EVALUATION OF ANTIDIABETIC POTENTIAL OF KANDELIA CANDEL AND RHIZOPHORA APICULATA- AN IN VITRO APPROACH

EVALUATION OF ANTIDIABETIC POTENTIAL OF KANDELIA CANDEL AND RHIZOPHORA APICULATA- AN IN VITRO APPROACH

Arun Kashivishwanath Shettar and Ankala Basappa Vedamurthy ^*

PG Department of Studies in Biotechnology and Microbiology, Karnatak University, Dharwad - 580003, Karnataka, India.

ABSTRACT: Objective of the present work was to evaluating the antidiabetic property of Kandelia candel and Rhizophora apiculata extracts by using in vitro assays. The serial exhaustive extraction was carried out with a series of solvents:  chloroform, ethyl acetate, methanol, ethanol and water with increasing polarity using Soxhlet apparatus. The concentrated and dried extracts were evaluated for antidiabetic activity by employing standard in vitro techniques (Alpha-amylase and glucose uptake assay using yeast model). In vitro antidiabetic studies show that in case of Kandelia candel methanol extract showed higher antidiabetic activity where as incase of Rhizophora apiculata aqueous extract exhibited significant activity when compared to other solvent extracts.  Results confirm that aqueous extract of Rhizophora apiculata exhibited highest antidiabetic activity among all extracts. This study provides scientific evidence that the leaves of Kandelia candel and Rhizophora apiculata have anti-diabetic efficacy. However further comprehensive chemical and pharmacological investigation should be carried out to isolate the active compounds and appropriate elucidation of its mechanism of action and it helps in the development of new pharmaceuticals to treat Diabetes mellitus.

Kandelia candel, Rhizophora apiculata, Alpha-amylase assay, Glucose uptake assay, In vitro Antidiabetic activity

INTRODUCTION: Diabetes mellitus (DM) is a non-communicable disease often genetic in nature but can be developed due to life style and habits ^{1, 2}. DM is a set of metabolic disorder with a common feature of high blood glucose level. It is characterized by increased blood glucose, triglyceride and hypo insulinemia that may lead to decrease in both insulin action and secretion ^{3, 4}. Diabetes mellitus ranks highly with the top ten disorders which cause mortality throughout the world and is affecting approximately 30% of the worldwide population ^5-7.

Uncontrolled Diabetes mellitus is often associated with many complications which include development of micro and macro vascular complications such as neuropathy, nephropathy, retinopathy and cardiovascular diseases ^{8, 9}. Normally blood glucose levels are controlled by insulin produced by pancreas ¹⁰. Treatment of diabetes mainly focuses on reducing rise in blood sugar and subsequent complications. Inhibitors of α-amylase are currently used as oral hypoglycemic agents for treatment and management Diabetes.

Currently available anti-diabetic drugs possess severe side effects such as risk of hypoglycemia, anemia, and cholestatic jaundice ¹¹. Since from immemorial time traditional medicines have been used to treat Diabetes mellitus ¹². Many natural medicines have been reported to show anti-diabetic activity through different mechanisms.

Plants have been reported to reduce the absorption of glucose by retarding the digestive enzymes i.e. α-amylase and α-glucosidase from pancreas and gut which in turn helps preventing the high rise of glucose in blood ^13-16. Several works demonstrated that many medicinal plants can be used to treat and management of diabetes and already many medicinal plants have been reported for potent antidiabetic activity ^17-23.

In the present study Kandelia candel and Rhizophora apiculata plants were selected for screening antidiabetic property. Kandelia candel is the mangrove tree belonging to Rhizophoraceae family which is distributed along the western region of India. K.candel whole plant is reported to have antidiabetic activity ^{24, 25}. In fact Rhizophoraceae species are known to have pharmacological activities. Methanolic extract of K.candel is used as antihyperglycemic agent in India ²⁶ and bark, flowers and leaves were reported to have antiviral and antimicrobial properties ^{27, 28}. Rhizophora apiculata is the tree species of mangrove tree belonging to rhizophoraceae family.

