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Browsing by Author "Ogbe, Abdulazeez Adeola."

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    Elicitation, metabolomic analysis, and identification of antidiabetic compounds from selected indigenous plants = Ukuvuselelwa, Uhlaziyo Lokugayeka, nokuhlonzwa Kwezingxube Zesinqindasifo Sikashukela Ethathwe Ezihlahleni Zendabuko Eziqokiwe.
    (2022) Ogbe, Abdulazeez Adeola.; Van Staden, Johannes.; Finnie, Jeffrey Franklin.
    Diabetes mellitus (DM) is an endocrine disorder associated with high blood glucose levels accompanied by disruptions in the metabolism of fat, proteins and carbohydrates. DM is a chronic, non-communicable and medically incurable disease affecting millions of people globally, resulting in high morbidity and mortality rates, especially with the lingering coronavirus disease of 2019 (Covid-19). The use of western antidiabetic medicine has posed many challenges due to their perceived overall safety, treatment failure and cost. Many African communities rely on medicinal plants and their bioactive compounds as sources of medicine as a consequence of the poor state of health facilities, shortage of medical doctors and unaffordability of treatments. For this reason, this study partly evaluated the phytochemical contents, in vitro antioxidant and hypoglycaemic potentials of eleven indigenous plants using five different solvents. Putative hypoglycaemic agents from one of the most promising and readily available species were also identified using in silico molecular modelling. Secondary metabolites and their pharmacological activity have been reported as the basis for the wide use of plants in traditional medicine. However, due to the indiscriminate harvesting and environmental pressure, many valuable indigenous plant species have gone into extinction or are at least threatened. Moreover, plants' bioactive compounds are often produced in minute quantities, and prevailing environmental conditions further influence their concentrations in plants. Thus, due to indigenous plants' industrial and medicinal value, deliberate cultivation and elicitation strategies have been adopted for the en masse production of uniform indigenous plants and to influence the quality and quantity of their active principles. Thus, this study also assessed the effects of individual and co-inoculation of two isolated drought-resistant and growth promoting endophytes on the growth, drought tolerance, medicinal efficacy and metabolome changes in the leaves of Endostemon obtusifolius. In this research, the eleven plants were selected based on the traditional uses of the plants (or their related available species) for treating various ailments, including DM. The preliminary phytochemical quantification results revealed that the highest concentrations of phenolics, flavonoids and tannins were found in the crude extracts of Combretum krausssii, Lippia javanica, Psidium guajava, Pentanassia prenulloides, E. obtusifolius, Syzgium cordatum, Pachira aquatic and Catha edulis. The inhibitory effects of the crude extracts against the digestive enzymes α-amylase and α-glucosidase also showed that the crude extracts of C. edulis, C. krausssii, L. javanica, P. aquatica, P. guajava, P. prenulloides, E. obtusifolius and S. cordatum displayed excellent in vitro antioxidant and antidiabetic properties. These results validate the extensive use of these plants in the treatment of DM in many African communities. Furthermore, the 80% ethanol (v/v) leaf extract of S. cordatum (one of the most active and readily accessible specie from the previous study) was fractionated into four sub-extracts [petroleum ether (PE), dichloromethane (DCM), ethyl acetate (EtOAc) and water], and their phytochemical content, in vitro antioxidant and antidiabetic capacities were evaluated. Although the EtOAc extract was the richest of the sub-extracts in total phenolics, all four sub-extracts of S. cordatum showed good in vitro free radical scavenging and hypoglycaemic activities. In silico