Scribd Logo
Загрузить

Добро пожаловать в Scribd!

  • Triphala Research Lipid Peroxidation Anemia Etc

    Загружено:

    Ershad Shafi Ahmed
    13 просмотров7 страниц
    Triphala research

    Авторское право

    © © All Rights Reserved

    Доступные форматы

    PDF или читайте онлайн в Scribd

    Этот документ был вам полезен?

    Это неприемлемый материал?

    Пожаловаться на этот документ
    Triphala research

    Авторское право:

    © All Rights Reserved
    Скачайте в формате PDF или читайте онлайн в Scribd
    Отметить как неприемлемый контент
    13 просмотров7 страниц

    Triphala Research Lipid Peroxidation Anemia Etc

    Загружено:

    Ershad Shafi Ahmed
    Triphala research

    Авторское право:

    © All Rights Reserved
    Скачайте в формате PDF или читайте онлайн в Scribd
    Отметить как неприемлемый контент
    из 7
    International Journal of Biological Chemistry 1 G): 149-15: ISSN 1819-155% (© 2007 Academie Journal Ine. 2007 ‘Therapeutic Efficacy of Indian Ayurvedic Herbal Formulation hala on Lipid Peroxidation, Antioxidant Status and Inflammatory Mediator TNF-a in Adjuvant-induced Arthritic Mice BP. Sabine and M, Rasool School of Biotechnology, Chemical and Biomedical engineering, VIT University, Vellore-632 014, Tamil Nadu, India [bstract The aim ofthis reeatch is to detrmine the prototve ellicecy of Teiphala in adjuvantinduced aris with respect to lipid peroxidation, snoxidant satus and inflaramotory modiator tumour necrosis factor-alpha (INF-0) Artis was induced by intradermal injection of Complete Frenn’s Adjuvant (0.1 mL into the right hind paw of| ‘wis albino mice Triphala (1000 mg kg * day) and Indomethacin 3 mg kg dey) were cally administred fr 8 days (fim 11th to 18th day) afer adjuvant incon. The levels oflipid poses, enn and non enzymic anoidant and mone neces ete ere smsasurotin Hood and splon of eonrl and arte animals. Inarttc animals the iid peroxidation and tumour necrosis factor. were found increased. wheteas a decrease in snsionidant levels was observed, These observed alterations Were sgniticanly modulated t9 near nonal loves by oral administration of Triphala (1000 mg ke" day~) in atti animals, The present study shows the antartvtic effect of Triphala against adjuvant- induced artic mice. Key words: Triphala, adjuvantinduced artis, ipid peroxidation, antioxidant, tumour necrosis factor. INTRODUCTION ‘Rheumatoid Artrts (RA) isa systemie autoimmune: disease that primarily presents as a chronic symmetic polyarthritis associatad with inflammation and cartilage destruction (Costello and Halverson, 2003). [tis the most commen inflammatory artis affecting approximately 1-2% of the _general population werldwide (Haris, 1994), Incidence inereases with age, with women being affected three times more than men. The pathogenesis of rheumatoid arhrits is multifactorial and recent esearch has implicated oxygen fee radicals and inflammatory eytokin tumor necrosis factor (TNF) ‘which have been found f play an important role as mediators of disse damage (Fleischmann eal, 2041), Reactive oxygen species are highly reative transient chemical species nite oxide, supercxide anion, hydrogen peroxide and hycrox! radical) with the potential to initiate celular damage (to protein, lipids et) in join tissues, especially in rheumatoid arthritis. TNF i produced abundantly bby activated macrophages in synovial tissues fiom dheumatei artis patients and by stimulating ‘mesenchymal cells such as osteoclast, fibroblasts and chondrocytes, which release joint-destuctive matrix proteinase. Although a nunber of drags (non-steroidal or sterol antiinflammatory agents ad mmunosuppressant) used in the treatment of sheumatoid arthritis have been developed over