In Malaysia, the leaves of R. apiculata are assayed as antibreast cancer ²⁹. Studies on HPLC investigation of R. apiculata have shown the presence of catechin monomer, an antioxidant flavonoid ³⁰. This plant is reported to possess anti-inflammatory and anti tumor properties and is also used to regulate the antioxidant enzymes in biological system ³¹. Presence of tannins is reported in the bark of R. apiculata, which is known to possess antibacterial and antiviral properties ^32-34. Bark of R. apiculata is used as a traditional medicine in the treatment of diarrhoea and wounds ^{35, 36}. In Malaysia, pyroligneous acid from R. apiculata species have been used as sterilizing agent, deodorizer, fertilizer, antimicrobial agent and growth promoting agent ³⁷.

Alkaline extract from leaf of R. apiculata reported to inhibit the HIV replication and HIV induced cytopathic effects. Some other studies have confirmed the antiviral property of R. apiculata extracts, which may be due to presence of antipolysaccharide in the extracts that acts as an antiviral agent ³⁸. With this background, the present study was undertaken to evaluate antidiabetic properties of Kandelia candel and Rhizophora apiculata plants collected from Mangrove region of Karnataka, India by using in-vitro assays.

MATERIALS AND METHODS:

Plant collection: Leaves of Kandelia candel and Rhizophora apiculata were collected from Mangrove region, Sadashivghad, Karwar, Uttar Kannada District, Karnataka, India during the period of May, 2015. The leaves were identified and authenticated by Dr. Kotresha K, Dept of Botany, Karnatak Science College, Dharwad; Karnataka by referring to the voucher specimen deposited in the Dept of Botany, Karnatak Science College, Dharwad, Karnataka (Voucher specimen No 003 and 004). Fresh plant leaves material was collected and washed under running tap water, shade dried and then homogenized to coarsely powder. The powder was stored in airtight containers at -20°C for further use for crude solvent extraction.

Crude Extraction: Coarsely powdered dried leaves of Kandelia candel and Rhizophora apiculata [100g each] were subjected to successive solvent extraction using soxhlet apparatus separately. The extraction of each plant leaves material was done with different solvents in their increasing order of polarity which includes chloroform, ethyl acetate, methanol, ethanol and distilled water. Each time the plant material was dried and later extracted with next high polar solvents (following the strategy of extraction in series of increasing the solvent polarity). All extracts were concentrated in Buchi rotary evaporator, followed by removal of traces of solvent by using desiccator. 

Evaluation of Antidiabetic activity by using in vitro assays:

Alpha -amylase inhibitory assay: The Alpha-amylase inhibitory assay for different solvent extracts of Kandelia candel and Rhizophora apiculata were evaluated according to a previously described method by Malik and Singh et al., (1980) with slight modification ³⁹. In brief, 0.5 ml of extract was mixed with 0.5 ml of α-amylase solution (0.5 mg/ml) with 0.02 M sodium phosphate buffer (pH 6.9 with 0.006 M NaCl).

The mixture was incubated at room temperature for 10 min and 0.5 ml of starch solution (1%) in 0.02 M sodium phosphate buffer (pH 6.9 with 0.006 M NaCl) was added. The resulting mixture was incubated at room temperature for 10 min, and the reaction was terminated using 1mL of dinitrosalicylic acid color reagent. At this time, the test tubes were placed in a water bath (100 ⁰C and 5 min) and cooled until room temperature was reached. The mixture was then diluted with 10 ml of deionized water, and absorbance was determined at 540 nm. The absorbance of blank (buffer instead of extract and amylase solution) and control (buffer instead of extract) samples were also determined. Acarbose was used as standard drug. The inhibition of α-amylase was calculated using the following equation:

% inhibition of α-Amylase = (Abs _control – Abs _sample) / (Abs _control) × 100

Where Abs _control corresponds to the absorbance of the solution without extract (buffer instead of extract) and with α -amylase solution and Abs _sample corresponds to the solution with extract and α -amylase solution.

