Antioxidant, antidiabetic and cardioprotective activities of Dicoma anomala (sond.) used in the Basotho traditional medicine

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Date
2016
Authors
Balogun, Fatai Oladunni
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University of the Free State
Abstract in other languages 𝘚𝘤𝘳𝘰𝘭𝘭 𝘥𝘰𝘸𝘯 𝘧𝘰𝘳 𝘈𝘧𝘳𝘪𝘬𝘢𝘢𝘯𝘴, 𝘚𝘦𝘚𝘰𝘵𝘩𝘰 𝘢𝘯𝘥 𝘐𝘴𝘪𝘡𝘶𝘭𝘶
Abstract
𝑬𝒏𝒈𝒍𝒊𝒔𝒉 Dicoma anomala (Sond.) belongs to the Asteraceae family and locally called Hloenya (South Sotho), fever or stomach bush (Afrikaans). The plant is used in the management of various diseases, particularly diabetes mellitus among the Basotho tribe of eastern Free State Province, South Africa. The study evaluates the antioxidant, antidiabetic and cardioprotective potentials of the plant as a way of validating the folkloric usage. The result of in vitro antioxidant assays [2, 2- azino-bis (3-ethylbenzothiazoline-6-) sulfonic acid (ABTS), reducing power, superoxide anion, hydroxyl radicals, 1,1-diphenyl-2-picryl hydrazyl (DPPH) radicals, etc.] as well as phytochemicals (such as total phenol, total flavonoids and total antioxidant capacity) in various concentrations (1.56-25 µg/ml) tested using water, ethanol, hydro-ethanol and methanol extracts of the plant’s root revealed that the water extract exhibited the best activity with half maximal inhibitory concentration (IC50: 15.20, 11.70, and 0.84 µg/mL) in DPPH, hydroxyl radical, and superoxide anion radicals respectively. The four extracts also possessed high phenolic contents, total antioxidant capacity with lower total flavonoids content. The effect of treatment with 125, 250 and 250 mg/kg body weight (b.w.) aqueous roots extract of Dicoma anomala (AQRED) was investigated in vivo in CCl4- induced hepatotoxic rats in a 15-day curative and prophylactic study. The result revealed that pre-treatment and treatment with AQRED lowers the elevated serum activities of aspartate transaminase (AST), alanine aminotransferase (ALT) and the level of thiobarbituric acid reactive species (TBARS) while restoring the activities of liver antioxidant enzymes such as catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase (SOD) towards normal control in a dose-dependent manner. This result proved the antioxidant and hepatoprotective activity of the plant The in vitro antidiabetic potential of D. anomala was investigated via the inhibition of αamylase and α-glucosidase using same extracts (as above) at the range of 1.56 - 25.00 µg/mL concentrations. All the tested extracts of the plant were active against both enzymes, although, the most potent against α-amylase and α -glucosidase was hydro-ethanol (IC50: 9.00 µg/mL) and water (IC50: 27.41 µg/mL) respectively. Similarly, aqueous extract of the D. anomala displayed competitive and non-competitive inhibition of α -amylase and α - glucosidase respectively using Lineweaver-Burk plot. Treatment with AQRED at concentration 125, 250 and 500 mg/kg b.w. in Wistar rats reversed towards control the elevated blood glucose levels, lipid peroxidation, lipid profile, glycosylated haemoglobin and activities of gluconeogenesis enzymes, with concomitant reduction in the activities of enzymatic antioxidants, glycolytic enzymes as well as the high-density lipoprotein – cholesterol (HDL-c) brought about by streptozotocin induction. Thus, the study proved the antihyperglycaemic activity of the plant. Additionally, AQRED at 125, 250 and 500 mg/kg b.w. was evaluated for its ameliorative