eneficil in dietes. Among these re curcumin nd its derivtive [27,28], resvertrol [29] nd cnnidiol [3]. The dmging effect of inflmmtory molecules in dietes cn e medited through its interction with receptors nd ctivtion of signling pthwys which excerte the disese stte [31]. Due to the existing ssocition etween chronic inflmmtion nd dietic complictions including liver injury, identifiction of therpeutic trgets tht is le to specificlly downregulte proinflmmtory responses nd meditors could e promising strtegy in the mngement of dietes mellitus. It is noteworthy tht our study is the first to investigte the effect of kolviron on inflmmtory meditors in dietes. Regultion of lood glucose either in the fsting stte nd/or postprndilly is n importnt fctor in dietic therpy. Sustined glycemic control decreses the risk of NC NC + K V DM AST (U/L) DM + KV NC N C + KV D M ALT (U/L) Figure 1 Effect of kolviron dministrtion on serum levels of heptic enzymes in dietic nd normoglycemic rts. Dt re presented s men ± S.D vlues. p <.5 vs. norml control. p <.5 vs. untreted dietes. NC; Norml control, NC + KV; Norml control treted with kolviron, DM; untreted dietic rts, DM + KV; Dietic rts treted with kolviron. D M + KV
5 Ayepol et l. BMC Complementry nd Alterntive Medicine 213, 13:363 Pge 5 of ,, N C NC + KV D M D M + KV NC N C +KV D M DM + KV MCP-1 (pg/g) TNF-lph (pg/g) 3 2, N C N C + KV DM IL-6 (pg/g) DM + KV NC N C+ KV D M il-1 et (pg/g) Figure 2 Effects of kolviron on levels of MCP-1, IL-1β, TNF-α nd IL-6 in the liver of norml nd dietic rts. Dt re presented s men pg/g wet tissue ± S.D. p <.5 vs. norml control. p <.5 vs. untreted dietes. NC; Norml control, NC + KV; Norml control treted with kolviron, DM; untreted dietic rts, DM + KV; Dietic rts treted with kolviron. D M + KV developing microvsculr nd mcrovsculr complictions [32,33]. The mrked reduction of lood glucose in this study following KV tretment is in line with previous studies demonstrting its hypoglycemic effects [34]. Although kolviron significntly incresed the plsm insulin levels in dietic rts, the mgnitude of increse is lower compred to the corresponding effect on lood glucose. The mechnisms of hypoglycemic effect of kolviron might e due to the comintion of its stimulting ction on the pncretic β cells to relese insulin nd lso n insulin independent effect nd extrpncretic ction which involves glucose utiliztion in extrheptic tissues [35,36]. Furthermore, the hypoglycemic effect of flvonoids cn e medited through n increse in heptic glucose storge y stimulting the ction of glycolytic nd glycogenic enzymes or y inhiiting glucose-6-phosphtse. This consequently results in the uptke of glucose into cells nd the reduction in the lood glucose level through the upregultion of glycogen formtion, downregultion of the rte of glycogen rekdown, nd glucose synthesis [37-39]. It hs een shown tht solute or reltive insulin deficiency coupled with decresed ATP production ccounts for low protein synthesis [4]. The decresed men ody weight in dietic rts could e n indiction of excessive rekdown of structurl proteins in n ttempt to compenste for low vilility of crohydrte s n energy source [41]. The ility of kolviron to protect ginst weight loss might e due to its glucose lowering effect. Liver hypertrophy (incresed liver weight) oserved in dietic rts my e due to hypoinsulinemiinduced incresed triglycerides ccumultion in the liver s lterntive glucose precursors since liver glycogen is usully depleted in STZ-induced dietic rts [42]. Liver weight (expressed s percentge of ody weight) ws significntly lower in kolviron treted rts. The ility of kolviron to restore liver glycogen levels my prtly explin its eneficil effect on liver hypertrophy in dietic rts. It ws reported in previous tht kolviron inhiited microsoml glucose-6-phosphtse in STZ dietic rts [34]. The inhiition of glucose-6 phosphtse y kolviron cn increse heptic glucose-6 phosphte which serves s sustrte for glycogen synthesis therey resulting in upregultion of heptic glycogen levels. Amino trnsferses, sprtte trnsminse (AST) nd lnine trnsminse (ALT) ctlyse mino trnsfer rections nd re used s mrkers of heptic injury [43]. Deleterious effect of hyperglycemi in the liver of dietic rts oserved in the present study is evidenced y serum elevtion of liver dmge iomrkers. The heptoprotective
6 Ayepol et l. BMC Complementry nd Alterntive Medicine 213, 13:363 Pge 6 of 9 Intens. x MS, min #( ) x MS, min #( ) m/z Intens. x MS, min #( ) x MS, min #( ) m/z Intens. x MS, min #( ) x MS, min #(1974-2) m/z Figure 3 Mss Spectr of kolviron. (1) Grcini iflvonoid 2 (C 3 H 22 O 12, m/z ); (2) Grcini iflvonoid 1 (C 3 H 22 O 11, m/z ); (3) Kolflvnone (C 31 H 24 O 12, m/z ); (4) Grcini iflvonoid 1 (C 3 H 22 O 11, m/z ); (5) X (C 3 H 22 O 1, ), deduced to e inringenin); (6) Kolflvone (C 3 H 21 O 11, m/z ).