Glucose uptake in Yeast cells: Glucose uptake assay by using yeast cells was performed according to the method of Cirillo et al., (1963) ⁴⁰. The commercial baker’s yeast in distilled water was subjected to repeated centrifugation (3,000×g, 5 min) until clear supernatant fluids were obtained and 10% (v/v) of the suspension was prepared in distilled water. Various concentrations of solvents extract of Kandelia candel and Rhizophora apiculata (50 to 250 μg/mL) were added to 1mL of glucose solution (5 mM) and incubated together for 10 min at 37 ⁰C. Reaction was started by adding 100 μL of yeast suspension followed by vortexing and further incubation at 37 ⁰C for 60 min. After 60 min, the tubes were centrifuged (2,500 × g, 5 min) and amount of glucose was estimated in the supernatant. Metronidazole was used as standard drug. The percentage increase in glucose uptake by yeast cells was calculated using the following formula:

Where, Abs sample is the absorbance of test sample and Abs control is the absorbance of control reaction (containing all reagents except the test sample). All the experiments were carried out in triplicates.

Statistical analysis: All experiments were performed in triplicates (n=3) and the data are presented as the mean ± standard error. Differences between the means of the individual groups were analyzed using the analysis of variance procedure of SPSS software 20 Version (IBM). The significance of differences was defined at the P < 0.05 and P < 0.01 level.

RESULTS AND DISCUSSION: Diabetes mellitus is one of the three known major killers that cause deterioration in human health after cancer and cardio-cerebral vascular diseases and it is rising over the years ⁴¹. The increasing prevalence of diabetes mellitus puts a large burden on the public health sector presenting an emerging health problem in the world ⁴². One of the therapeutic strategies to control hyperglycemia in diabetics is to avoid the hydrolysis of carbohydrates by the inhibition of α-amylase enzyme in order to slow down the intestinal absorption of glucose ⁴³. Alpha-amylase is the type of enzyme involved in the metabolism of carbohydrates i.e. hydrolysis of starch and disaccharides to glucose. In humans α-amylase expressed as two isoforms, secreted from Salivary glands and Pancreas respectively. These enzymes play an important role in digestion of polysaccharides like starch which is the main source of glucose in the human diet ^44-46. Human Salivary α-amylase (HSA) initiates the hydrolysis of α-(1, 4) glycosidic bonds in the dietary starch in to smaller oligosaccharides ⁴⁷. Further digestion of carbohydrates takes place in the gut under control of human pancreatic α-amylase (HPA) ⁴⁸.

Hence, hydrolysis of polymeric starch by HAS and HPA produces maltose, maltotriose and other oligomers, which are then degraded into glucose by α-glucosidase ⁴⁹. Inhibition of α-amylase results in delayed carbohydrate digestion and glucose absorption with attenuation of post prandial hyperglycemia ⁵⁰. The inhibitors of these enzymes are considered as the potential targets in the management of diabetes mellitus. Some synthetic antidiabetic drugs are available such as Acarbose, Miglitol and Voglibose but they are associated with gastrointestinal side effects such as adnominal pain, meteorism and diarrhea ^{51, 52}.

These side effects arise as a consequence of maltose fermentation which accumulates due to α-amylase inhibition. Therefore a non-carbohydrate based α-amylase inhibitor is expected to be better agent to restrain post prandial hyperglycemia since it would not lead to abdominal accumulation of maltose ⁵³. Many herbal products including several metals and minerals have been described for the case of diabetes mellitus. Plant products are frequently considered to be less toxic and significantly free from side effects than synthetic ones ⁵⁴. Inhibition of carbohydrate hydrolyzing enzymes by medicinal plants might be attributed to several possible factors such as encapsulation of starch by the fibers present in sample thus reduce the availability of starch to the enzymes or direct absorption of the enzyme on fibers that decrease enzyme activity ⁵⁵. Recently inhibition of α-amylase enzyme method became very good tool to estimate the antidiabetic potential of plant extracts ⁵⁶.