activity against isoproterenol (ISP) –induced cardiotoxicity in an animal model. The results from the evaluated biochemical parameters revealed significant (p< 0.05) in these parameters was observed. The data obtained indicate that the lethal dose (LD50) of AQRED is in excess of 2000 mg/kg and its oral administration for 90 days is unlikely to cause any toxic effects. In conclusion, the results from this study proved the antioxidant, antihyperglycaemic and cardioprotective potentials of AQRED. The results further validate the folkloric usage of the plant in the management of diabetes mellitus among the Basotho tribe of Eastern Free State Province, South Africa. ___________________________________________________________________
𝑨𝒇𝒓𝒊𝒌𝒂𝒂𝒏𝒔 Dicoma anomala (Sond.) behoort tot die Asteraceae-familie en word plaaslik Hloenya (Sesotho), koors- of maagbos (Afrikaans) genoem. Die plant word gebruik in die bestuur van verskeie siektes, veral diabetes mellitus onder die Basotho-stam van die Oos-Vrystaatprovinsie, Suid-Afrika. Die studie evalueer die antioksidante, antidiabetiese en kardiobeskermende potensiaal van die plant as ’n manier om die tradisionele gebruik te bekragtig. Die resultaat van in vitro anti-oksidanttoetse [2, 2-asino-bis (3-etielbensotiasolien-6-) sulfonsuur (ABTS), verminderende krag, superoksied anioon, hidroksielradikale, 1,1-difeniel-2-pikriel hidrasiel (DPPH) radikale, ens.] sowel as totale chemikalieë, fluorfenolfenoïede, sowel as totale antioksidante kapasiteit) in verskeie konsentrasies (1.56-25 µg/ml) getoets met water-, etanol-, hidro-etanol- en metanolekstrakte van die plant se wortel, het aan die lig gebring dat die waterekstrak die beste aktiwiteit vertoon het met 'n halwe maksimale inhibitiewe konsentrasie (IC50: 15.20, 11.70 en 0.84 μg / ml) in DPPH, hidroksielradikale en superoksiedanionradikale onderskeidelik. Die vier ekstrakte het ook 'n hoë fenoliese inhoud gehad, totale antioksidant kapasiteit met 'n laer totale flavonoïede inhoud. Die effek van behandeling met 125, 250 en 250 mg/kg liggaamsgewig se waterige wortelekstrak van Dicoma anomala (AQRED) is in vivo ondersoek in CCl4-geïnduseerde hepatotoksiese rotte in ’n 15-dae kuratiewe en profilaktiese studie. Die resultaat het aan die lig gebring dat voorbehandeling en behandeling met AQRED die verhoogde serumaktiwiteite van aspartaattransaminase (AST), alanienaminotransferase (ALT) en die vlak van thiobarbituursuur-reaktiewe spesies (TBARS) verlaag, terwyl dit die aktiwiteite van lewerantioksidant-ensieme soos katalase (CAT), glutathione peroksidase (GPx) en superoksied dismutase (SOD) na normale beheer op ’n dosis-afhanklike wyse herstel. Hierdie resultaat het die antioksidant en hepatobeskermende aktiwiteit van die plant bewys. Die in vitro antidiabetiese potensiaal van D. anomala is ondersoek deur die inhibisie van α-amilase en α-glukosidase deur dieselfde ekstrakte (soos hierbo) te gebruik in die reeks van 1.56 - 25.00 µg/mL-konsentrasies. Al die getoetste ekstrakte van die plant was aktief teen beide ensieme, alhoewel die sterkste teen α-amilase en α-glukosidase onderskeidelik hidro-etanol (IC50: 9.00 µg/mL) en water (IC50: 27.41 µg/mL) was. Net so is die waterige ekstrak van die D. anomala, kompeterende en nie-kompeterende inhibisie van α-amilase en α-glukosidase onderskeidelik getoon deur gebruik te maak van Lineweaver-Burk tendens. Behandeling met AQRED teen ’n konsentrasie van 125, 250 en 500 mg/kg liggaamsgewig in Wistar-rotte, het omgekeer na die