7 Ayepol et l. BMC Complementry nd Alterntive Medicine 213, 13:363 Pge 7 of 9 GB1 GB1 4 effect of kolviron is demonstrted y the significnt reduction of serum levels of ALT nd AST in the dietic treted rts. Inflmmtion hs een reported to cuse direct orgn dmge in dietic rts [44,45] nd humns [46]. Incresed levels of pro-inflmmtory cytokines MCP-1, IL- 1β, IL-6, IL-18 nd TNF-α hve een reported in dietes [47,48]. In this study, upregulted levels of these proinflmtory proteins were oserved in the liver of STZinduced dietes rts. MCP-1 is chemo-ttrctnt which promotes monocyte nd mcrophge migrtion nd ctivtion t the site of injury. Over-expression of MCP-1 exerts vrious dmging effects vi incresed production of superoxide rdicls from mcrophges, relese of lysosoml enzymes, cytokines, growth fctors nd cellulr dhesion molecules [49,5]. Animl nd humn studies hve lso shown correltion etween lood nd heptic levels of MCP-1 nd the extent of inflmmtion [51,52]. Considering the role of mcrophges in perpetuting heptic inflmmtion, reduction of MCP-1 levels my e nother mechnism y which kolviron medites its protective effect in the liver of dietic rts. Heptic infiltrting mcrophges nd Kupffer cells re sources of pro-inflmmtory cytokines such s TNF-α, IL-1 nd IL-6 in the liver [53]. Our study shows upregulted levels of these inflmmtory proteins in dietic stte while kolviron tretment notly reduced heptic 4 R1 R2 R3 R4 OH H OH H OH H OH OH Kolflvone OH H OH ---- Kolflvnone OH H OCH 3 OH Figure 4 Chemicl structure of Grcini iflvonoid complex. (Kolviron) contining Grcini iflvonoid GB-1(3,4,4,5,5,7,7 - hepthydroxy-3,8 iflvnone), GB-2 (3,4,4,5,5,5,7,7 -octhydroxy-3,8 - iflvnone), nd kolflvnone (3,4,4,5,5,5,7,7 octhydroxy-4 -methoxy-3,8 -iflvnone) is confirmed (Figure 4) while inringenin (Figure 4) is presumed to e n dditionl compound in kolviron sed on ESI-MS/MS result. levels of IL-1β, IL-6 nd TNF-α in dietic rt. The suppressing effect of kolviron on serum levels of IL-1β hs een demonstrted in chemiclly-induced inflmmtion of the colon [19]. IL-1β induces the expression of vrious genes encoding oxidnts, cytokines, chemokines, growth fctors nd dhesion molecules whose promoter region re monitored through interctions with trnscription fctor, NFκB [54-56]. IL-1β inhiits β- cell function nd promotes Fs-triggered poptosis in prt y NF-κB [57]. In GK rt model of type 2 dietes, tretment with IL-1 receptor ntgonist (IL-1R) reduced islet mrna expression of numer of inflmmtory fctors which includes: IL-1β, IL-6, TNF-α, MCP-1 nd MIP-1α [58]. Possile mechnism y which kolviron elicit its liver protective nd nti-inflmmtory effect in dietic rts could e y direct reduction of mcrophge infiltrtion nd/or y repressing NF-κB ctivtion. TNF-α is one of the mjor cytokines upregulted in dietic liver which cn promote the ctivtion of NF-κB through interction with the TNF-α receptor resulting in liver inflmmtion nd poptosis [55]. The involvement of TNF in lcoholic heptitis, virl heptitis nd ischemi/ reperfusion liver injury hs lso een documented [59]. Our study reveled tht Kolviron tretment rogted hyperglycemi induced increse in the heptic concentrtion of TNF-α. There is report tht kolviron (KV) shows inhiitory ction on prostglndin E 2 nd TNF-α production in mcrophge-like cell line [6]. Kolviron lso downregultes inos nd COX-2 expression in the liver of dimethyl nitrosmine (DMN)-treted rt vi the inhiition of DNA inding ctivity of NF-κB [61]. The nti inflmmtory nd heptoprotective effect oserved in our study might e medited vi inhiition of trnscription fctor NF-κB, key regultor of inflmmtory process. Although the results of our study demonstrted the eneficil effects of kolviron on inflmmtory response nd heptic injury in the liver of dietic rts, future studies cn ddress other possile mechnisms of ction of kolviron nd the underlying moleculr trgets of Grcini iflvonoid complex in dietes. Further investigtion my lso e necessry for complete elucidtion of the structure of kolviron. Conclusion In summry, our study reveled tht kolviron tretment meliorted hyperglycemi-induced increse in the levels of proinflmmtory cytokines nd chemokine in rt s liver nd my therefore ct s useful gent in retrding the progression of dietic liver complictions. Another importnt outcome of this study is the discovery of new compound from kolviron. This new compound long with previously identified compounds of kolviron my prtly explin its eneficil effects in dietes.
