Research Article

Alcoholic leaf extract of Plectranthus amboinicus regulates carbohydrate metabolism in alloxan-induced diabetic rats
B. C. Koti, Aparna Gore, A. H. M. Thippeswamy, A. H. M. Viswanatha Swamy, Rucha Kulkarni ABSTRACT Objective: The present investigation was undertaken to explore the possible mechanisms of Plectranthus amboinicus leaf extract in alloxan-induced diabetic rats. Materials and Methods: Control and alloxan-induced diabetic albino rats received different treatments; orally control (vehicle), 200 mg/kg and 400 mg/kg of ethanol extract of Plectranthus amboinicus (PAEE) and 600 g/kg of glibenclamide (standard) for 15 days. At the end of the experiment, the animals were sacrificed and enzyme activities of carbohydrate metabolism were measured in the liver. Results: Diabetic control rats showed a significant elevation (P < 0.001) in fasting blood glucose on successive days of the experiment as compared with their basal values, which was maintained over a period of 2 weeks. Daily oral treatment with PAEE showed a significant reduction (P < 0.001) in the blood glucose levels on successive days of the experiment as compared with their basal values. The most pronounced antihyperglycemic effect was obtained with the dose of 400 mg/kg. PAEE shows a dose-dependent reduction in gluconeogenic enzymes like glucose-6-phosphatase and fructose-1,6-disphosphatase. After 15 days of treatment with PAEE, glycolytic enzymes like phosphoglucoisomerase resulted in a significant increase with a concomitant significant decrease in the activities of aldolase. On the other hand, glucose-6-phosphate dehydrogenase was significantly improved in diabetic rats on administration of PAEE; the 400 mg/kg dose of PAEE elicited a more potent effect compared with the 200 mg/kg dose. Conclusion: The results obtained in this study provide evidence of the antidiabetic activity of PAEE, mediated through the regulation of carbohydrate metabolic enzyme activities. KEY WORDS: Aldolase, alloxan, glucose-6-phosphate dehydrogenase, Plectranthus amboinicus

Department of Pharmacology, K. L. E. Universitys College of Pharmacy, Hubli, Karnataka, India Received: 25-06-2010 Revised: 23-10-2010 Accepted: 23-02-2011 Correspondence to: Prof. A. H. M. Thippeswamy, E-mail: tips_pg@yahoo.co.in

Introduction Diabetes mellitus is chiefly characterized by chronic hyperglycemia, with disturbances of carbohydrate, lipid and protein metabolism resulting from defects in insulin secretion, insulin action or both. Insulin deficiency and/or insulin resistance is associated with the pathogenesis of diabetic dyslipidemia and micro- and macrovascular complications. The commonly encountered acute and late diabetic
Access this article online Website: www.ijp-online.com
DOI: 10.4103/0253-7613.81520 Quick Response Code:

complications are already responsible for major causes of morbidity, disability and premature deaths in Asian countries. The underlying causes attributed to hyperglycemia ultimately result in oxidative stress, alterations in enzyme activities, protein glycosylation and several structural changes.[1] Alloxan acts as a diabetogenic owing to its ability to destroy the -cells of the pancreas possibly due to the free radical generation mechanism. Enzyme activities of gluconeogenesis have been shown to increase during the course of diabetes, with a simultaneous increase in the glycogenolytic and lipolytic pathways. Diabetes also has been accompanied with a decrease in the enzyme activities of the glycolytic and pentose phosphate pathways.[2] Plants have always been an exemplary source of drugs, and many of the currently available drugs have been derived directly or indirectly from them. Even the discovery of the widely used hypoglycemic drug metformin came from the traditional