In the present study the concentrated and dried extracts Kandelia candel and Rhizophora apiculata were evaluated for antidiabetic activity by employing standard in vitro techniques (Alpha-amylase and glucose uptake assay using yeast model). In the Alpha-amylase inhibitory assay the known concentration (100µg) of different solvent extracts of Kandelia candel and Rhizophora apiculata were subjected to α-amylase inhibitory assay along with Acarbose as a standard. In case of Kandelia candel among five solvent extracts methanol extract showed very good antidiabetic activity on comparison with other tested extracts with Alpha-amylase inhibitory percentage 50.1907±0.33250 with IC₅₀ value 99.62µg/ml other remaining extracts i.e. chloroform extract, ethyl acetate extract, ethanol extract and aqueous extract showed percentage of inhibition 8.8481±0.46397, 13.3486±0.40362, 43.5545±0.80724 and 28.0701± 1.05970 respectively (Table 1).

TABLE 1: α -AMYLASE INHIBITORY ACTIVITIES AND IC₅₀ VALUES BY KANDELIA CANDEL EXTRACTS

Results are expressed as Mean±SE (n=3); **significant at the P < 0.01

**Correlation is significant at the 0.01 level (2-tailed) **, *Correlation is significant at the 0.05 level (2-tailed)*

FIG. 1: α -AMYLASE INHIBITORY ACTIVITIES OF KANDELIA CANDEL EXTRACTS

CE: Chloroform extract; EAE: Ethyl acetate extract; ME: Methanol extract; EE: Ethanol extract; AE: Aqueous extract; STD: Standard (Acarbose)

In case of Rhizophora apiculata the aqueous extract showed higher activity among all other extracts as well as methanol extract of Kandelia candel with percentage of inhibition 52.7078±0.66497 and IC₅₀ value observed to be 94.86µg /ml (Table 2). On comparison with standard acarbose methanol extract of Kandelia candel and aqueous extract of Rhizophora apiculata shown less activity i.e standard showed highest percentage of Inhibition 71.0907±0.67796 with IC₅₀ value 70.33 µg/ml (Table 2). In Glucose uptake in Yeast cells model the different solvent extracts of Kandelia candel and Rhizophora apiculata leaves at different concentrations (50µg-250 µg) are subjected to in vitro glucose uptake assay using yeast as model. The percentage of glucose uptake in yeast cells by the extract was compared with Metronidazole standard drug. In Glucose uptake assay both Kandelia candel and Rhizophora apiculata extracts and standard showed dose dependant manner of activity i.e. as the concentration of sample increased even the percentage of inhibition also increases. In case of Kandelia candel methanol extract shown higher activity than the remaining extracts with the percentage of inhibition 60.3143±0.56826 at 250µg concentration and it was less as compared to activity of standard.

TABLE 2: α-AMYLASE INHIBITORY ACTIVITIES AND IC₅₀ VALUES BY RHIZOPHORA APICULATA EXTRACTS

Results are expressed as Mean±SE (n=3); **significant at the P < 0.01.

**Correlation is significant at the 0.01 level (2-tailed)**, *Correlation is significant at the 0.05 level (2-tailed)*

FIG. 2: α -AMYLASE INHIBITORY ACTIVITIES OF RHIZOPHORA APICULATA EXTRACTS

CE: Chloroform extract; EAE: Ethyl acetate extract; ME: Methanol extract; EE: Ethanol extract; AE: Aqueous extract; STD: Standard (Acarbose)

In case of Rhizophora apiculata aqueous extract exhibited significant activity over all remaining tested extracts with percentage of inhibition 61.3667±0.73106 at 250µg concentration. Both plant extracts exhibited lesser activity on comparison with Metronidazole standard drug; results are shown in Table 3 and Table 4. The results indicate that methanol extract of Kandelia candel and aqueous extract of Rhizophora apiculata shown appreciable antidiabetic activity in performed in-vitro assays where as other tested extracts showed the least antidiabetic activity.