the past few decades, there is sll an urgent need for more effective drugs with lower side effects Thus, a ‘Corresponding Aue! 134 Rasool Facly ofedial Bochansiy, Schoo eBiaedawkw, (Chmical und Bereta Engine, VIT Universe, Vellore Tal Na Indi 2014, Inka 91-1162248052, 240411 49 Int. Biol. Chem, 1 (3): 149-150, 2007 lsirable drug fo treatment of rheumatoid arthritis, which possesses the properties of controlling joint inflammation, relieving joint pain and protecting the artic joints fiom damage without any side effets is required. ‘Triphala is the mast commonly used Indian ayuaveda herbal formation, comprising the futs oftoe tees lndian goose berry (Emblica ofcinals Gaetn family-Euphobiaceae), Bellnicemsrobalan (Ferminaliabeteriea Linn fanily-Combretaceae) and Chabalic myrobalan Terminal chebula Ret, famuly-Combetoeae), Triphala has been reported to bea rich source of Vitamin C ellapie avi, gallie acid, chebulini acid, bellencanin, f-sitostrol and Havancids (Keut f a, 2005), Its components Embiiea fftcnals,Terminalia belertca, Tenminala chebula are reported to posses anfinllanvatory, antimwtagenie, antioxidant, cytoprotective, gastroprotective activity, myocardial necrosis, hepatoprotective, antibacterial, hypolipdemic and anticancer activity Pulok Mukherjee er a, 2006, ‘Siikumar er al ,2005, Sanaa ew, 2006, Saravana et a, 2007). preliminary studies confirm its antiinflammatory and lysosomal membrane stsbilizing effect on adjuvant-inuced arthritis in mice ‘Rascol and Sabina, 2007). Inthe current study, the ant-arthite effect of Triphala was investigated using adjuvant induced arti, a well known experimental model fo beumatid aritisin ice. The anke-artsitie effect of Triphala was assessed by measuring changes ia lipid petosidation, aioxidant ‘Status and inflammatory mediator Tumour Nectesis Factor (TNF) in plasma and splocn of arthritic animals. The standard noresteroidal anf-inlaramatcey drug, Indomeliin, was used asa reference dug foe purposes of comparison, MATERIALS AND METHODS. Animals ‘The study was perfonmed with Swiss albino mice, 25-30 , of either sex. The mice were brought fiom Tamil Nadu Veterinary College, Chennai, Inds. The mice were aclimatized fora week in light, ‘and temperature-contrlled room with a 12h dak-lighteyele. The mice were fed commercial pelleted fed flom Hindustan Lever Lid. (Mumbai, Inds) ancl water was made freely available, Drug ‘The commercially available Triphala powder ( mixture of diied and powdered fits of dee plants 7. chebula, 2. oficinalisandT.bellericain equal proposticns (1:1) vas obtained from Indian ‘medical Practitioners Co-operative Stores and Society (MCOPS), Adyar, Chermai, India and its aquoous suspension in % gum acacia was used at adose 1g kg™' b.wt, orally. The Triphala powder used in this study was found to contain approximately 50% polyphenols as investigated by High Performance Thin Layer Chromatography (HPTLC) densitometer analysis (Rascal and Sabina, 2007) Indornettin (Taina Dacha Pharmaceuticals, Chennai, Indi) was dissolved in 2% gum acacia solution and 3 g kg” bret. was aciministerad orally (Rasoo! and Varalakshmi, 2007), All other reagents used were standatd laboratory eazens of analytical grade and were purchased locally Dosage Preliminary studies with different dosages (250 mg, 300 mg. 1 g) of this drug revealed that 1 ekg bon. dosage produced significant ant-inflamamatory effect by redueing paw swelling in adjuwant-induoed arate animal. Hence | g kz! bat. dosage was consideted for this study. Experimental Protocol ‘The mice were divided into six groups each comprising six animals, Group 1 served as controls [mn Group Il arthritis was induced by intra dermal injction of Complets Freund's Adjuvant (CFA) (0.1 