286 Indian Journal of Pharmacology | June 2011 | Vol 43 | Issue 3

Koti, et al.: Effect of Plectranthus amboinicus in alloxan induced diabetic rats

approach of using Galega officinalis. There is a need to explore new drugs to modify the course of diabetic complications. In traditional system of medicine, Plectranthus amboinicus is used in the treatment of renal calculi, malarial fever, hepatopathy, chronic asthma, hiccough, helminthiasis and epilepsy.[3,4] The leaf extract of Plectranthus amboinicus contains chemical constituents like thymol, carvacrol, -caryophylline, -pinene, -pinene, camphor, cymene, camphene, -terpinene, p-cymene, limonene and -phellandrene, which are well known essential oils.[5] The plant is credited with antifungal,[6] antileptospiral,[7] rheumatoid arthritis[8] and nephroprotective activity.[9] However, the underlying mechanism by which the glucose level reduces needs to be investigated. The present investigation was undertaken to explore the antidiabetic effect and possible mechanisms of action of the leaf extract of Plectranthus amboinicus in alloxan-induced diabetic rats. Materials and Methods Animals Male Wistar albino rats weighing 150200 g were used. Animals were kept at room temperature (26 2C) for 1 week to acclimatize to laboratory conditions before starting the experiment. They were given free access to food and water ad libitum. Plant material Leaves of Plectranthus amboinicus were collected from Hubli, Karnataka, and were authenticated at the Department of Botany of H.S. Kothambri Science Institute, Hubli, and a voucher specimen has been deposited at the herbarium for further reference. Extract Preparation Powdered leaves (80 g) were macerated three times with 750 ml of 95% ethanol. The solvent was filtered and the hydroalcoholic extracts were concentrated in the Buchi rotavapour, which yielded about 8 g of the semisoild extract. This ethanol extract was kept in a refrigerator throughout the experimental duration. Induction of diabetes The model was developed by injecting the rats intraperitoneally with alloxan monohydrate dissolved in sterile saline (0.9% NaCl) at a single dose of 150 mg/kg. To avoid an early fatal hypoglycemic state, a 5% glucose solution was fed for 1 day to all the animals. The animals showing fasting blood glucose levels around 400450 mg/dl were selected for the study, determined at the 3rd day after alloxan administration. Experimental design A total of 30 rats were divided into five groups of six animals each. The daily oral treatments using fresh suspension of each drug were continued for 15 days,[10] with distilled water serving as the vehicle. Suspensions were prepared using 0.3% w/v sodium carboxy methylcellulose in distilled water and used for the experimental purpose. The volume of the suspension or vehicle was 5 ml/kg. All the groups received daily treatment orally between 08.00 and 09.00 h. Group I: Normal control (vehicle treated with 5 ml/kg of distilled water) Group II: Diabetic control (vehicle treated with 5 ml/kg of distilled water)

Group III: Diabetic rats + ethanol extract of Plectranthus amboinicus (PAEE) 200 mg/kg Group IV: Diabetic rats + PAEE 400 mg/kg Group V: Diabetic rats + glibenclamide 600 g/kg[11] At the end of the experimental period, the animals were deprived of food overnight and sacrificed under mild anesthesia. Blood and liver tissues were collected immediately for biochemical analyses. The liver tissue was dissected, washed in ice cold saline and blotted on filter paper. The liver was homogenized (10% w/v) with 0.1 M Tris-HCl buffer, pH 7.4 using a homogenizer. The tissue homogenate was then centrifuged at a speed of 3000 rpm for 15 min at room temperature and the supernatant was kept at 2C for further biochemical analysis. Biochemical Assays Blood was withdrawn retro-orbitally from the inner canthus of the eye with the help of a capillary tube under mild ether inhalation anesthesia at between 08.00 and 09.00 h. Blood samples were collected in Eppendroffs tubes and allowed to clot for 10 min. Serum was separated by centrifuging the samples at 3000 rpm for 10 min and stored in a refrigerator until the analysis. Glucose estimation along with their body weight was recorded in all the groups prior to treatment and 1 h after the respective treatment on the first, fourth, seventh, 10th and 15th days of the experiment. Blood glucose was determined by Trinders glucose oxidase method. Determination of glucose-6-phosphatase Glucose-6-phosphatase was assayed by the method described by Cori and Cori.[12] The reaction mixture contained 3 ml of potassium citrate (0.1 M, pH 6.8), 5 ml glucose-6phosphate (0.01 M, pH 6.8) and 2 ml homogenate. Blank was prepared using 5 ml of distilled water instead of glucose-6phosphate. Both the tubes were incubated at 30oC. After 1 h, 1 ml of 10% trichloro acetic acid was mixed and filtered. Inorganic phosphate was determined in 1 ml of the filtrate. Determination of fructose-1,6-diphosphatase Fructose-1,6-diphosphatase was determined by the method of Gancedo and Gancedo.[13] The reaction mixture at a final volume of 2 ml contained 1.2 ml of Tris-HCl buffer (0.1 M, pH 7.0), 0.1 ml of fructose-1,6-diphosphatase (0.05 M), 0.25 ml of MgCl2 (0.1 M), 0.1 ml of KCl (0.1 M), 0.25 ml of EDTA (0.001 M) and 0.1 ml of homogenate. Incubate the reaction mixture for 5 min at 37oC. One milliliter of 10% TCA was added and centrifuged. Inorganic phosphate was determined in 1 ml of the filtrate. Determination of phosphoglucose isomerase enzyme The activity of the Phosphoglucose isomerase (PGI) enzyme was measured according to the method described by Horrocks et al.[14] One milliliter of freshly prepared buffered substrate (glucose-6-phosphate in 10 M borate buffer, pH 7.8) was pipetted into two large test tubes named test and blank. 0.1 ml of the homogenate was added in both the tubes with or without incubation exactly for 30 min at 37oC in a water bath. At the same time, in the test tubes named standard and standard blank, 1 ml of the standard solution (fructose in 0.25% benzoic acid) and 1 ml of distilled water were added. Nine milliliters of chromogenic agent (HCl, resorcinol-thiourea reagent and distilled water, 7:1:1) was added to four tubes and placed in a 75C water bath for 15 min. The absorbance was
Indian Journal of Pharmacology | June 2011 | Vol 43 | Issue 3 287