Plants are provided with secondary metabolites such as alkaloids, flavonoids, tannins, phenols and saponins which are also known as bioactive compounds and these bioactive compounds possess different biological activities which include antibacterial, antioxidant, anti-inflammatory, anticancer and antidiabetic and especially flavonoids already reported to treat type 2 diabetes ^57-60. In the recent years medicinal plants and their products have been investigated for potential benefits in the management of type 2 diabetes ^{61, 62}.

TABLE 3: PERCENTAGE OF GLUCOSE UPTAKE IN YEAST CELLS TREATED WITH KANDELIA CANDEL EXTRACTS

Results are expressed as Mean±SE (n=3); * significant at the P < 0.01.

Correlation is significant at the 0.01 level (2-tailed)**, Correlation is significant at the 0.05 level (2-tailed)*

FIG. 3: PERCENTAGE OF GLUCOSE UPTAKE IN YEAST CELLS TREATED WITH KANDELIA CANDEL EXTRACTS

TABLE 4: PERCENTAGE OF GLUCOSE UPTAKE IN YEAST CELLS TREATED WITH RHIZOPHORA APICULATA EXTRACTS

Results are expressed as Mean±SE (n=3); * significant at the P < 0.01.

Correlation is significant at the 0.01 level (2-tailed)**, Correlation is significant at the 0.05 level (2-tailed)*

FIG. 4: PERCENTAGE OF GLUCOSE UPTAKE IN YEAST CELLS TREATED WITH RHIZOPHORA APICULATA EXTRACT

CONCLUSION: In the present study in both in-vitro methods Kandelia candel and Rhizophora apiculata plant extracts showed antidiabetic activities but among tested extracts methanol extract of Kandelia candel and aqueous extract of Rhizophora apiculata exhibited higher antidiabetic activity over all extracts with good percentage of inhibition. Based on the present study results it can be used for the development of new pharmaceutical drugs for treatment and curing of diabetes and also this study shows that these extracts offer a safe method or supplement treatment strategy to control diabetes through its alpha amylase inhibition. However further comprehensive chemical and pharmacological investigation should be carried out to isolate the active compounds and appropriate elucidation of its mechanism of action and it helps in the development of new pharmaceuticals to treat diabetes mellitus.

ACKNOWLEDGEMENT: The authors would like to thank Department of Biotechnology and Microbiology, Karnatak University, Dharwad, Karnataka, India for providing necessary facilities and laboratory for doing this research work.

CONFLICT OF INTEREST: We wish to confirm that there are no known conflicts of interest associated with this publication

REFERENCES:

1. American Diabetes Association ADA: Dyslipidemia management in adults with diabetes. Diabetes Care 2004; 27: 68–71.
2. World Health Organization, WHO: Diabetes Mellitus. Available at:http://who.int/ mediacentre/factsheets/ fs312/ en/index.html 2013.
3. Maiti R, Jana D, Das UK, Ghosh D: Antidiabetic effect of aqueous extract of seed of Tamarindus indica in streptozotocin induced diabetic rats. J Ethnopharmacol 2004; 92:85–91.
4. Wadkar KA, Magdum CS, Patil SS, Naikwade NS: Antidiabetic potential and Indian medicinal plants. J Herbal Med Toxicol 2008; 2:45–50.
5. Kaushik G, Satya S, Khandelwal RK, Naik SN: Commonly consumed Indian plant food materials in the management of diabetes mellitus. Diabetes Metab Syndrome. Clin Res Rev 2010; 4: 21–40.
6. Skyler JS: Diabetes mellitus: pathogenesis and treatment strategies. Chemistry 2004; 47:4113–7.
7. Mir MS, Darzi MM, Khan HM, Kamil SA, Sofi AH, Wani A: Pathomorphological effects of alloxan induced acute hypoglycaemia in rabbits. Alexandria J Med 2013; 49: 343–53.
8. Akhtar MS, Iqbal J: Evaluation of the hypoglycaemic effect of Achyranthes aspera in normal and alloxan-diabetic rabbits. J Ethnopharmacol 1991; 31:49–57.
9. Ruffa MJ, Ferraro G, Wagner ML, Calcagno ML, Campos RH, Cavallaro L: Cytotoxic effect of Argentine medicinal plant extracts on human hepatocellular carcinoma cell line. J Ethnopharmacol 2002; 79:335–9.
10. Liu IM, Tzeng TF, Liou SS, Lan TW: Improvement of insulin sensitivity in obese Zucker rats by myricetin extracted from Abelmoschus moschatus. Planta Med 2007; 73:1054–60.
11. Schimmer BP, Parker KL: Adrenocorticotropic hormone; adrenocortical steroids and their synthetic analogs; inhibitors of synthesis and actions of adrenocortical hormones. In: Hardman JG, Limbard LE, Gilman AG, Goodman-Gilman A, editors. Goodman & Gilman’s the pharmacological basis of therapeutics. Mc Graw- Hill; p. 1658–9.
12. Grover JK, Yadav S, Vats V: Medicinal plants of India with anti-diabetic potential. Journal of Ethnopharmacology 2002; 81, 81–100.
13. Oboh G, Ademiluyi AO, Akinyemi AJ, Henle TH, Saliu JA, Schwarzenbolz U: Inhibitory effect of polyphenol rich extracts of jute leaf (Corchorus olitorius) on key enzyme linked to type 2 diabetes (alpha amylase and alpha glucosidase) and hypertension (angiotensin I converting) in vitro. Journal of Functional Foods 2012; 4, 450–458.
14. Djeridane A, Hamdi A, Bensania W, Cheifa K, Lakhdari I, Yousfi M: The in vitro evaluation of antioxidative activity, alpha glucosidase and alpha amylase enzyme inhibitory of natural phenolic extracts. Diabetes-Metabolic Syndrome and Clinical Research Review. http://dx.doi.org/10.1016/ j.dsx. 2013.10.007.
15. Kumar D, Gupta N, Ghosh R, Gaonkara RH, Pala BC: Alpha glucosidase and alpha amylase inhibitory constituent of Carexbaccans: Bio-assay guided isolation And quantification by validated RP-HPLC–DAD. Journal of Functional Foods 2013; 5: 211–218.
16. Yao X, Ling Zhu L, Chen Y, Tian J, Wanga Y: In vivo and in vitro antioxidant activity and a-glucosidase, a-amylase inhibitory effects of flavonoids from Cichorium glandulosum Food Chemistry 2013; 39: 59–66.
17. Kirtikar KR and Basu BD: Indian Medicinal Plants, (L.M. Basu, Allahabad), 2(1), 2004, 2015.
18. Barkat MA and Mujeeb M: Comparative study of Catharanthus roseus extract and extract loaded chitosan nanoparticles in alloxan induced diabetic rats. International Journal of Biomedical Research 2013; 04(12): 670-678.