beheer van die verhoogde bloedglukosevlakke, lipiedperoksidasie, lipiedprofiel, glikosileerde hemoglobien en aktiwiteite van glukoneogenese ensieme, met gepaardgaande vermindering in die aktiwiteite van ensiematiese antioksidante, glikolitiese ensieme sowel as die hoëdigtheid lipoproteïen – cholesterol (HDL-c) veroorsaak deur streptozotosien induksie. Die studie het dus die anti-hiperglukemiese aktiwiteit van die plant bewys. Daarbenewens is AQRED teen 125, 250 en 500 mg/kg liggaamsgewig geëvalueer vir sy verbeterde aktiwiteit teen isoproterenol (ISP)-geïnduseerde kardiotoksisiteit in 'n diermodel. Die resultate van die geëvalueerde biochemiese parameters het beduidend vertoon (p<0.05). Die data wat verkry is, dui daarop dat die dodelike dosis (LD50) van AQRED meer as 2000 mg/kg is en dat die mondelinge toediening daarvan vir 90 dae waarskynlik geen toksiese effekte sal veroorsaak nie. Ten slotte, die resultate van hierdie studie het die antioksidante, antihiperglukemiese en kardiobeskermende potensiaal van AQRED bewys. Die resultate bevestig verder die tradisionele gebruik van die plant in die behandeling van diabetes mellitus onder die Basotho-stam van die Oos-Vrystaatprovinsie, Suid-Afrika. ___________________________________________________________________
𝑺𝒆𝑺𝒐𝒕𝒉𝒐 Dicoma anomala (Sond.) ke le leng la maloko a Asteraceae mme lehaeng kwano re e bitsa Hlwenya (ka Sesotho), feberu kapa stomach bush. Semela sena se sebediswa ho fodisa malwetse a fapaneng, haholoholo lefu la tswekere la diabetes mellitus hara morabe wa Basotho ba phelang Freistata Botjhabela mona Afrika Borwa. Diphuputso tsena di lekanya antioxidant, antidiabetic le kgonahalo ya ho ba le cardioprotective ya semela e le tsela ya ho nnetefatsa tshebediso ya mehla ya kgale, mehla ya neanotaba. Diphetho tsa antioxidant assays e ka ntle [2, 2- azino-bis (3-ethylbenzothiazoline-6-) sulfonic acid (ABTS), ho fokotsa matla, superoxide anion, hydroxyl radicals, 1,1-diphenyl-2-picryl hydrazyl (DPPH) radicals, jwalojwalo] esita le dikhemikhale tsa phytochemicals (jwaloka bongata bohle ba phenol, bongata bohle ba flavonoids le bongata bohle ba bokgoni ba antioxidant) ka hara metswako e fapaneng (1.56-25 µg/ml) e lekilweng ka ho sebedisa metsi, ethanol, hydro-ethanol le tse ntshitsweng ho methanol ho tswa ho motso wa semela di bontshitse hore tse ntshitsweng metsing di bontsha mosebetsi o motle haholo ka halofo ya bongata bohle bo thibelang motswako (IC50: 15.20, 11.70, le 0.84 µg/mL) in DPPH, hydroxyl radical, le superoxide anion radicals ka ho latelana. Dintho tsena tse nne tse ntshitsweng, di bile di na le dikateng tse phahameng kapa tse ngata tsa phenolic, bongata ba bokgoni ba antioxidant tse nang le bongata bo fokolang ba metswako ya flavonoids. Sekgahla sa phekolo e nang le 125, 250 le 250 mg/kg ya boima ba mmele le metso ya aqueous ya Dicoma anomala (AQRED) se ile sa batlisiswa ka hare ka CCl4- ho diroto tse filweng hepatotoxic ka matsatsi a 15 a diphuputso tsa curative le prophylactic. Diphetho di sibolotse hore pele ho kalafo le nakong ya kalafo ka AQRED, hoo ho theola mesebetsi ya serum le mesebetsi ya aspartate transaminase (AST), alanine aminotransferase (ALT) esita le boemo ba thiobarbituric acid reactive species (TBARS) ebile e le ho boloka mesebetsi ya liver antioxidant enzymes e jwaloka catalase (CAT), glutathione peroxidase (GPx) le superoxide dismutase (SOD) ho ya taolong e tlwaelehileng ka mokgwa wa tshebediso ya mothamo ka mong. Sephetho sena se bontshitse