AMERICAN JOURNAL OF PHYSIOLOGY, BIOCHEMISTRY AND PHARMACOLOGY, 2018 VOL 7, NO. 1, PAGE 42–47 eJManager 10.5455/ajpbp.20180430035035

ORIGINAL RESEARCH Open Access Hypoglycemic effect of aqueous extract ofTelfairia occidentalis extract in alloxan induced diabetic wistar rats Okonkwo Chukwudi Onyeka.1, Egesie Gideon Umezurike.2, Maduka Stephen Ozoemena.3, Oguaka Victor Nwabunwanne.3 1Department of Human Physiology, Faculty of Basic Medical Sciences, College of Health Sciences, Nnamdi Azikiwe University, Nnewi Campus, 2Department of Human Physiology, Faculty of Basic Medical Sciences, College of Medicine, University of Jos, Jos, Nigeria 3Department of Human Biochemistry, Faculty of Basic Medical Sciences, College of Health Sciences, Nnamdi Azikiwe University, Nnewi Campus, Nigeria

ABSTRACT ARTICLE HISTORY Aim: This study was carried out to evaluate the effect of oral administration of aqueous Received 30 April 2018 extract of Telfaria occidentalis on insulin level of diabetic rats before and after 14 days Accepted 08 May 2018 of treatment. Published 12 June 2018 Methods: Diabetes was induced in the animals by an intraperitoneal injection of alloxan KEYWORDS monohydrate dissolved in sterile normal saline in a dose of 150 mg/kg body weight. After 72 hours of the injection, rats that are diabetic (indicted by hyperglycemia) were used for occidentalis; the experiment. Blood samples were collected from the tails of the rats and blood glu- diabetes mellitus; hyperglycemia; aqueous cose was determined using a glucometer. The rats were divided into Groups I–VI. Group extract I animals served as non-diabetic control and were administered 0.5 ml of water daily for 14 consecutive days. Groups II and III were non-diabetics that were administered daily with 150 mg/kg body weight and 300 mg/kg body weight of extracts (TO exts), respectively for 14 consecutive days. Group IV rats served as diabetic control and were administered with 0.5 ml of water daily for 14 consecutive days while groups V and VI were diabetic rats that were administered daily with 150 mg/kg body weight and 300 mg/kg body weight of TO exts, respectively for 14 consecutive days. Results: TO exts was shown to significantly (p < 0.05) lower the fasting blood glucose (FBG) in alloxan induced diabetic rats in a dose-related fashion, and also, showed signif- icant (p < 0.05) lowering of FBG in normal (non-diabetic) rats that received 300 mg/kg/ day of TO exts. Telfairia occidentalis showed no significant (p > 0.05) reduction in the FBG of normal (non-diabetics) rats that received 150 mg/kg/day of TO exts. Conclusion: Telfairia occidentalis might be producing its hypoglycemic effect via stim- ulation of insulin secretion from the beta cells of the islets of Langerhans or increased peripheral utilization of glucose by the cells.

Introduction Although various have been employed in Through the ages, plants have been used as sources traditional medicine in Nigeria to treat diabetes, a of drugs administered empirically or otherwise in the cure of diseases. Plants were also allegedly Telfairia occi- used to treat diabetes. Several plants have indeed dentalislot still remains is a tropical to be vinedone grown scientifically in West to Africa confirm as been investigated for their hypoglycemia proper- athe leaf efficacy vegetable of theseand for herbal its edible drugs. . It is also ties. Among these are 1) Telfairia occidentalis, 2) ocimum gratissimum, 3) mangifera indica, and 4) in Ibo Language in Eastern Part of Nigeria. Its scien- traditional Couboria vitata. known as fluted pumpkin, fluted gourd, or Ugu in

Contact Okonkwo Chukwudi O. [email protected] classification Department is of as Human follows: Physiology, Kingdom: Faculty Plantae, of Basic Medical Sciences, College of Health Sciences, Nnamdi Azikiwe University, Nnewi Campus, Nigeria.