mL) into the sight hind paw (Rasool and Sabina, 2007), The adjuvant (Tubereulosis Research 130 Int. Biol. Chem, 1 (3): 149-150, 2007 (Center, Chennai, India) contained heat-killed Mycobacterium nubercufosis(H37 RA (10mg) i paraffin oil (1 mi), Group II and IV were treated with Triphala and Indomethacin, respectively, for 8 days fiom the L1th tothe 18th day ofthe experimental period. Groups V and VI comprised artitic mice Were treated with Triphala end Indomethacin, respectively, fiom day 11th to 18th afler the administration of Complete Freund's Adjuvant (CPA), (On the 19th day, at the end ofthe experimental period, the animals were sacrificed by cervical decapitation and the blood was collected. The spleen was immediately dissected out and homogenized inicecold 0.01 M, Tris HCL buff, PH7.4 fo give a 10% homogenate. Spleen tissue homogenate and plasma were used for assoving the following biochemical investigations. Biochemical Estimations Lipid peroxidation in plasma was estimated by the method of Ledwozyw et af. (1986). Spleen lipid perosiation was carved out Ly the procedure of Hogberg etal. (1974) using thiobarbutric weid (TBA) asthe colouring agent. Malonaldelyde (MDA) produced dusing peroxidation of lipids served san index ofipid peroxidation. mallonaldchiyde reacts with thiobarbutric acd to generate a colour rodiuet, which absorbs at 532 nm, Spleen lipid peroxidation with induoer system namely. 10 mM FeS0/0.2 ascorbate!10 mM 1,0, was measused by the method of Devasagayatn and Tarachand (1987), The Malonaldely/de contents of the samples were expressed as mmoles of MDA formed mproei. ‘Superoxide dismutase (SOD) activity in sploen was determined by the method of Markland and Marklund (1974), The degree of inhibition ofthe auto-oxidation of pyregallol at an alkaline pH by ‘SOD was used asa measure of the enzyme activity. Catalase and lutathione peroxidase activities in spleen were estimaled by the method of Sinha (1972) and Rotruk er al. (1973) The activity of catalase was expressed as jig of H,O, consumodiinn'mg protein Glutathione peroxidase was expressed as jg ‘of ghutathione lized minute/mg’ protein. Non enzymic antioxidants, phtathione (Moron etal. 1973), ascorbic acid (Omaye ef al, 1979), total suphycryl (TSH) and non protein sulphydyl (NPSH) (Sedlack and Lindsay, 1968) were estimated inthe spleen, ‘TNFa levels in plasma nd spleen of cntsol and arlsitie mice were detemnined by enzyme- linked immunosorbent assay (ELISA, Cayman Chemicals Company, USA), according to the -mannfactrer’s instruction. ‘Statistical Analysts Ress were expressed as meany+SD and statistical analysis was performed using ANOVA to determine significant differences between groups, followed by Student's Newman-Keuls test, p=0.05 implied sigraticance RESULTS. Ingroup Il arthritic mice, basal lipid peroxide levels wer elevated in both plasma and spleen. In presence of indueers like ascorbate, FeSO, and H,0,, spleen lipid peroxide levels were found to be ‘increased significantly compared to consol group, whereas the administration of Triphala to artstie mice altered the above changes by regulating the lipid peroxide level to nearly that of normal levels (Table 1) ‘The enzymic and non-enzymic antioxidants wore significantly decreased in arthritic mice compared to contol mice. These changes highlight the deteriorating antioxidant statu in the antrtic sce. Furthermore, administration of Tsiphula increased the enzymic and non-enzymie antioxidant levels in arthritic mice considerably, which indicates its antiperoxidative action (Table 2), rey Int. Biol. Chem, 1 (3): 149-150, 2007 ‘obleL: Blet of Hphah and lndamebncin cebasal and indice pid proidation n plasma aspen of eovl and perenne Grup V Gray WE Group! Greupl