IJBPAS, July, 2018, 7(7): 1374-1383 ISSN: 2277–4998
Dillenia philippinensis, AN ILONGOT ETHNOBOTANICAL OF AURORA, PHILIPPINES SHOWS ANTIDIABETIC ACTIVITY
MARY GRACE B. ANGELES, CYNTHIA C. DIVINA AND KHRISTINA G. JUDAN CRUZ* Department of Biological Sciences, Central Luzon State University, Science City of Muῆoz, Nueva Ecija, Philippines *Corresponding author: Khristina G. Judan Cruz E-mail: [email protected] th st th st Received 28 Feb. 2018; Revised 31 March. 2018; Accepted 24 April 2018; Available online 1 July 2018
https://doi.org/10.31032/IJBPAS/2018/7.7.4494 ABSTRACT Three plants with claims of antidiabetic potential: Dillenia philippinensis (katmon bark), Codiaeum variegatum (San Francisco) and Hydrocotyle vulgaris (Gutokola) were tested for their antidiabetic potential on lowering the blood glucose level in alloxan-induced diabetic mice. D. philippinensis (Katmon Bark) effectively lowered the blood glucose levels of di- abetic mice. Upon oral administration of the extract D. philippinensis, the blood glucose level had lowered by 15.92% at 30 mins, 5.57% at 120 mins and 12.43% 24 hours post treatment administration with significantly comparable values on the positive control (glibenclamide) with percentage mean values lowered by 4.88% at 30 mins, 15.5% at 120 mins and 5.8% at 24 hours respectively. Keywords: alloxan-induced diabetic mice, blood glucose level, Dillenia philippinensis
INTRODUCTION are now intensifying efforts on alternative Management of diabetes continues and complementary medicines to proffer to be a challenge to the medical communi- lasting solution or at least control some ty [1]. Studies have shown that due to the harmful effects of synthetic drugs[2]. inherent side effects of common oral hy- Choosing specific antihyperglycemic agent poglycemic synthetic drugs, researchers is predicated on their effectiveness in lo-
1374 IJBPAS, July, 2018, 7(7) Khristina G. Judan Cruz et al Research Article wering blood glucose, extra glycemic ef- eral studies were done, however, studies fects that may reduce long-term complica- on the ethnobotanicals from the Ilongot- tions, as well as issues on safety, tolerabil- Eǵongot community of Bayanihan, Maria ity, and expense [3]. Aurora, Aurora are still limited. This study generally highlights the antidiabetic poten- The Philippines holds a high num- tial of Dillenia philippinensis. ber of medicinal plant species entailing a broader scope for healing. However, with MATERIALS AND METHODS further economic development, there is Collection of Plant Samples rapid disappearance of traditional culture Three ethnobotanical plants previously and natural resource[4]. Various plants surveyed [9] with the permission of elders have been traditionally used throughout and barangay officials were used in the in history to reduce blood glucose and im- vivo testing for its antidiabetic potential, prove diabetes and its complications de- these were: Codieum variegatum leaves, spite insufficient scientific information[5]. Dillenia philippinensis bark and Hydroco- Although several herbal drugs, in spite tyle vulgaris leaves. Plant samples were their pronounced folkloric activity and collected in Bayanihan, Maria Aurora, Au- known for having anti-diabetic property rora. and their therapeutic evidence within the traditional healthcare system are reliable, Extraction Procedure most have not been formulated as drugs The collected plant samples were [6]. washed using distilled water and placed separately into clean plastic bags. These Ethnobotanical information indi- samples were air-dried for 5-7 days at cates that more than 1,200 species of room temperature until it was pulverized plants are identified as having hypogly- and stored in a resealable plastic bag. The cemic activity [7]. These medicinal plants pulverized plant samples weighing 20 are used to treat hypoglycemic or hyper- grams were extracted in 600 ml distilled glycemic conditions that are of considera- water in 80-90°C water for two hours. The ble interest for ethnobotanical community extracted plant samples were lyophilized as they are recognized to contain valuable for 72 hours and refrigerated. medicinal properties with large numbers showing varying degrees of hypoglycemic and anti-hyperglycemic activity [8]. Sev-
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Preparation and Handling of Test Mice The treatments were taken orally with Tests were done on animals fol- the aid of an improvised gavage according lowing the Institutional Animal Care and to their treatment designation. Two con- Use Committee (IACUC) policies, proce- trols were used: the negative diabetic con- dures and guidelines [10]. Male mice trol treated with physiological saline, and weighing 20-22 grams male mice aged the positive diabetic control treated with eight weeks or older were used and pro- standard antidiabetic drug Glibenclamide vided with ad libitum standard mice pellet at an oral dose of 3mg/kg body weight. diet and distilled water. Experimental ani- Three groups constituted the mice treated mals were conditioned and housed in an with designated lyophilized ethnobotanical air-conditioned room with normal 12 extracts with a selected dose of 200 mg/kg hours day and night cycle. The mice were body weight dissolved in a 1% concentra- divided into five groups containing five tion. All mice were treated with a 0.1 ml mice each in improvised polypropylene dosage with saline solution as the diluent cages. for the administration [14] [15] [16].