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occidentalis is very rich in iron, antioxidants, phyto- chemicals (such as phenols), and ascorbic acid and TelfairiaDivision: Magnoliophyta,T. occidentalis Class:. Magnoliopsida, has been found to possess antimicrobial, free rad- Order:Telfairia , occidentalis Family: (), Cucurbitaceae, is Genus: culti- vated mainly, Species: in West Africa, especially in Nigeria, It has been reported that the aqueous extracts of Telfairiaical scavenging, occidentalis and therapeuticreduced blood activities glucose [21,22]. level (Telfairia occidentalis) is a creeping vegetative shrub and have hypoglycemia effects in glucose induced thatGhana, spreads and low Sierra across Leone the ground [1,2]. Fluted with large pumpkin lobed There is a considerable evidence to support the rolehyperglycemic of oxidative [23]. stress in the development of ath- sowing. and It is long grown twisting for its tendrilsleafy and [3]. its Harvestingoily . The of erosclerosis and related cardiovascular diseases. plantfluted is pumpkin a drought-tolerant; takes place dioeciously 120–150 days, perennial after The development of atherosclerosis involves lipids that is usually grown trellised. The young deposits on the inner walls of arteries, with subse- and leaves of the female are the main ingre- dients of a Nigerian soup, edikangikong. The large are composed mainly of cholesterol and cholesterol - esters,quent inflammationwhich are associated and scarring. with The foam fat cells deposits and tein, and can be eaten whole, ground into powder macrophages on the artery walls. Epidemiology for(up another to 5 cm), kind dark of soup,red seed or made is rich into in fata fermented and pro and experimental data show a clear correlation porridge. The fruit of the plant is large, weighing up between increased atherosclerosis and plasma to 13 kg, but inedible. cholesterol levels. Increased levels of low-density The leaves are cooked and eaten while the seeds lipoprotein are considered the principal casual factor. This becomes part of the atherosclerotic eaten, or ground into powder for soup. The seed canwhich also contain be fermented about 30% for severalprotein dayscan be and boiled eaten and as reducing cholesterol levels lowers the frequency ofprocess cardiovascular after oxidative disease modification.and improves Therefore,patholog- is being gradually investigated. Telrairia occiden- ical damage in heart failure. Although numerous talisslurry [4–7]. The medicinal importance of the plant trials of diet or drug-based cholesterol reduction - has provided compelling evidence that reducing is now known to possess anti-inflammatory cholesterol levels decrease the incidence of cardio- effect [8], anti-bacterial activity [9], erythropoi vascular disease, the therapeutic effects of Telfairia etic value [10], anticholesterolemicTelrairia occidentalis and immuneis a rich occidentalis, used in African medicines against sourcebuilding of properties minerals such [11], as and calcium, hypoglycemic phosphorous, effect pathological conditions caused by lipids disorders, [12–15]. The seed of remain uninvestigated. The plant is also rich in glycosides that yield cur- unsaturatediron, zinc, and fatty copper. acids which The seedoffer protective contains 47%role cubitacins and glucose on hydrolysis. The vitamin C againstoil. The atherosclerosis oil obtained from and thecardiovascular seed contains disease 61% matter, and Telfairia occidentalis has been reported tocontent protect of thisagainst plant cancer is about of 148.0the oesophagus, mg/100 g of oral dry [16]. The phospholipids, glycolipids,Telfairia andoccidentalis neutral - lipid contents of the seed are 58%, 26%, and 15%, ibility, and to improve circulation in the arteries of Thisrespectively research [17]. was The undertaken root of to screen the leaf smokers.cavity, and An stomach, extract from to maintain this plant blood has beenvessels shown flex ofhas Telfairia been screened occidentalis for hypoglycemiafor possible hypoglycemic activity [10]. to possess ant diabetic activity in both alloxan and - sess this activity. Leaves from this plant constitute anactivity, important since ingredientthe root has in beensoup makingconfirmed since to theypos Telfairiastreptozotocin occidentalis diabetic animals. However, there is are good sources of proteins, vitamins (B-complex), a dearth of information on the beneficial effect of minerals, fatty acids (linoleic and oleic acids), and with Telfairia occidentalis on the leaveslipid profile and theirof animals. wide- spreadBased acceptability, on the different this healthstudy benefitswas carried associated out to - investigate the hypoglycemic effects of Telfairia fibers. The seed contains 13% oil [18] and is used- occidentalis in both normal and diabetic rats. In blefor marmaladeis popularly manufacturing used in soup and [19] in and herbal cookie prepa for- the normoglycemic rats, the Beta cells of the pan- rationsmulations for [20].the management This darkish-green of many leafy diseases vegeta in creas which produce insulin are intact. But alloxan Nigeria. Studies have shown that the leaf of Telfairia destroys these Beta cells. Alloxan induced diabetic www.ajpbp.com 43 Okonkwo Chukwudi Onyeka., Egesie Gideon Umezurike., Maduka Stephen Ozoemena., Oguaka Victor Nwabunwanne. rats, therefore, no longer have functional pancre- atic Beta cells and have lost the capacity to secrete Telfairia occidentalis have been screened for hypo- insulin which is required for glucose absorption. It glycemic[17]. Both activity. the leaf This and research root (Unpublished was undertaken data) toof could be inferred, therefore, that the hypoglycemic screen the leaf of Telfairia occidentalis for possible effect of the seed extract is not mediated through hypoglycemic activity, since the seed has been con- the stimulation of insulin release from beta cells