Group Gregp Vis Antti Pamsters __Cawrol_arviis_Tials_ndenhacin “Tptala_indac Plena 2IRLOIS — SASi0.3te" 2.70016 2724054 —_SOK0.1RNEE 32040208 Spleenbael 245016 S7540a4ge 254018 gDO Ie 3540 IBY TSN TINE FeSOvinbced — 3.100023 $s0unamar 3251025 ANeinIs Sein 3o—rbY 5000 Tarb+ Avcabate died 328:020 @sododmer 3ISIOIR Game AmHOauarbY L380 2% 11, ned S018 41040729" 2.604030 250013 28510 IaH $1040 178 ‘Treatment of soups: Group 1Cenkok Geayp WeArtrtie nice Grup UlCaual mice wed wit) Tephala 1000 mg Ke bo fe Say Ho 1 dh 3th dy: Grey TV-Contl ie ete wilh Inderetaes Sha St) fe ey rr 11 ht 18 dy Group VA ce teed wh Up (1000 maka 9 9) for 11 th to 18h day pos aunt Creap VL-Artate mice wetd wih Indneacs (ig kg” bw Sr 11 18 dy post aduvanl. Comparson: A-Geaup {vs groups I IV, V an VL. trop il vs group Vand VL Each clue Forel meat in Valuers egresed anoles lone omnes poten plea dL" "The pbs represent atta signee ot" p05 ‘able 2: ft of gala and Indomethacin ca the enya an one toidat eve in splen of eorland mal ce EEE Ee EEE TE Grp V Gray WT Grp Greupl!——Grep tt Grp artis Ante Panmure ___Coatel Avis ‘Trg nanan ‘Tiphala dancin Siperoide ddnmgase($0D) 1501010 OS0nGSeY 1.00.10 LS$s032LStunaTbY 1. eoRazetOY ‘Guaatene Dpernidwe (Op) 3204018 254016" 3355018 328020 Sasenaterbs 3.192019 Casie 1350129 Lososnseme 15761 11 1SAOLS A4IHEL OME 15 7OHL OS™ Ghatine L7qio0s “1.28nom "1.8040 11 L70sNS GND Las bY 1.01000 “oa spd asin T5086 T7SD4s 7651058 GND T1OLOATH onatein Lgeioge L.7s0aibe 7a ‘Treatment of gps Group T Cael, Croup = Grasp TI Control mice ete eth Tria 1000 mg Eg ba for 8 dye ro 1 dt 1h day. Grup 1V-Conl mice reed wit Indometbeat 3g a ‘avo for days fom 11 do Tt day Group V- Artie mice wat wilh Trial 1000 mag gb) Tn 1 Us to 1h ay: Group ULAMai mice eed val Indamatae i (Sg ka" wt ten 11th to 188 day. Canparisons A ‘up vs peu TV, Vand VI ab Gro I sroupV ae VL Vas re expressed a meanSD (1 ~ 6) "Eye unite we egresed ag, SOD-untsing poten (iit amouat of eye eure to bt Be ao-oe ion reaction ty M0. Gps of GSH uilizedninina peti Caleenel of HO, censaned min! mg pre ‘Slutabione, Teal suiydyt (TSE, Nen-preein slphyay (NPSHD, Vitamin C- yg mg pein. The symbol epresel tel sifeanceat * p08 sulpsdyl OSE 1880.04 TNPabpte pel seugagaz ‘Grp ” Crap" Group Group’ Grup V"Czowp VI Fig, I: Effect of Triphala and Indomethacin on TNF-alpha production in adjwvant-indwoed antrtis im mice, Treatment of groups: Group 1-Contro; Group I-Arttitic mice; Group [II-Contro ‘mice weated wih Triphala (1000 mg kg“ b.wt) for 8 days from 11 ts 0 18th day; Group IV- Control mies trated with Indomethacin 3 mg kg~* bast) foc & day from 11 th to 18th day, Group V-Arthritie mice treated with Triphala (1000 mg kg~' bawt) from 11 th to 18th day post adjuvant; Group V-Arthrtic mice treated with Indomethacin (3 mg kg bet) from 1] thto 18th day post adjuvant, Comparisons: a: Group I vs groups I, Il, IV, V and VI and b Group II vs group V and VI. The symbols represent statistical significance at: *p<0.05 132 Int. Biol. Chem, 1 (3): 149-150, 2007 Levels of tumour neerosis facto. in the arthritic mice were systemically overproduced inthe serum and sploen, while the elevated levels of taicur necrosis fator-x wre found to be decreased in ‘Triphala treated arthritic mice (Fig 1, DISCUSSION ‘Triphala an Indian ayurvedic herbal formulation is believed to promote health, immunity and longevity (Sandhya ofa, 2006), Tritt, rch in antioxidants, plays an essential rol in the treatment ‘oft wide varicty of conditions like infections, obesity, anemia, fatigue, constipation and in infectious diseases like tubsteuloss, pneumonia and AIDs (El-Mekkaneey and Merelhy, 1995). Here, we have ‘evaluated th ant-arite efTect of Triphala in adjuvantinduced artis in