19. Swathi Pothireddy: International Journal of Research and Development in Pharmacy and Life Sciences 2014; 3(3):1004-1010.
20. Rachala Vinod Kumar: Phytochemical constituents and biological activities of Homaliu zeylanicum Int J Pharm Bio Sci 2014; 5(3):176-182.
21. Siva Sankara Prasad K: Assessment of the analgesic and anti-inflammatory effect of Homalium zeylanicum. jgtps 2014; 5(3): 1886-1890.
22. Kumar dileep: Clinical evolution of Sarpagandha churna in the management of Yoshaapasmara vis-a-vis hysterical neurosisl. irjp 2014; 5(3):174-177.
23. Natava Rajesh: In -vitro and In-vivo Studies on the Antidiabetic Activity of Stem Bark of Homalium zeylanicum in STZ Induced Diabetic Rats. Asian Journal of Biochemical and Pharmaceutical Research 2014; 4(3):76-90.
24. Rollet B: Bibliography on mangrove research. London: UNESCO Paris Pub Information Retrieval Ltd 1981; 1600– 1975, p. 479.
25. Saxena H: A survey of the plants of Orissa (India) for tannins, saponins, flavonoids and alkaloids. Lloydia 1975; 38: 346-51.
26. Tiwari P, Rahuja N, Kumar R, Lakshmi V, Srivastava MN, Agarwal SC, et al: Search for antihyperglycemic activity in few marine flora and fauna. Indian J Sci Technol 2008; 1(5): 1-5.
27. Thangam TS, Kathiresan K: Mosquito larvicidal activity of mangrove plant extracts and synergistic activity of Rhizophora apiculata with pyrethrum against Culex quinquefasciatus. Int J Pharma 1997; 35, 1-3.
28. Williams LA: Rhizophora mangle (Rhizophoraceae) triterpenoids with insecticidal activity. Naturwissenschaften 1999; 86; 450-2.
29. Nurhanan MY, Asiah, O, Mohd Ilham MA, Siti Syarifah MM, Norhayati I, Lili Sahira H: Anti-proliferative activities of 32 Malaysian plant species in breast cancer cell lines. J Trop for Sci 2008; 20: 77-81.
30. Rahim AA, Rocca E, Steinmetz J, Kassim MJ, Ibrahim MS, Osman H: Antioxidant activities of mangrove Rhizophora apiculata bark extracts. Food Chem 2008; 107: 200-7.
31. Vinod P and Guruvayoorappan C: Anti-inflammatory and Anti-tumor Activity of the Marine Mangrove Rhizophora apiculata. J. Immunotoxicol 2014; 9: 341-452.
32. Sukardjo S: Forest Ecol Manag 1987; 20: 233–252.
33. Afidah AR, Rocca E, Steinmetz J, Kassim MJ, Adnan R, Ibrahim MS: Corros Sci 2007; 49: 402–417.
34. Motsei ML, Lindsey KL, Van Staden, J, Jager AK: J Ethnopharmacol 2003; 86: 235 –241.
35. School of Thai Traditional Medicine. Principle of Thai traditional Pharmacy. Bangkok: Prachettuphon wimolmungkalaramrajworawihorn Temple Press, 1981.
36. Traditional medicine association (Mahathaad Temple). A text book of Thai traditional pharmacy. Bangkok: Pithukaksorn Press, 1980.
37. Loo AY, Jain K and Darah I: Antioxidant and radical scavenging activities of the pyroligneous acid from a mangrove plant, Rhizophora apiculata. Food Chem 2006; 104: 300-307.
38. Kirtikar KR, Basu BD: Indian Medical Plants. Lalit Mohan Basu, Allahabad, India 1935; p. 2793.
39. Malik CP and Singh MB. Plant Enzymology and Histoenzymology, Kalyani Publishers 1980; New Delhi p. 278.
40. Cirillo VP: Sugar transport in psychrophilic yeast. J Bacteriol. 1963; 84: 485–491.
41. Li-xia Y, Tong-hua L, Zong-tao H, Juan-e L, Li-li W: Research progress on the mechanism of single-Chinese medicinal herbs in treating diabetes mellitus. Chinese Journal of Integrative Medicine 2011; 17: 235–240.
42. Guo LP, Jiang TF, Lv ZH, Wang YH: Screening -glucosidase inhibitors from traditional Chinese drugs by capillary electrophoresis with electrophoretically mediated microanalysis. J. Pharm. Biomed Anal 53 (2010): 1250–1253.