modelling evaluation of some (34) bioactive principles found in the Gas Chromatography-Mass Spectroscopy (GC-MS) analysis of the PE, DCM and EtOAc sub-extracts revealed that 21 compounds including andrographolide, benzylidene-iditol, cubenol and deoxyspergualin and bis[3,3,4,7-tetramethyl-1,3-2H-benzofuran-1-yl]-ether returned binding energy scores ≤ -7.5 kcal/mol against α-amylase and α-glucosidase enzymes indicative of their hypoglycaemic potentials. The physicochemical and toxicological properties of andrographolide, benzylidene-iditol, bis[3,3,4,7-tetramethyl-1,3-2H-benzofuran-1-yl]-ether and cubenol were predicted to be soluble with high gastrointestinal solubility and non-toxic following Lipinski's rule of five and Veber's rule. Thus, these results indicate that these compounds are potential candidates for oral drugs. The drought tolerance and in vitro plant growth-promoting properties of some endophytes isolated from E. obtusifolius (another active antidiabetic plant identified from the previous experiment) was evaluated. A total of 26 culturable endophytes (twelve fungi and fourteen bacteria) were isolated from the organs (leaf and root) of E. obtusifolius. These endophytic species displayed varying in vitro drought stress tolerance and plant-growth-promoting capacities. Two promising drought stress-tolerant and plant-growth-enhancing endophytic species (Fusarium oxysporum and Paenibacillus polymyxa) were subsequently identified using molecular tools. The identified bacterium (P. polymyxa) and fungus (F. oxysporum) exhibited a symbiotic relationship in an in vitro dual culture experiment. Paenibacillus polymyxa and F. oxysporum individual and co-inoculation differential effects on their host under varying water regimes was further evaluated. The plants were raised with or without endophyte infection under three watering regimes for two months, and their therapeutic efficacy, physiological, biochemical and metabolic responses were assessed. In this study, drought stress markedly affected the growth and hypoglycaemic potentials of E. obtusifolius. On the other hand, endophyte inoculation generally enhanced the dry shoot and root biomass, chlorophyll contents and fluorescence, total soluble sugar, relative water content, proline contents and superoxide dismutase activities in the leaves of E. obtusifolius, whereas their electrolyte leakage and malondialdehyde contents were lowered. As for phytochemical accumulation, while the total phenolic contents were slightly enhanced by the inoculation of endophytes in the leaves of E. obtusifolius, the flavonoid contents of the plant increased as the water deficit worsened. The EtOAc crude extracts' free radical scavenging capacity across the treatments remained unchanged; their in vitro α-glucosidase activity was negatively affected under moderate and severe drought stress but improved with endophyte inoculation. The metabolome difference between the twelve treatments was evaluated using GC-MS based metabolomics. The bi-plot PCA result revealed that the metabolome of fungal inoculated moderately stressed E. obtusifolius correlated less with the other E. obtusifolius plants under different treatments. Additionally, a heatmap of eight differential metabolites showed that the most responsive treatment (the co-inoculated severely drought-stressed plants) produced the highest quantities of non-protein amino acids and organic acids known to protect plant cells during abiotic stress. The leaf extracts of S. cordatum and E. obtusifolius showed remarkable antioxidant and antidiabetic potentials in this study. Although the putative active principles of these plants were identified using GC-MS analysis, proper isolation and quantification of these compounds can be explored by future studies. Moreover, some culturable endophytic species were isolated from the E. obtusifolius organs. Paenibacillus polymyxa and F. oxysporum showed their drought stress mitigating capacity in E. obtusifolius under varying water regimes. Although the concentration of