mice; an experimental ‘model of arthritis, which resembles several aspects of human sheumateid atts (Pearson, 1964). In rheumatoid arthritis, the polymorphomuclear leakooyles are activated and reecive oxygen species are generated in excessive amounts, These are reactive ephemeral molecules which are known toplay an important roe in the progression ofinlammmtion, In this study, the lipid peroxide level was found to be ineressed in attic mice compared to contol animals. This is in agreement with other studies in which higher lipid peroxide levels have ben reported in adjuvantinduced. arthritis (Geetha eral, 1998; Rasool and Varalukshani, 2007), The increased lipid peroxide observed in atic animals may be die to its selease ftom neutrophils and monoeytes during inflammation. I 8 an accepted fact that sheumatoid artis is accompanied by abnormalities in body iron metabolism. At the onset of inflammation, it was observed that there was @ rapid fll in the total iron content of blood plasma fllowed by an increased deposition of ron proteins in the synovial fluid. The drop in plasma iron correlates closely with the activity of the inflammatory process (Gutteridge, 1986), In the synovial fluid of inflamed joints, the iron released during nsctosis, might catalyze the formation of hydroxyl radicals fromm H,0, Uns contbuaing to un inevease in iflanamation In the present study, te concentration of enzymic and non-enzymie antioxidants was significantly decreased in athitie mice (Group ID as compared te normal control mice. Similar resalts have been reported by Geetha eral (1998) and Ramprasad etal. 2005). The increased rate of fee rial production frequonty ress ina decrease inthe level of antionidant enzymes and the enzyme Aactvitios are reduced thereby leading to autceatalysis of oxidative damage process, Super oxide ismutas, glutathione peroxidase and catalase are considered primary antcnidant enzymes, since they are involved in dzec elimination offre radicals: The decreased activity of enzyme antioxidant in ‘adjvant-induced aris may be due tothe hi concentration of eacive oxygen species formed. Due to increased lipid peroxidation, the levels of free radicals overcome the saturation level. The high concentrations of fize radicals inhibit the activity of aniioxidants and hence the activities of these ‘enzymes appear to be reduced, Enzyme antioxidants ae inactivate by the fee radicals and hence the presence of non-enzymic antioxidants is presumably essential forthe removal of these radicals (Allen, 1991), Vitamin, glutathione and oer thiols are water soluble antioxidants that remove free radial fiom eytosol by reacting direely with thera (Allen, 1991). Inthe present cbservation, non-enzymic antioxidants ere found to be significantly decreased in arthutc animals. The cbserved aon-enaymie antioxidants depression in adjuvant-induved arthritis is associated with the protracted inammatory pluse of the disease, In present study, it was observed that Triphala protects the arthite animal from lipid _proxidation and inereases the enzyme activites of superoxide dismutase, glutathione peroxidase, calalase and non-enzymife antioxidant levels, which suggests that Triphala prevents the oxidative damage due to reactive cxygen species overpoductin from artis. The increased aetvitiewlevels of emioxidants observed in teated rats could be due to phenolic compounds (lavonoids) present in ‘Triphala. Ithas been reported earlier that phenolie compounds his potetial antioxidant activity and iteould scavenge fee radicals effectively and tus also been found to intit the lipid peroxidation 133, Int. Biol. Chem, 1 (3): 149-150, 2007 (Ramprasad of af, 2005), Hycrogen-clonatng hyeroxyl groups on the aromatic ring of