8 Ayepol et l. BMC Complementry nd Alterntive Medicine 213, 13:363 Pge 8 of 9 Competing interests The uthors declre tht they hve no competing interests. Author s contriutions ORA ws responsile for the conception nd design, crried out ll experiment, performed dt nlysis nd drfted the mnuscript. NNC collorted in the ntiinflmmtory studies nd mde contriution to the revision of the mnuscript. NLB nd OOO mde contriution to the conception nd revised the mnuscript criticlly for intellectul content. All uthors red nd pproved the finl mnuscript. Acknowledgement This study ws supported y the University Reserch Fund (URF) of the Cpe Peninsul University of Technology nd the Ntionl Reserch Foundtion, South Afric (NRF) grnted to Prof O.O Oguntieju. Author detils 1 Deprtment of Biomedicl Sciences, Cpe Peninsul, Oxidtive Stress Reserch Centre, University of Technology, Bellville, South Afric. 2 Deprtment of Wellness Sciences, Cpe Peninsul University of Technology, Cpe Town, South Afric. 3 Deprtment of Biomedicl Sciences, Centre of Excellence for Biomedicl Tuerculosis Reserch nd MRC Centre for Moleculr nd Cellulr Biology, Division of Moleculr Biology nd Humn Genetics, Stellenosch University, Tygererg, South Afric. Received: 9 Septemer 213 Accepted: 11 Decemer 213 Pulished: 2 Decemer 213 References 1. Yoon J, Jun H: Autoimmune destruction of pncretic et cells. Am J Therpeut 25, 12(6): Zntt L, de Sous E, Czrolli LH, Junior AC, Pizzoltti MG, Szpognicz B, Silv FR, Brreto M: Effect of crude extrct nd frctions from Vitex megpotmic leves on hyperglycemi in lloxn-dietic rts. J Ethnophrmcol 27, 19(1): Lüscher TF, Creger MA, Beckmn JA, Cosentino F: Dietes nd vsculr disese pthophysiology, clinicl consequences, nd medicl therpy: Prt II. 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9 Ayepol et l. BMC Complementry nd Alterntive Medicine 213, 13:363 Pge 9 of Wltner-Lw ME, Wng XL, Lw BK, Hll RK, Nwno M, Grnner DK: Epiglloctechin gllte, constituent of green te, represses heptic glucose production. J Biol Chem 22, 277: Srkhil P, Rhmipour S, Fdyevtn S, Mohmmdird A, Dehghn G, Amin G: Antidietic effect of Phlomis nisodont: effects on heptic cells lipid peroxidtion nd ntioxidnt enzymes in experimentl dietes. Phrmcol Res 27, 56: Murry AJ, Anderson RE, Wtson GC, Rdd GK, Clrke K: Uncoupling proteins in humn hert. Lncet 24, 364(9447): Rvi K, Rmchndrn B, Surmnin S: Protective effect of Eugeni jmoln seed kernel on tissue ntioxidnts in streptozotocin-induced dietic rts. Biol Phrm Bull 24, 27(8): Rwi M, Mourd M, Syed A: Biochemicl nd chnges in experimentl dietes efore nd fter tretment with Mngifer indic Psidium guv extrcts. Int J Phrm Bio Sci 211, 2(2): Li X: Protective effect of Lycium rrum polyscchrides on streptozotocin-induced oxidtive stress in rts. Int J Biol Mcromol 27, 4(5): Soetikno V, Sri FR, Veerveedu PT, Thndvryn RA, Hrim M, Sukumrn V, Lkshmnn AP, Suzuki K, Kwchi H, Wtne K: Curcumin meliortes mcrophge infiltrtion y inhiiting NF-B ctivtion nd proinflmmtory cytokines in streptozotocin induced-dietic nephropthy. Nutr Metol 211, 8(1): Bnki E, Degrell P, Kiss P, Kovcs K, Kemeny A, Csnky K, Duh A, Ngy D, Toth G, Tms A: Effect of PACAP tretment on kidney morphology nd cytokine expression in rt dietic nephropthy. Peptides 213, 42: Wu C, Chen J, Lu K, Chen C, Lin S, Chu P, Sytwu H, Lin Y: Aerrnt cytokines/chemokines production correlte with