Koti, et al.: Effect of Plectranthus amboinicus in alloxan induced diabetic rats

measured at 410 nm. The calculation of the activity is given by (Test - blank) / (Standard-blank) x 100 nmoles of fructose formed/min Determination of aldolase enzyme Aldolase enzyme was assayed by the method described by Sibley et al.[15] The reaction mixture contained 1 ml of Tris buffer (0.1 M, pH 8.6), 0.25 ml fructose-1,6-diphosphatase solution (0.05 M, pH 8.6) and 0.25 ml hydrazine solution (0.56 M, pH 8.6) in a test tube at 38oC. To this, sufficient water was added to make the volume to 2.5 ml. 0.5 ml of the homogenate was added to this mixture and the reaction was stopped after 30 min by the addition of 2 ml of 10% trichloroacetic acid. The blank was prepared in a similar manner, except that the fructose1,6-diphosphatase was added after trichloroacetic acid. The tubes were centrifuged. One milliliter of NaOH (0.75 N) was added in 1 ml of the supernatant. After 10 min, dinitrophenyl hydrazine was added and the tubes were placed in a 38oC water bath for 10 min. Seven milliliters of NaOH was added to give a total volume of 10 ml. The absorbance was read at 540 nm after 10 min of addition of alkali. The percentage transmission was plotted against mm3 Fructose-1,6-diphosphate (FDP) split/h. The value obtained was represented in mols as 22.4 mm3 of FDP = 1 mol of FDP = 2 mols of triose phosphate (glyceraldehyde) formed. Determination of glucose-6-phosphate dehydrogenase The activity of glucose-6-phosphate dehydrogenase was determined by the method of Ellis et al.[16] The incubation mixture contained 1 ml of Tris-HCl buffer (0.05 M, pH 7.5), 0.1 ml MgCl2 (0.1 M), 0.1 ml of nicotinamide adenine dinucleotide phosphate (0.1 M), 0.5 ml of phenazine methosulfate (0.1 M), 0.4 ml of dye solution and 0.1 ml of homogenate. The mixture was allowed to stand for 10 min at 37oC. The reaction was initiated by the addition of 0.5 ml of glucose-6-phosphate. Absorbance was read at 640 nm using water as a blank for 5 min at 1 min interval. The enzyme activity was expressed in units by multiplying the change in absorbance/min by the factor 6/17.6, the molar extinction co-efficient of the reduced co-enzyme. Determination of inorganic phosphate Inorganic phosphate was measured according to the method of Horwitt.[17] One milliliter of the filtrate was mixed with 8.75 ml of distilled water and 2.5 ml of solution E (15% trichloroacetic acid). After 5 min, the mixture was centrifuged and filtered. 1.25
Table 1:

ml of the supernatant was added to 12.5 ml of solution A (10% trichloroacetic acid) and 1.25 ml of solution B (1% ammonium molybdate). After 7 min, 1.25 ml of solution D (stannous chloride in HCl) was mixed. Readings were taken at 660 nm after 15 min. The standards representing different concentrations of phosphorus (2 mg, 4 mg, 6 mg, 8 mg and 10 mg) in 100 ml were prepared using KH2PO4 solution and processed in the same manner as the serum sample. The standard curve was plotted using concentrations of phosphorus (mg/100 ml) versus absorbance. Enzyme activities were obtained in the respective units by using a conversion factor. Statistical Analysis The results were expressed as mean S.E.M. The results obtained from the present study were analyzed using one-way ANOVA followed by Dunnetts multiple comparison tests. Data were computed for statistical analysis using the Graph Pad Prism Software. Differences between the data were considered significant at P < 0.05. Results The antihyperglycemic effect of graded doses of PAEE on fasting blood glucose is shown in Table 1. Diabetic control rats showed a significant elevation (P < 0.001) in fasting blood glucose on successive days of the experiment as compared with their basal values, which was maintained over a period of 2 weeks. Daily oral treatment with PAEE showed a significant reduction (P < 0.001) in blood glucose on successive days of the experiment as compared with their basal values. The most pronounced antihyperglycemic effect was obtained with a dose of 400 mg/kg. The activities of glucose-6-phosphatase and fructose1,6-diphosphatase enzymes were significantly increased (P < 0.001) in diabetic control rats as compared with control animals. Oral administration with PAEE reversed the values to normal (P < 0.01), as shown in Table 2. Enzymes like PGI and glucose-6-phosphate dehydrogenase showed a significant decrease (P < 0.001), whereas the aldolase enzyme exhibited a significant increase (P < 0.001) in their levels in diabetic control rats compared with normal control rats. The activities of PGI and glucose-6-phosphate dehydrogenase resulted in a significant increase (P < 0.001), with a significant decrease (P < 0.001) in the activities of aldolase in the PAEE-treated rats, as shown in Table 3.

Effect of PAEE (200 mg/kg and 400 mg/kg) on blood glucose level (mg/dl) in the control and experimental rats
Treatment Basal values After treatment 1st day 4th day 7th day 10th day 15th day Control 85.14 0.596 82.45 0.325 83.47 0.268 83.67 0.263 84.40 0.491 85.45 1.214 Diabetic control 384.4 0.9036 435.8 0.768*** 493.3 1.161*** 503.1 0.173*** 519.4 1.116*** 544.0 1.545*** PAEE 200 mg/kg 363.6 1.117 306.3 1.120*** 274.8 1.079*** 236.3 1.060*** 196.4 1.469*** 126.3 0.768*** PAEE 400 mg/kg 438.6 1.002 355.0 1.025*** 287.8 0.776*** 206.7 1.118*** 148.0 0.675*** 101.9 1.730*** GLB 426.6 1.215 365.9 1.135*** 283.3 0.703*** 206.5 0.809*** 135.0 0.744*** 95.18 1.413***

Values are given as mean SEM (n = 6); Values were statistically signicant at *P<0.05; **P<0.01; ***P<0.001; All the groups were compared with their basal values of the respective group; ns, non-signicance