phenolic ‘compounds present in Triphale might be responsibe forthe free racical scavenging and antioxidant activity of phenolics, which correlates with easier report (Anand, 1997). Emblica offcinalts, Teyminaliabelerica an Terminalia chebul, the constituents of Triphala exact have been reported tobe a sich sows of Vitamin C, ellgic aid, alli aid, chebuinic ci, alericain, Psitosterol and flavanoids. Most of these compounds have bosn reported tobe a potent inbiter of ip peroxide formation, a scavenger of hydrox and supercxide radials and to inorease the antcxidant enzymes (Cagetia etal, 2002). Thus the modkving role of Triphala extract observed in oar study may be de tothe anipercnidative action ofits comps that was reported earlier (Lee eral, 2005; Naik et a, 2005) "TNF-«, a proinlammaiory cytokine, is produced by activated macrophages and other cell types and these eall types are abundant in the artic joins as has bezn shown in both the animal models. and in sheumatoid patents. Ths abundance of TNF-« in arthritic joint provides evidence of is inwolvementin de disease pathology, whichis suppevted by studies demonstrating that neutralization ‘of TNF-« leads to decreased production of cher inflammatory eytokines (Haworth ef af, 1991; Brennan ea, 1989), Present reslts confirm that Triphala suppressed the illammatory process by ‘reducing the production of tumour nectoss factor in adjuvat-induoed attic mice. ‘To conclude, the results of the present study have empirically indicate that Triphala is efective inte tetment of ceumatid arthritis and that it sappot the common belie prevailing in trations rmodicines, worldnid, REFERENCES, Allen, RG, 1991, Oxygen-esctive spevies and atiosidant respons dking development: The thetabole paradox by cellular dilferentaton Proc. Soe. Exp. Bilal. Med. 196: 117-129, Anand KKB, Singh, AK. Saxe, BK, Chandan, V.N. Gupta and V. Bhardwaj, 1997. 3, 4, ‘STiihydrcaybetenie acid (llc acid) the hepatprctctive principle inthe its of erminalia beortca bioassay guided activity, Pharmacol. Res, 36: 315-321 Brenan, FM. D. Chatry, A, Jackson, RN. Main and M. Feldman, 1989, Iniitery effect of "TNE alpha antibodies on synovial cell interleukin] production in sheumateid arthritis, Lancet, 2 nda Costello, LC. ad PR, Halverson, 2003. A new ea in sheumtoid atts weatment. Wise. Med, 102: 29.33 Devasagayam, TA and U, Taracand, 1987. Deceased lipid peroxidation in rat kidney dung gastation. Biochem. Biophys. Res. Commun., 145: 134-138. EL-Meklawey, M. and M. Merely, 1995, Inibitory effets of egyptian folk medicines on Hunan immuncetcincy virus (IV) reverse transcriptase. Chem. Pham Bull, 48: 681-648, Fleischmann, RR. Stom and I, Il, 2004. Anakin An inibitorof TL for tho treatment. of shaumatid arthritis. Exp. Opia. Biol. Therapy, 4 1333-1344 Geetha, T., P, Varalakshmi and R. Marylatha, 1998. Effect of triterpenes from Crateva nurvala stern turk on ipl petoidation in adjuvant induced ans ats, Phmaccl Res 37: 191-195 Gutteridge, J.M.C., 1986, Antioxidant properties of the prtotein ceruloplasmin, albumin and Wansfetn. A study ofthe activity in serum and synovial uid fom patients with eumatoid aris. Biochem. Biophys. Acta, 869. 119-27 Haris, E.D, 1994. Biiclogy and Pathogenesis of Rheumatoid Anhis. In; Textbook of Rhoumstology. Kelly, WN., ED. Hamris,S, Ruddy and CB, Sledge (EAs), Saunters, Piilacephia, pp: 833-873 Haworth CFM, Brennan, D, Chantry, M. Tuner, RN, Main’ and M, Feldman, 1991. Expression of gramulocyte-macrophage colony-stimulating factor in theumatoid arthritis: Regulation by tumor terns tora. EJ. fmunol, 21: 2575-2579, st Int. Biol. Chem, 1 (3): 149-150, 2007 Hogberg, 1, RE. Larson, A. Kristoferson and S. Onreius, 1974, NADPH-

    Вам также может понравиться