some identified antidiabetic compounds in E. obtusifolius were up regulated, the mechanism involved in this observation requires further investigations. IQOQA Isifo sikashukela (Diabetes mellitus - DM) siwukungasebenzi kwezitho zangaphakathi okuhambisana nobuphezulu kwamazinga eglukhosi egazini, kuhambisana nokuphazamiseka kokugayeka kwamafutha, izakhamzimba ezingamaphrotheni nezinikimandla. IDM iyisifo esingelapheki, esingathelelani nesingelapheki ngokwemithi, esiphethe izigidi zabantu emhlabeni jikelele, kuphethe ngamazinga okugula aphezulu nokufa imbala, ikakhulu esimeni sokungakhawuki kwesifo ukhuvethe (coronavirus disease of 2019 - Covid-19). Ukusetshenziswa kwemithi eyizinqindimandla zesifo sikashukela yasentshonalanga sekudale izinselelo eziningi ngenxa yokucabangeka ukuthi kuthinta ukuphepha kwayo jikelele, ukwehluleka ukwelapha nezindleko. Iningi lemiphakathi yase-Afrika yethembele ezihlahleni eziyimithi (yokwelapha) nezingxube zayo ezinokuphilayo njengesizinda semithi ngokomphumela wesimo esingenele sezinsiza zezempilo, ukwesweleka kodokotela abelaphayo nokungameleki kwezindleko zokwelashwa. Ngalesi sizathu-ke, ingxenye yalolu cwaningo yahlola okuqukethwe ngamakhemikhali ezihlahla, ngezivikelizinhlayiya ezifakwe eshubhini lokuhlola nama-ejenti anezinga eliphezulu likashukela egazini avela kolunye uhlobo olwethembisayo nolutholakala kalula olwatholwa kusetshenziswa ukumodela kwemolekhula ngekhompiyutha. Kwabikwa ukuthi ukugayeka kokudla kwezinga lesibili, nomnyakazo wakho wezokwelapha ngemithi kuyisisekelo sokusetshenziswa kabanzi kwezihlahla emithini yendabuko. Nokho-ke, ngenxa yokuvunwa okunga okungakhethi nengcindezi yezokuvikelwa kwezemvelo, iningi lezinhlobo zezihlahla zendabuko zenani eliphezulu seziphelile kungenjalo-ke zisengcupheni. Naphezu-ke, izingxube ezinokuphilayo zivama ukukhiqizwa ngezamba ezincanyana, nezimo zokuvikelwa kwezemvelo ezikhona ziphinda zibe nomthelela wazo ogxile ezihlahleni. Ngakho-ke, ngenxa yenani lezihlahla zendabuko ngokwezimboni nakwezokwelapha, sekufakwe ngokwenhloso amasu okuzitshala nokuzivuselela ukuze zikhiqizele ngobuningi bazo nangokwefana kwezihlahla zendabuko nokuba nomthelela kohlonze nasebuningini kwemithetho esebenzayo. Ngalokho-ke lolu cwaningo lwahlola imithelela yokunye nokugonywa okuhlanganisiwe kokukhethiwe kokuhlala ezinhlayiyeni eziphilayo zokukhula, okumelana nesomiso nokukuthaza ukukhula, ukumelana nesomiso, ukwenza kahle ekwelapheni nezinguquko zokugayeka emaqabungeni e-Endostemon obtusifolius. Naphezu-ke, kwahlolwa i-ethanol (v/v) emuncwe emaqabungeni engama-80%, ye- S. cordatum (engenye yezinhlobo ezitholakala kalula nesebenzayo esukela ocwaningweni lwaphambilini) yacozululwa yaba ngokumunciwe okuncanyana [petroleum ether (PE), dichloromethane (DCM), ethyl acetate (EtOAc) namanzi], nokwamakhemikhali ezihlahla kwawo, nezivikelizinhlayiya ezifakwe eshubhini lokuhlola, nokukwazi ukulwa nesifo sikashukela. Nakuba isimuncwa i-EtOAc kwakuyiyo ecebe kunazo zonke izimuncwa ezincane zamafenolikhi aphelele, zonke izimucwa ezincane zozine ze- S. cordatum zakhombisa ukukwazi ukuthungatha okukhululekile nokunamandla nokulwa nezinga eliphezulu likashukela egazini. Ukuhlolwa kokumodelwa ngokwekhompiyutha, kwalokho okunokuphilayo okungama (34) ngokwemitheshwana yokunokuphilayo okwatholwa ohlaziyweni lwe-Gas Chromatography-Mass Spectroscopy (GC-MS) kwezimuncwa ezincane ze- PE, DCM ne-EtOAc zakhombisa ukuthi izingxube ezingama-21 kubalwa kuzo ne- andrographolide, benzylidene-iditol, cubenol ne-deoxyspergualin nebis[3,3,4,7-tetramethyl-1,3-2H-benzofuran-1-yl]-ether kwabuyisa imiphumela yamandla okubophezela ≤ -7.5 kcal/mol aphikisana nama-enzayimu e-α-amylase ne-a-glucosidase akhombisa ukuthi angakwazi ukulwa nezinga eliphezulu lesifo sikashukela. Okuqukethwe yiphysicochemical netoxicological andrographolide, benzylidene-iditol, bis[3,3,4,7-tetramethyl-1,3-2H-benzofuran-1-yl]-ether necubenol kwabikezelwa ngokukwazi ukugayeka ngokomgudusisu-mathumbu wokugayeka nokungabi nasihlungu kulandela umthetho kaLipinski wokuhlanu nomthetho