Induction of Diabetes to Experimental Glucose Monitoring of Blood Mice Blood samples were withdrawn Diabetes was induced via intraperito- from the animals by gently cutting a small neal injection of alloxan-monohydrate portion of the tail tip of the diabetic mice. with 150 mg/kg b.w. dosage [11]. Induc- The tips of the tail were sterilized by tion was made to overnight fasted animals swabbing with 70% ethanol after the oper- (8-12 hours fasting period) with access to ation. Test mice were bled according to water[12]. After the induction, animals the selected time frame of 0 hour or the were given 5% sucrose solution to over- baseline blood glucose level value and for come the induced hypoglycemia[13]. After every 30 mins, 120 mins, and 24 hours 72 hours, diabetic state were verified and upon treatment. Blood glucose levels were mice with blood glucose level above 250 recorded in mg/dl. mg/dl were declared as diabetic and used in the study. RESULTS AND DISCUSSION The results revealed that the blood Oral Induction and Treatment glucose level (mg/dl) among treatment Administration means recorded at 30 mins, 120 mins and
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24 hours after treatment administration 15.92% at 30 mins, 5.57% at 120 mins and showed to have significant differences 12.43% after 24 hours observation period. among groups at <0.05 level of significant The positive control glibenclamide had (Table 1). The mean change in the blood percentage mean values lowered by 4.88% glucose level recorded after 30 mins re- at 30 mins, 15.5% at 120 mins and 5.8% at vealed that the positive control (gliben- 24 hours respectively. The percentage clamide) and the extract of D. philippinen- blood glucose level of C. variegatum and sis were statistically comparable on lower- H. vulgaris consistently increased from ing the blood glucose level in diabetic initial value up to the 24 hours period of mice. The negative control (saline), as well observation. Despite the decrease in blood as the treatment consisting the extracts of glucose levels, all treatment groups failed C. variegatum and H. vulgarisare signifi- to reach the normal blood glucose level. cantly different to the positive control at On a separate treatment set up constituting the same observation period. of normal mice, comparison revealed that Furthermore, at 120 mins and 24 diabetic group failed to reach the normal hours observation period, it can be noticed blood glucose level range of 120-123 that the extract D. philippinensis still ap- mg/dl recorded in normal mice up to 24 peared to be statistically equal with the hours treatment period. mean change in the blood glucose level of Phytochemicals known to be con- the positive control (glibenclamide) having tained in D. philippinensis such as alkalo- its consistent effectivity on the lowering of ids, saponins, phenols, flavonoids and tan- the blood glucose level. nins [17] are known to decrease blood glu- Figure 1 shows the mean percen- cose aside from the additional anti-diabetic tage change in the blood glucose level rec- action of mineral and trace elements orded from the initial or the baseline blood present such as zinc (Zn), Chromium (Cr), glucose value and after the treatment ad- manganese (Mn), selenium (Se), Potas- ministration recorded at 30 mins, 120 mins sium (K), Vanadium (V) and Calcium and 24 hours. The graph depicts the per- [18]. D. philippinensisis reported to pos- centage change in the blood glucose level sess several bioactive compounds which recorded from the initial value up to the 24 are of potential use against diabetes [19]. hours observation period. Upon oral ad- Although there are no reported studies on ministration of the extract D. philippinen- the antidiabetic potential of the bark of D. sis, the blood glucose level lowered by philippinensis, research on the ethanol leaf