mosebetsi wa antioxidant le hepatoprotective wa semela. Bokgoni ba antidiabetic ya ka ntle ya D. anomala bo ile ba batlisiswa ho ya ka thibelo ya αamylase le α-glucosidase ka ho sebedisa metswako yona eo e ntshitsweng (jwaloka ka hodimo) ka tekanyetso ya metswako ya 1.56 - 25.00 µg/mL. Dikarolwana tsohle tsa semela tse entsweng diteko di ne di le mahlahahlaha ho kgema le dienzaeme ka bobedi, le hoja tse neng di se mahlahahlaha kgahlano le α-amylase hammoho le α -glucosidase e ne e le hydro-ethanol (IC50: 9.00 µg/mL) le metsi (IC50: 27.41 µg/mL) ka ho latelana. Ka tsela e tshwanang, aqueous e ntshitsweng ho D. anomala e bontshitse thibelo e matla le ho hloka thibelo ka nako e nngwe ho α -amylase le α - glucosidase ka ho latelana ka ho sebedisa Lineweaver-Burk plot. Kalafo ka AQRED ka motswako wa 125, 250 le 500 mg/kg b.w. ho diroto tsa Wistar e fetohetse ho ya ho taolo ya maemo a phahameng a tswekere e mading, lipid peroxidation, lipid profile, glycosylated haemoglobin le mesebetsi ya gluconeogenesis enzymes, ka phokotso e dumellanang le mesebetsi ya enzymatic antioxidants, glycolytic enzymes esita le high-density lipoprotein – cholesterol (HDL-c) e tlisitsweng ke ho kenywa ha streptozotocin. Ka tsela e jwalo, diphuputso di nnetefaditse bohlwahlwa ba antihyperglycaemic ba semela. Ho tswela pele, AQRED ka 125, 250 le 500 mg/kg b.w. e ne e hlahlobilwe boleng ba yona bakeng sa bohlwahlwa ba mosebetsi wa ameliorative kgahlano le isoproterenol (ISP) –induced cardiotoxicity ho mefuta ya diphoofolo. Diphetho tse tswang ho tekolo ya boleng ba mekgahlelo ya dikhemikhale mmeleng di bontshitse ka botlalo (p< 0.05) mekgahlelong ena mme e ile ya hlokomeleha. Deitha e fumanweng e bontsha hore lethal dose (LD50) ya AQRED e ngata ho feta 2000 mg/kg esita le tsamaiso ya yona ka molomo ka matsatsi a 90 ha ho bonahale e ka baka ditlamorao tsa kotsi efe kapa efe. Ho qetella, diphetho tsa diphuputso tsena di bontsha bokgoni ba antioxidant, antihyperglycaemic le cardioprotective ba AQRED. Diphetho tsena di bile di nnetefatsa ho tswela pele ha tshebediso ya nako ya Neanotaba ya kgale ya semela bakeng sa phekolo ya lefu la tswekere hara morabe wa Basotho ba phelang Botjhabela ba Provense ya Freistata mona Afrika Borwa. ___________________________________________________________________
𝑰𝒔𝒊𝒁𝒖𝒍𝒖 I-Dicoma anomala (Sond.) ingeyomndeni wakwa-Asteraceae futhi endaweni ebizwa ngokuthi iHloenya (South Sotho), umkhuhlane noma isisu sesisu (isiBhunu). Lesi sitshalo sisetshenziswa ekwelapheni izifo ezehlukene, ikakhulukazi isifo sikashukela esizweni sabeSuthu esisempumalanga yesifundazwe saseFree State, eNingizimu Afrikha. Ucwaningo luhlola amandla e-antioxidant, antidiabetic kanye ne-cardioprotective yesitshalo njengendlela yokuqinisekisa ukusetshenziswa kwe-folkloric. Umphumela we-in vitro antioxidant assays [2, 2-azino-bis (3-ethylbenzothiazoline-6-) sulfonic acid (ABTS), amandla okunciphisa, i-superoxide anion, i-hydroxyl radicals, i-1,1-diphenyl-2-picryl hydrazyl (DPPH) ) ama-radicals, njll.] kanye namakhemikhali e-phytochemicals (njengengqikithi ye-phenol, i-flavonoid isiyonke kanye nenani eliphelele le-antioxidant) ekugxiliseni okuhlukahlukene (1.56-25 µg/ml) ahlolwe kusetshenziswa amanzi, i-ethanol, i-hydro-ethanol kanye nezikhishwe ze-methanol zempande yesitshalo. Kwembule ukuthi ukukhishwa kwamanzi kubonisa umsebenzi ongcono kakhulu onokugxiliswa okuyingxenye enkulu ye-inhibitory (IC50: 15.20, 11.70, kanye no-0.84 µg/mL) ku-DPPH, i-hydroxyl radical, ne-superoxide