weVeber. Ngalokho-ke, le miphumela ikhombisa ukuthi lezi zingxube zinganethuba lokungenela ukuba yimithi ephuzwayo. Kwahlolwa ukukwazi ukumela isomiso nokukwazi ukukhuthaza ukukhuliswa kwezinhlaka zezihlahla eshubhini lokho okuhlala kwezinye izinhlayiya zokuphilayo okwakhishwa ku- E. obtusifolius (esinye sezihlahla esilwa nesifo sikashukela esahlonzwa elingeni laphambilini). Isamba salokho okuhlala ezinhlayiyeni zokuphilayo sokukhulisekayo okungama-26 (ishuminambili lesikhunta namagciwane ayishumi nane) kwehluswa ezithweni (amaqabunga nezimpande) ze- E. obtusifolius. Lezi zinhlobo ezinokuhlala ezinhlayiyeni zalokho okuphilayo zakhombisa ukumelana nesomiso okwahlukene uma zifakwa eshubhini lokuhlola nokukwazi ukuba ngokukhuthazwa kokukhula kwezihlahla. Kwagcina sekutholwe izihlahla ezimbili ezikwazi ukumelana nokuhlukunyezwa yisomiso nokukwazi ukukhuthaza ukukhuliswa kwezihlahla kohlobo okuhlala kulo okuphilayo (Fusarium oxysporum nePaenibacillus polymyxa) kusetshenziswa amathuluzi anobumokhula. Ubugciwane obatholakala (P. polymyxa) nobukhunta (F. oxysporum) bakhombisa ubudlelwano obunokwencikana elingeni leshubhu lokuhlola elikhulisa ngakubili. Kwaphinda kwahlolwa iPaenibacillus polymyxa ne-F. oxysporum okungakunye nomgomo ohambisana nemithelela ehlusayo kokungumgcini wakho ezinhlotsheni zamanzi ezahlukene. Kwahlolwa izihlahla ezakhuliswa ngaphandle kokufakwa okuhlala kokuphilayo, ngaphansi kwezinhlobo ezintathu zokuchelela ngezinyanga ezimbili, nokusebenza kahle kokukwazi ukwelapha kwazo, ngokomzimba, ngokwamakhemikhali aphilayo nokwenza kwakho ekugayweni kokudla. Kulolu cwaningo, ukuhlukumeza kwesomiso kwaba nomthelela omkhulu ekukhuleni nokukwazi ukuvimba izinga eliphezulu likashukela kwe- E. obtusifolius. Ngakolunye uhlangothi, ukugonywa kwalokho okuhlala kokuphilayo kwavama ukukhuthaza ibhayomasi yezithombo nezimpande, okuqukethwe yiklorofili nokukhanyayo, ushukela oncibilika ngokuphelele, isikalo samanzi esinokuhambisana, okuqukethwe ngokwenziwa yiproline nesuperoxide okwenzeka ekuhlakazekeni kabili emaqabungeni e-E. obtusifolius, kanti kwehliswa ukuvuza kwe-elekthrolaythi yakho nokuqukethwe yimalondialdehyde. Ngokokunqwabelana kwamakhemikhali ezihlahla, ngenkathi okuqukethwe kwefenolikkhi kwakukhuthazwa ngukugonywa kwalokho okuhlala kokuphilayo emaqabungeni e-E. obtusifolius, okuqukethwe yiflavanoydi kwesihlahla kwakhula ngenkathi ukuncipha kwamanzi kudlanga. Okwe-EtOAc okungagayiwe okumuncwe ngomthamo okhululekile nokokuzingela okumawala kuzo zonke izinhlobo zokwelapha kwahlala kungaguqukile; ukusebenza kwe- α-glucosidase eshubhini lokuhlola kwatheleleka kabi ngaphansi kokuhlukunyezwa yisomiso esiphakathi nendawo nesadlulele, kodwa kwathuthuka ngokugonywa ngokuhlala kokuphilayo. Umehluko ekugayweni phakathi kokwelaphayo okuyishumi nambili wahlolwa kusetshenziswa i- GC-MS egxile kwezokugaya. Umphumela we bi-plot PCA waveza ukuthi ukugayeka kwesikhutha esigonyiwe kwahlukumeza ngokuphakathi kwendawo i- E. obtusifolius ehambisana kancane nezinye izihlahla ze- E. obtusifolius ngaphansi kokwelashwa okwahlukile. Ngokwengeziwe-ke, ibalazwe lokushisa kokugaya okunomehluko okuyisishiyagalombili kwakhombisa ukuthi ukwelapha okuzwela kakhulu (kwezihlahla ezigonywe ngokuhambisana ezihlukunyezwe kakhulu yisomiso) kwakhiqiza izibalo eziphezulu zezakhimaphrotheyni nama-esidi emvelo aziwa ngokuvikela izinhlayiya zezihlahla ngenkathi yokuhlukunyezwa ngokungenampilo. Okumuncwe emaqabungeni e- cordatum ne-E. obtusifolius kwakhombisa ubukhona kwezivikelizihlahla nokulwa nesifo sikashukela kulolu cwaningo. Nakuba imitheshwana esebenzayo ehlawumbiselwayo kwalezi zihlahla yabukwa kusetshenziswa uhlaziyo iGC-MS, ukwahluswa okufanele nokubalwa kwalezi zingxube kungahlolwa ezifundweni zangomuso. Naphezu-ke ezinye izinhlobo ezikhulisekayo zalokho okuhlala kokuphilayo zehluswa ezingxenyeni ze-E. obtusifolius. IPaenibacillus polymyxa ne-F. oxysporum kwakhombisa ukukwazi ukunciphisa ukukhandlwa yisomiso kwe- in E. obtusifolius okwalawulwa ngokuphakama, ukusebenza okwafakwa ekwethameleni kudinga okunye ukuhlolwa.