JournalHypoglycemic of Multidisciplinary Activity of Cresentia Studies cujete L. O. J. T. Montenegro, B. A. Logramonte, Vol.(Calabash) 7,7, IssueIssue Fruit No.No. Pulp 2,2, pp.pp. in 25-38,25 Diabetic-38, DecemberDecember Male White 20182018 Mice J. M. S. Toledo, Z. S. Elumba, ISSN 2350-70202350-7020 (Print) & L. G. A. Samaniego ISSN 2362-94362362-9436 (Online) doi: http://dx.doi.org/10.7828/jmds.v7i2.1222 Hypoglycemic Activity of Cresentia cujete L. (Calabash) Fruit Pulp in Diabetic Male White Mice Lorelie Gloria A. Samaniego, Ofel Jireh T. Montenegro, Benjo A. Logramonte, Joliesa Mae S. Toledo, Zeus S. Elumba Department of Biology, Central Mindanao University, Musuan, Maramag, Bukidnon Corresponding author: Zeus S. Elumba, email: [email protected] Abstract Cresentia cujete L. has been postulated to lower blood sugar level yet current research is insufficient to prove this claim. Hence, this study was conducted to investigate the hypoglycemic activity of C. cujete L. fruit pulp decoction and fresh pulp extract in experimental diabetic male white mice. The behavior of the mice was also observed. Twelve male diabetic white mice were divided - + into four groups (T0 = distilled water, T0 = metformin, T1= Cresentia cujete L. fruit pulp decoction, T2= Cresentia cujete L. fruit pulp fresh extract) and subjected to treatment by gavage for 28 days. Fasting blood sugar (FBS) level was determined before diabetes induction, two days after diabetes induction (the onset of diabetes or Day 0), on Day 15 and Day 29. Data analysis employed Analysis of Variance in Complete Randomized Design and Tukey’s HSD (honest significant difference) Test. At Day 0, all mice had high FBS (>140 mg/dl). At Day 15, although ANOVA showed a highly significant difference, the effect of T1 and T2 was similar. At Day 29, the FBS level in the four groups varied significantly. Symptoms of diabetes (polydipsia, polyphagia, body weakness) were observed at the start of the study. During the study, the mice had reduced food intake and became more active. Results suggest that fresh fruit pulp extract is more efficient in lowering blood glucose than the decoction. Keywords: decoction, diabetes, extract, polydipsia, polyphagia 25 25 JournalHypoglycemic of Multidisciplinary Activity of Cresentia Studies cujete L. O. J. T. Montenegro, B. A. Logramonte, Vol.(Calabash) 7, Issue Fruit No. Pulp2, pp. in 25-38, Diabetic December Male White 2018 Mice J. M. S. Toledo, Z. S. Elumba, ISSN 2350-7020 (Print) & L. G. A. Samaniego ISSN 2362-9436 (Online) doi: http://dx.doi.org/10.7828/jmds.v7i2.1222 Introduction Diabetes mellitus is a major human health concern today and is considered as one of the leading causes of death worldwide (Vats et al., 2004; Kumar & Loganathan, 2010; Rohilla & Ali, 2012). It causes alterations in carbohydrate and lipid metabolism and complications like end-stage renal disease (ESRD), heart attack, stroke, and nerve disorders such as retinopathy and lower extremity amputations (Firdous et al., 2009; Mohini et al., 2012; Kasper et al., 2015). There are currently available therapeutic options for diabetes like oral hypoglycemic agents and insulin but many herbal alternatives have been recommended for the control of diabetes. In fact, there is a continuing effort to find substitutes from plant sources for this disease. Cresentia cujete L., commonly known as calabash, is popular in the traditional system of medicine. The leaves and bark showed antimicrobial activity (Binutu & Lajubutu, 1994; Mahbub et al., 2011) and are used as anthelminthic laxative and for treating skin diseases (Stuart, 2012). The leaves have anti-inflammatory and anti-diabetic properties (Pöll, 1997; Lim, 2012) while the fruit can be used to treat colds, bronchitis, cough, asthma, urethritis, and various digestive disorders such as diarrhea and stomachache (Wang et al., 2010). The fruit is edible and the pulp contains phytochemicals such as alkaloids, flavonoids, saponins, tannins, and phenols; minerals like sodium, potassium, calcium, phosphorus, and magnesium; and vitamins such as A, C, E, B1 (thiamin), B2 (riboflavin), and niacin (Bever, 1980; Ogbuagu, 2008; Ejelonu et al., 2011). Some people postulated that this plant can help lower blood sugar yet current research is insufficient to conclude this pharmacological claim. Thus, this study was conducted in order to investigate the hypoglycemic activity of C. cujete L. fruit pulp in alloxan-induced diabetic male white mice. Specifically, it aimed 26 26 HypoglycemicJournal of Multidisciplinary Activity of Cresentia Studies cujete L. O. J. T. Montenegro, B. A. Logramonte, (Calabash)Vol. 7, Issue Fruit No. Pulp 2, pp. in 25 Diabetic-38, December Male White 2018 Mice J. M. S. Toledo, Z. S. Elumba, ISSN 2350-7020 (Print) & L. G. A. Samaniego ISSN 2362-9436 (Online) doi: http://dx.doi.org/10.7828/jmds.v7i2.1222 to compare the efficacy of fruit pulp decoction and fresh pulp extract in lowering the blood glucose of experimental mice with the hypothesis that there is no significant change in the blood glucose of diabetic male white mice after treatment with C. cujete L. fruit pulp. The behavior of the mice throughout the study was noted. The result of this study can validate the claim that C. cujete L. can be used as an alternative medicine for the management of diabetes mellitus. The study did not include analysis of other blood chemistry due to capability constraints. Determination and quantitation of phytochemicals in the extracts were also excluded. Pathological and toxicity effects of the extracts on the different organs of mice and the mechanism of action of the phytochemicals were not studied as well. Only male white mice were used because female mice have hormonal fluctuation during the estrous cycle which may influence the results of the experiment (Marcondes et al., 2002). Materials and Methods Twelve alloxan-induced diabetic male white mice were individually caged and divided into four treatment groups (3 mice in each group) adopting the Complete Randomized Design (CRD). Diabetes was induced by single intraperitoneal injection of alloxan monohydrate. The negative control group (T0ˉ) was given 0.30 ml + distilled water while the positive control (T0 ) was given 10.0 mg of Metformin HCl dissolved in 0.30 ml distilled water. Treatments 1 (T1) and 2 (T2) were given 0.30 ml C. cujete L. fruit pulp decoction and fresh pulp extract, respectively. Treatments were administered at 6:00 in the morning for 28 days using improvised gavage. The fruit pulp decoction was prepared by slow boiling of 100.0 g minced C. cujete L. in 400.0 ml water for an hour. Bubbles were scooped out and were discarded. The decoction was allowed to cool down, filtered using fine 27 27 JournalHypoglycemic of Multidisciplinary Activity of Cresentia Studies cujete L. O. J. T. Montenegro, B. A. Logramonte, Vol.(Calabash) 7, Issue Fruit No. Pulp2, pp. in 25-38, Diabetic December Male White 2018 Mice J. M. S. Toledo, Z. S. Elumba, ISSN 2350-7020 (Print) & L. G. A. Samaniego ISSN 2362-9436 (Online) doi: http://dx.doi.org/10.7828/jmds.v7i2.1222 mesh cloth and filter paper, and refrigerated until use. Fresh pulp extract was prepared using a juicer. Individual weight of the mice was determined before treatment administration (Day 0) and at the end of the experimental period (Day 29) using a weighing balance. The behavior of each mouse was observed every morning and afternoon. Blood was drawn from the tail of each mouse. Blood glucose level was measured four times during the study: before diabetes induction, two days after the induction (Day 0), Day 15, and Day 29. The BETACHEK G5™ glucometer was used. Blood glucose level over 140.0 mg/dl is considered diabetic (Ahmed et al., 2010). One-Way Analysis of Variance (ANOVA) and Tukey’s (HSD) test were employed to analyze the data. Results and Discussion Fasting Blood Glucose Level (FBGL) Two days after administration of alloxan monohydrate (Day 0), the blood glucose level of the experimental mice reached the diabetic level (>140 mg/dl) (Table 1). This condition is partly due to partial necrosis of the pancreatic beta-cells (Lenzen, 2008) resulting in low insulin secretion and an increase in blood glucose level. As shown in the results, both fruit pulp decoction and fresh pulp extract were able to significantly lower (p<0.05) the blood glucose levels of the diabetic mice at days 15 and 29, although the result for T1 was still not within the normal value. Possibly, C. cujete L. fruit, like other hypoglycemic plants, stimulated the residual pancreatic beta-cells to produce more insulin (Chude et al., 2001) and stimulated regeneration of the beta-cells (Abdel-Rahim et al., 2011). 28 28 HypoglycemicJournal of Multidisciplinary Activity of Cresentia Studies cujete L. O. J. T. Montenegro, B. A. Logramonte, (Calabash)Vol. 7, Issue Fruit No. Pulp 2, pp. in 25 Diabetic-38, December Male White 2018 Mice J. M. S. Toledo, Z. S. Elumba, ISSN 2350-7020 (Print) & L. G. A. Samaniego ISSN 2362-9436 (Online) doi: http://dx.doi.org/10.7828/jmds.v7i2.1222 Table 1. Mean fasting blood glucose level (mg/dl) of the experimental mice. Treatment Day % Difference Groups 0 15 29 Reduction - a a To 208.7±14.19 217.0±12.72 223.0±12.73 14.3 (6.41) + c d To 178.3±36.50 140.0±8.54 92.3±7.09 -86.0 93.17 b b T1 182.7±26.83 176.0±26.91 158.3±18.77 -24.4 16.15 b c T2 188.0±3.60 177.3±4.51 137.7±4.16 -50.3 36.53 Means with the same letter are not significantly different at P = 0.05 using Tukey’s (HSD) Test. Bever (1980), Ogbuagu (2008) and Ejelonu et al.