proteinuri in ptients with overt dietic nephropthy. Clin Chim Act 21, 411(9): Aggrwl BB: Trgeting inflmmtion-induced oesity nd metolic diseses y curcumin nd other nutrceuticls. Annu Rev Nutr 21, 3: Jin SK, Rins J, Crod J, Lrson B, Jones K: Curcumin supplementtion lowers TNF-α, IL-6, IL-8, nd MCP-1 secretion in high glucose-treted cultured monocytes nd lood levels of TNF-α, IL-6, MCP-1, glucose, nd glycosylted hemogloin in dietic rts. Antioxid Redox Signl 29, 11(2): Viedt C, Orth SR: Monocyte chemottrctnt protein-1 (MCP 1) in the kidney: does it more thn simply ttrct monocytes? Nephrol Dil Trnsplnt 22, 17(12): Tesch GH: MCP-1/CCL2: new dignostic mrker nd therpeutic trget for progressive renl injury in dietic nephropthy. Am J Physiol Ren Physiol 28, 294(4):F697 F Tous M, Ferré N, Rull A, Mrsillch J, Coll B, Alonso-Villverde C, Cmps J, Joven J: Dietry cholesterol nd differentil monocyte chemottrctnt protein-1 gene expression in ort nd liver of po E-deficient mice. Biochem Biophys Res Commun 26, 34(4): Tmur Y, Sugimoto M, Murym T, Ued Y, Knmori H, Ono K, Ariysu H, Akmizu T, Kit T, Yokode M: Inhiition of CCR2 meliortes insulin resistnce nd heptic stetosis in d/d mice. Arterioscler Throm Vsc Biol 28, 28(12): Tilg H, Diehl AM: Cytokines in lcoholic nd nonlcoholic stetoheptitis. New Engl J Med 2, 343: Solt LA, Mdge LA, Ornge JS, My MJ: Interleukin-1-induced NF-κB ctivtion is NEMO-dependent ut does not require IKKβ. J Biol Chem 27, 282(12): Ingrmo PI, Ronco MT, Frncés DE, Monti JA, Pisni GB, Cellos MP, Glleno M, Crrillo MC, Crnovle CE: Tumor necrosis fctor lph pthwys develops liver poptosis in type 1 dietes mellitus. Mol Immunol 211, 48(12): Gui D, Hung J, Guo Y, Chen J, Chen Y, Xio W, Liu X, Wng N: Astrgloside IV meliortes renl injury in streptozotocin-induced dietic rts through inhiiting NF-κB- medited inflmmtory genes expression. Cytokine 213, 61: Cnop M, Welsh N, Jons J, Jörns A, Lenzen S, Eizirik DL: Mechnisms of pncretic β- Cell deth in type 1 nd type 2 dietes, mny differences, few Similrities. Dietes 25, 54(suppl 2):S97 S Ehses J, Lcrz G, Giroix M, Schmidlin F, Coulud J, Kssis N, Irminger J, Kergot M, Porth B, Homo-Delrche F: IL-1 ntgonism reduces hyperglycemi nd tissue inflmmtion in the type 2 dietic GK rt. Proc Ntl Acd Sci 29, 16(33): Ding W, Yin X: Dissection of the multiple mechnisms of TNF α induced poptosis in liver injury. J Cell Mol Med 24, 8(4): Olleye S, Onsnwo S, Ige A, Wu K, Cho C: Anti-inflmmtory ctivities of kolviron- inhiition of nitric oxide, prostglndin E2 nd tumor necrosis fctor-lph production in ctivted mcrophge-like cell line. Afr J Med Med Sci 21, 39: Fromi EO, Shrotriy S, Surh Y: Kolviron inhiits dimethyl nitrosmineinduced liver injury y suppressing COX-2 nd inos expression vi NF-κB nd AP-1. Life Sci 29, 84(5 6): doi:1.1186/ Cite this rticle s: Ayepol et l.: Kolviron, Grcini iflvonoid complex meliortes hyperglycemi-medited heptic injury in rts vi suppression of inflmmtory responses. BMC Complementry nd Alterntive Medicine :363. Sumit your next mnuscript to BioMed Centrl nd tke full dvntge of: Convenient online sumission Thorough peer review No spce constrints or color figure chrges Immedite puliction on cceptnce Inclusion in PuMed, CAS, Scopus nd Google Scholr Reserch which is freely ville for redistriution Sumit your mnuscript t
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