Materialsfirmed to possess and Methodology this activity. likeOral the glucose sulphonylureas tolerance testing [24] but is a through standard other pro- Materials ceduremechanism used [25]. in the diagnosis of diabetes and in • 36 male Wistar rats, 14 weeks old, procured observed reduction of blood glucose concentration assessing extracts for hypoglycemic effect [26]. The • Telfairia occidentalis purchased from Terminus - from the University of Jos animal house. istered simultaneously) and an increase in blood in glucose loaded rates at 60 minutes (when admin • Table weighing scale Market, Jos. hour before glucose) shows that the seed extract is •  glucose level at 45 minutes (when administered 1 • Intragastric tubes not effective in reducing blood glucose concentra- Glucometer (on call plus Glucometer). tion in glucose loaded rats. This further buttresses •  •  the fact that the seed extract did not stimulate insu- Growers’ feeds procured from vital feeds, Jos. lin production by the pancreas. Alloxan monohydrate procured from Medicom It is therefore clear that the leaf extract may be •  DistilledLaboratory water Nig. Ltd Jos. useful in the management or ethnotherapy of type •  RotatoryMortar and Evaporator pestle 2 diabetes mellitus. The leaf of Telfairia occidenta- •  Normal saline lis •  • Airtight containers is known to contain about 14.5% carbohydrates. •  (Mostly sucrose, fructose, galactose, raffinose, and syringes) StandardSyringes rate (5 ml, cages. 2 ml, and insulin injection thestachyose), fatty acids 47% exhibiting lipids (phosphora high degree lipids—58%, of unsatu- •  rationglycol lipids—26%,contributed mainlyand neutral by the lipids—16%) C16 and C fattywith Methodology 18 Plant material contains alkaloids, steroids, tannins, and terpenes. acids [16]. And this study has shown that the leaf Telfairia occidentalis the constituents of the leaf of Telfairia occidentalis However, it is difficult at this stage to know which of was procured from Katako of the leaf extract Market, Jos. Identified by Professor Akueshi, plant didis/are not responsible reduce blood for glucosethe hypoglycemic −1level in the activity. diabetic It taxonomistThe air dried in the Telfairia Department occidentalis of Botany, was University pounded is also known that 250 mgdose. kg It may be that at that of Jos, Jos. - −1), the carbohydrates content tainers ready to use. rats like the 100 mg kg−1 and filtered. The powder was kept in airtight con glucosedose level level (250 thereby mg kg countering the hypoglycemic Extract of Telfairia occidentalis of the extract contributed significantly to the blood time that the leaf extract of Telfairia occidentalis Sixty gram of the dried powdered Telfairia occi- containseffect of thehypoglycemic extract. This constituents work shows which for couldthe first be dentalis was exhaustively extracted with water in a useful in the treatment of non-insulin dependent - (i.e., type 2) diabetes mellitus. centrated to dryness on a rotatory evaporator and soxlet extractor for 72 hours. The extract was con The medicinal importance of the plant is being weighed. The residual extract was dissolved in nor- gradually investigated. Telfairia occidentalis is now mal saline and used in the study. - Experimental induction of diabetes in rats cholesterolemicknown to possess and anti-inflammatory immune building effect properties [8], anti bacterial activity [9], erythropoietic value [10], anti- pholipids, glycolipids, and neutral lipid contents Male Wistar rats 14 weeks old (weighing between [11], and hypoglycemic effect [12–15]. The phos 130 and 200 g) bred in the animal house, Department of Human Physiology, Faculty of Medical Sciences, 44of the seed are 58%, 26%, and 15%, respectively A J Physiol Biochem Pharmacol • 2018 • Vol 7 • Issue 1 Hypoglycemic effect of aqueous extract of Telfairia occidentalis leaf

Table 1. Effect of oral administration of aqueous extract of Telfairia occidentalis on blood glucose level of non-diabetic rats & diabetic rats before and after 14 days of treatment (N = 6). Day 14 (Blood glucose in S/No Groups Day 0 (Blood glucose in mmol/l) mmol/l) i Non-diabetic control + 0.5 ml of water 79.17 ± 3.13 69.17 ± 3.68 ii Non-diabetic + 150 mg/kg TO ext 118.17 ± 7.15 a 87.33 ± 6.90 iii Non-diabetic + 300 mg/kg TO ext 52.83 ± 5.52 a 50.833 ± 1.35 iv Diabetic control + 0.5 ml of water 231.08 ± 29.66 388.67 ± 35.24 v Diabetic + 150 mg/kg TO ext 105.42 ± 10.12 77.83 ± 2.82 vi Diabetic + 300 mg/kg TO ext 55.33 ± 3.73 b 50.58 ± 2.26 Mean ± Standard Error of Means (SEM) for six determinations.