43. Manaharan T, Ling LT, Appleton D, Ming CH, Masilamani T, Palanisamy UD: Antioxidant and antihyperglycemic potential of Peltophorum pterocarpum plant parts. Food Chem 2011; 129: 1355-61.
44. Qin X, Ren L, Yang X, Bai F, Wang L, Geng P,  Bai G,  Shen Y: J. Struct. Biol 2011; 174, 196.
45. Butterworth PJ, Warren FJ, Ellis PR: Starch-Starke 2011; 63: 395.
46. Groot PC, Bleeker MJ, Pronk JC,  Arwert F, Mager WH, Planta RJ, Eriksson AW,  Frants RR: Genomics 1989; 5: 29.
47. Nikitkova AE, Haase EM, Scannapieco FA: Appl. Environ. Microbiol 2013; 79: 416.
48. Lehmann U, Robin F: Trends Food Sci. Technol 2007; 18: 346.
49. Li C, Begum A, Numao S, Park KH, Withers SG, Brayer GD: Biochemistry 2005; 44: 3347.
50. David SH, Bell MB: Type 2 diabetes mellitus: what is the optimal treatment regimen? The American Journal of Medicine 2004; 116: 23–29.
51. Lordan S, Smyth TJ, Soler-Vila A, Stanton C, Ross P: The -amylase and-glucosidase inhibitory effects of Irish seaweed extracts. Food Chem 2013; 141: 2170–2176.
52. Apostolidis E, Lee CM: In vitro potential of Ascophyllum nodosum phenolic antioxidant-mediated -glucosidase and -amylase inhibition. J. Food Sci. 2010; 75: 97–102.
53. Uchida R, Nasu A, Tokutake S, Kasai K, Tobe K, Yamaji N: Chem. Pharm. Bull. (Tokyo) 1999; 47: 187.
54. Dhandapani S, Vijayakumar R, Senthilkumar R, Nalini N: Hypolipidemic effect of Cuminum cyminum on alloxan-induced diabetic rats. Pharmacol Res 2002; 46 (3): 251–255.
55. Ou S, Kwok K, Li Y, Fu L: In vitro study of possible role of dietary fiber in lowering postprandial serum glucose. Journal of Agricultural and Food Chemistry 2001; 49: 1026–1029.
56. Abirami A, Nagarani G, Siddhuraju P: In vitro antioxidant, anti-diabetic,cholinesterase and tyrosinase inhibitory potential of fresh juice from Cit-rus hystrix and maxima fruits. Food Sci. Hum. Wellness 2014; 3:16–25.
57. Abdulrahman SH, Bushra HM: Flavonoids as alternatives in treatment of type 2 diabetes mellitus. Acad J Med Plant 2013; 1: 31-36.
58. Chitra J, Archana S, Padma K, Keerti G: Antidiabetic potential of flavonoids and other crude extracts of stem bark of Mangifera indica Linn: A comparative study. J Sci Inno Res 2014; 3(1): 21-27.
59. Tanko Y, Mohammed A, Musa KY and Eze ED: Evaluation of Effect of Ethanolic Leaf Extract of Mucuna pruriens on Blood Glucose Levels in Alloxan-Induced Diabetic Wistar Rats. Asian Journal of Medical Sciences 2012; 4(1): 23-28
60. Thenmozhi K, Jamuna S, Karthika K, Manian S, Paulsamy S, Chitravadivu C: HPTLC finger printing profile and evaluation of in vitro antidiabetic potential of medicinally important plant, Cassia obtusa (Caesalpiniaceae). J App Pharm Sci 2015; 5 (03): 073-079.
61. Kaur GS, Mukundan V, Wani and Kumar MS: Nutraceuticals in the Management and Prevention of Metabolic Syndrome. Austin J Pharmacol Ther 2015; 3:1063.
62. Mirmiran P, Bahadoran Z, and Azizi F: Functional foods-based diet as a novel dietary approach for management of types 2 diabetes and its complications: A review. World Journal of Diabetes 2014; 5(3): 267-281.
    

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    How to cite this article:

    Shettar AK and Vedamurthy AB: Evaluation of antidiabetic potential of Kandelia candel and Rhizophora apiculata- an in vitro approach. Int J Pharm Sci Res 2017; 8(6): 2551-59.doi: 10.13040/IJPSR.0975-8232.8(6).2551-59.

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