1377 IJBPAS, July, 2018, 7(7) Khristina G. Judan Cruz et al Research Article extracts indicate its hypoglycemic activity flavonoids [25] [26] [27] which are report- due to the presence of compounds such as edly produced by C. variegatum may not tannins and flavonoids [20]. Flavonoids have been extracted through the water ex- such as polyhydroxylated flavonol and traction method. Extraction of phytochem- myricetin have insulin omimetic properties icals such as flavonol, terpenoids and alka- that stimulate lipogenesis and transport of loids [28] may be improved by using glucose in the adipocytes, hence, have po- another extraction solvent such as ethanol. tential on decreasing the blood sugar levels Aqueous extraction methods, although [21]. On the other hand, all forms of tan- shown to have similar efficiency as etha- nins, in clinical terms, may participate in nol [29], may not have been as efficient in the management of blood sugar level. This extracting the compounds present in C. is demonstrated to stimulate the receptor variegatum, hence, showing negative re- cells to utilize carbohydrates [22] and are sults . In addition, alkaloids are important also described as polyphenolic compounds compounds due to its blood sugar lowering commonly found in both gymnosperms effect [30] but these can only be extracted and angiosperms [23] hence, usually occur through the ethanol extraction method. in almost all parts of a plant species, par- Phytochemicals in H. vulgaris such as sa- ticularly in the bark, leaves, fruits and ponins [31] have been demonstrated to de- roots[24] and can be extracted through crease blood sugar levels in elderly diabet- aqueous extraction. The possible presence ic patients [32] and reported to have anti- of tannins in D. philippinensis bark may diabetic effect [17]. However, this study have contributed to its antidiabetic poten- revealed that H. vulgaris has no significant tial. As such, D. philippinensis extract effect on the lowering of blood glucose should be evaluated to further confirm its level on diabetic mice. For some reason, active components and its bioactivities. saponins aside from possessing anti- microbial activity, its toxic effects may The mode of plant extraction may explain why this compound do not possess have also affected the effectivity of the medicinal significance [33]. Also in some plants on the antidiabetic potential testing. cases, because saponins possess carbohy- To mention, compounds with possible an- drate moieties, this compound is capable tidiabetic properties such as valuable sec- of promoting further increase in the blood ondary metabolites alkaloids, terpenes and glucose level [34].
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Table 1: Mean change in blood glucose level (mg/dl) recorded at 30 mins, 120 mins and 24 hours upon treatment administration Treatments After 30 After 120 mins After 24 hrs mins
Negative Control -3.00b -14.00b -15.00b
Positive Control 28.00a 89.00a 33.33a
D. philippinensis 77.67a 27.67ab 60.67a
C. variegatum -28.33b -20.33b -40.33b
H. vulgaris -52.00b -72.00b -64.33b
*Means with similar letters per columns are not significantly different at 5% level of significance; Negative value indicates an in- crease in blood glucose level; Positive value indicates effectivity on the lowering of blood glucose level.
150.
120.
90.
60. Diabetic control Glibenclamide 30. D. philippinensis C. variegatum % % glucosein Changelevel blood H. vulgaris 0. 0 hour 30 mins. 120 mins. 24 hrs.
Figure 1: Mean percentage change of blood glucose levels (mg/dl). CONCLUSION Animal Medical Center, Central Luzon Sta- D. philippinensis bark lyophilized te University, Science City of Munoz, Nu- plant extract possesses antidiabetic poten- eva Ecija, Philippines for the use of their tial. The extract shows consistent effectivi- laboratories. ty in lowering the blood glucose level of REFERENCES the diabetic mice up to the 24 hours. [1] PATIL, A., JADHAV, V., ACKNOWLEDGEMENT ARVINDEKAR, A. & MORE, T. The authors acknowledge the per- (2014). Antidiabetic Activity of mission, assistance and support of the Ilon- Maesa indica (Roxb.) Stem Bark in got-Eǵongot community of Bayanihan, Streptozotocin Induced Diabetic Rats. Maria Aurora, Aurora, Philippines. This American Journal of Phytomedicine piece of work is dedicated to them. The au- and Clinical Therapeutics, 4, 14-16. thors acknowledge the support of the Co- [2] KANE, M.P., ABU-BAKER, A. & llege of Veterinary Science and Medicine- BUSCH, R.S. (2005). The Utility of
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