anion radicals ngokulandelanayo. Izingcaphuno ezine nazo zazinokuqukethwe okuphezulu kwe-phenolic, umthamo ophelele we-antioxidant enokuqukethwe okuphansi kwe-flavonoid ephelele. Umthelela wokwelashwa nge-125, 250 kanye ne-250 mg/kg isisindo somzimba (b.w.) izimpande ezimanzi ezikhishwe i-Dicoma anomala (AQRED) yaphenywa nge-vivo kumagundane e-hepatotoxic enziwe yi-CCl4 ocwaningweni lwezinsuku eziyi-15 lokwelapha kanye ne-prophylactic. Umphumela uveze ukuthi ukwelashwa kwangaphambili kanye nokwelashwa nge-AQRED kwehlisa imisebenzi ye-serum ephakeme ye-aspartate transaminase (AST), i-alanine aminotransferase (ALT) kanye nezinga lezinhlobo ze-thiobarbituric acid ezisebenzayo (TBARS) ngenkathi kubuyiselwa imisebenzi yama-enzyme e-antioxidant yesibindi njenge-catalase (CAT), glutathione peroxidase (GPx) kanye ne-superoxide dismutase (SOD) ekulawuleni okujwayelekile ngendlela encike emthamweni. Lo mphumela wafakazela umsebenzi we-antioxidant kanye ne-hepatoprotective yesitshalo. I-in vitro antidiabetic potential of D. anomala yaphenywa ngokuvinjelwa kwe-αamylase ne-α-glucosidase kusetshenziswa okukhishwe okufanayo (njengangenhla) ebangeni le-1.56 - 25.00 µg/mL lokugxila. Zonke izingcaphuno ezihloliwe zesitshalo zazisebenza ngokumelene nama-enzyme womabili, nakuba, okunamandla kakhulu ngokumelene ne-α-amylase ne-α-glucosidase kwakuyi-hydro-ethanol (IC50: 9.00 µg/mL) namanzi (IC50: 27.41 µg/mL) ngokulandelana. Ngokufanayo, ukukhishwa okunamanzi kwe-D. anomala kubonise ukuvinjelwa kokuncintisana nokungaqhudelani kwe-α-amylase kanye ne-α - glucosidase ngokulandelana kusetshenziswa itulo le-Lineweaver-Burk. Ukwelashwa nge-AQRED ekugxiliseni ku-125, 250 kanye no-500 mg/kg b.w. kumagundane e-Wistar ahlehlisiwe ekulawuleni amazinga aphezulu kashukela egazini, i-lipid peroxidation, iphrofayili ye-lipid, i-glycosylated hemoglobin kanye nemisebenzi ye-gluconeogenesis enzymes, nokuncipha okuhambisanayo kwemisebenzi yama-antioxidant enzymatic, ama-glycolytic enzyme kanye ne-high-density lipoprotein- cholesterol (HDL) -c) okulethwa ukungeniswa kwe-streptozotocin. Ngakho-ke, ucwaningo lwafakazela umsebenzi we-antihyperglycemic wesitshalo. Ukwengeza, i-AQRED ku-125, 250 kanye no-500 mg/kg b.w. yahlolwa ngomsebenzi wayo wokuvuselela ngokumelene ne-isoproterenol (ISP) -i-cardiotoxicity eyenziwe kumodeli yezilwane. Imiphumela evela kumapharamitha we-biochemical ahloliwe yembula okubalulekile (p<0.05) kulawa mapharamitha abonwa. Idatha etholiwe ibonisa ukuthi umthamo obulalayo (LD50) we-AQRED ungaphezu kuka-2000 mg/kg futhi ukuphathwa kwawo ngomlomo izinsuku ezingu-90 akunakwenzeka ukuthi kubangele noma yimiphi imiphumela enobuthi. Sengiphetha, imiphumela yalolu cwaningo yafakazela amandla e-AQRED e-antioxidant, antihyperglycemic kanye ne-cardioprotective. Imiphumela iphinde iqinisekise ukusetshenziswa kwe-folkloric kwesitshalo ekulawulweni kwesifo sikashukela phakathi kwesizwe sabeSuthu ngasempumalanga yesifundazwe iFreyistata, eNingizimu Afrikha. ___________________________________________________________________
Description
Thesis (Ph.D.(Plant Sciences))--University of the Free State, 2016
Keywords
Antihyperglycaemia, Antioxidant, Cardioprotection, Dicoma anomala, Hepatotoxicity, Haematology, Phytoconstituents, Serum enzymes, Toxicity, Wistar rats, Compositae, Traditional medicine -- South Africa -- Free State, Basotho -- South Africa -- Free State, Medicinal plants -- South Africa -- Free State
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