- After 14 consecutive days of treatment, blood mals were fed with grower feeds (procured from samples were collected from the rats for estimation University of Jos, were used in this study. The ani of their blood glucose levels. - drateVital Feed dissolved Jos) and in sterile allowed normal access saline to water in a freely. dose of Results The rats were injected with alloxan monohy Table 1 shows the effect of oral administration of - aqueous extract of Telfairia occidentalis on blood 150 mg/kg body weight intraperitoneally [27]. After 72 hours of the injection, rats with fast glucose level of non-diabetic rats & diabetic rats before and after 14 days of treatment. The effects collecteding blood from glucose the (FBG)tail of atthe or rats above and 126blood mg/dl. glu- cose(7.0 mmol/l)was determined were considered by glucose diabetic. oxidase Blood method was onof 14-daynormal oraland administrationalloxan induced of diabetic 0.5 ml ofrats water and and graded doses of TO ext (150 mg–300 mg/kg) of Trinder [28], using On Call Plus Glucometer Table 2 and 3 shows insulin levels after administra- ExperimentalMonitoring system design [29]. alloxan induced diabetic rats using immunoassay In the experiment, a total of 36 rats were used. analysis.tion of graded doses of TO ext on FBG in normal and starting the treatment. The rats were divided into to normal rats (group II) for 14 consecutive days sixDiabetes groups was after induced the induction in the rats of diabetes.72 hours Inbefore this The administration of 150 mg/kg/dayP of TO ext experiment, six rats were used in each group. did not cause any significant ( > 0.05) decrease Group I: ratsin their (group FBG. III)However, for 14 daily consecutive administration days causedof 300 water daily for 14 days. mg/kg/day ofp TO ext to the (non-diabetic) normal Group II Normal control ratsTelfairia given 0.5 occidenta ml/kg of- Also, daily oral administration of graded doses lis significant ( < 0.05) reduction in their FBG. : Normalweight) dailyrats given for 14 consecutive days. P Group III extract (TO ext) (150 mg/kg body reduction(150–300 inmg/kg) the blood of TO glucose ext to concentrationdiabetic rats groups when body weight) daily for 14 consecutive comparedV and VI caused with group significant IV. ( < 0.05) dose related : days. Normal rats given TO ext (300 mg/kg Group IV: - Discussion dailyUntreated for 14 diabetic consecutive rats days.(diabetic con Various in vivo models (e.g., diazoxide, alloxan, or Group V: trol rats) given 0.5 ml/kg of water streptozotocin-induced diabetic rats) are used in body weight) in saline solution daily evaluating medicinal plants with suspected hypo- Diabeticusing an ratsintragastric given TO tube ext (150for 14 mg/kg con- - secutive days. Group VI: Diabetic rats given Telfairia occidentia- glycemic potentials [29]. In this study, diabetes mel lis doselitus wasinduced induced diabetes using mellitusintraperitoneal in the treated injection rats of saline solution daily for 14 consecutive alloxan at a dose of 140 mg/kg of body weight. This- days. extract (300 mg/kg body weight) in betes mellitus by selectively destroying the pancre- atic72 hours beta cellsafter whichadministration. are involved Alloxan in the induces synthesis, dia www.ajpbp.com 45 Okonkwo Chukwudi Onyeka., Egesie Gideon Umezurike., Maduka Stephen Ozoemena., Oguaka Victor Nwabunwanne. storage, and release of insulin which is a peptide useful in the management of type II diabetes mel- hormone that regulate carbohydrate, protein, and - current administration of TO ext with conventional In this study, diabetes was fully induced as shown orallitus. hypoglycemic From the study, agents it has may been produce shown a synergisthat con- lipid metabolism [29].P tic effect that may be detrimental to the individual if not well monitored. It is therefore recommended ratsby the compared significant to (the < group 0.05) Ielevation rats. Oral in administra the fasting- that individuals placed on conventional oral hypo- blood glucose concentrations in the groups IV–VI glycemic agents should not consume Telfairis P - occidentalis supplement without consulting their eredtion ofthe TO blood ext(150 glucose mg–300 of diabetic mg/kg ratsof body in groups weight) V physicians. andfor 14 VI consecutive in a dose relateddays significantly fashion when ( < 0.05)compared low with group IV. References - consecutive days to normal (non-diabetic) rats Telfairia occidentalis. Also, administrationP of 300 mg/kg/day for 14 [1] EuphyticaAkoroda MA. Seed production and breeding poten tial of the fluted pumpkin, Telfaira 1990; 49(1):25–32. produced significant ( < 0.05) reduction in their occidentalis, West African vegetable crop. Econ Bot [2] Bosa EO, Mgbeogu EM. 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