288 Indian Journal of Pharmacology | June 2011 | Vol 43 | Issue 3

Koti, et al.: Effect of Plectranthus amboinicus in alloxan induced diabetic rats

Discussion Alloxan, a -cytotoxin, has demonstrated severe physiological and biochemical derangements of the diabetic state. The alloxan rats exhibited severe glucose intolerance and metabolic stress as well as hyperglycemia due to a progressive oxidative insult interrelated with a decrease in endogenous insulin secretion and release.[18] Several studies show that treatment with antioxidants might be an effective strategy for reducing diabetic complications due to disproportionate generation of free radicals. Studies focusing on the mechanisms of oxidative stress in diabetes have been carried out in order to develop a causal antioxidant therapy. In this study, diabetic rats exhibited a severe hyperglycemia. Treatment with PAEE has successively reduced the serum glucose level measured at the first, fourth, seventh and 10th days and at the end of the study both in normal and in diabetic rats. A decrease in the serum glucose level in normal rats was found to be an indication of hypoglycemic action of the PAEE treatment. The 400 mg/kg dose shows a consistent decrease in serum glucose. High levels of oxidative cytotoxicity have been linked to glucose oxidation, lipid abnormalities and non-enzymatic glycation of proteins, which contribute to the development of diabetic complications.[19] Increased glucooxidation results due to increased aldose reductase pathway activity leading to accumulation of sorbitol and fructose, NADP redox imbalances as well as alterations in signal transduction.[20] Carbohydrates, particularly glucose, are an important source of fuel for living organisms. The three major metabolic abnormalities that
Table 2: Effect of PAEE (200 mg/kg and 400 mg/kg) on glucose-6phosphatase (Ua/mg protein) and fructose-1,6-diphosphatase (Ub/ mg protein) in the control and experimental rats
Treatment Control Diabetic control PAEE 200 mg/kg PAEE 400 mg/kg GLB Glucose-6phosphatase 0.1652 0.02415 0.2678 0.01385## 0.2018 0.02327* 0.1810 0.009633** 0.1727 0.01039** Fructose-1,6diphosphatase 0.3597 0.01931 0.5837 0.1637# 0.4832 0.02573** 0.3950 0.01352*** 0.3760 0.01229***

a = moles of Pi liberated/min; b = moles of Pi liberated/h; Values are given as mean S.E.M. (n = 6) #P<0.001; ##P<0.01 compared with the control group; *P<0.05; **P<0.01; ***P<0.001 compared with the diabetic control group

contribute to hyperglycemia in diabetes mellitus are defective glucose-induced insulin secretion, increased hepatic glucose output and inability of insulin to stimulate glucose uptake in the peripheral target tissues. Pancreatic -cells appropriately alter their rates of insulin secretion in response to fluctuations in the levels of these calorigenic molecules, with glucose playing the dominant role in the regulation of insulin secretion. Ultimately, glycolysis and gluconeogenesis are two reciprocally regulated pathways to prevent wasteful glucose operation at the same time. Gluconeogenesis converts the carbohydrate as well as non-carbohydrate precursors to glucose or glycogen and clear metabolic products of the other organs. Glucose-6-phosphatase and fructose-1,6-disphosphatase are the important regulatory enzymes in the gluconeogenic pathway, which get altered during the course of uncontrolled diabetes. Glucose-6-phosphatase catalyzes the conversion of glucose-6-phosphate to glucose and, simultaneously, provides hydrogen, which binds with NADP+ in the form of NADPH and enhances the synthesis of fats from carbohydrates, i.e. lipogenesis.[21] The generation of fructose6-phosphate from fructose-1,6-diphosphate is catalyzed by fructose-1,6-diphosphatase, which determines whether or not a tissue is capable of resynthesizing glycogen from pyruvate triosephosphates.[22] The activities of these two enzymes are found to be increased in the diabetic control rats compared with the normal rats. Treatment with PAEE significantly decreases the activity of these two enzymes in the liver of diabetic rats. Conversely, glycolytic enzymes such as PGI and aldolase are prime enzymes in glucose utilization. The conversion of G6P to F6P is reversibly catalyzed by the enzyme PGI. Aldolase catalyzes reversible aldol condensation in glycolysis. As compared with the normal control values, the mean levels of hepatic PGI were seen significantly decreased, whereas the activity of hepatic aldolase was significantly increased in the diabetic controls.[23] Administration of PAEE to alloxan-induced rats resulted in an increased activity of serum PGI. A significant decrease in the activity of aldolase was found in diabetic rats treated with PAEE. Glucose-6-phosphate dehydrogenase catalyzes the conversion of glucose-6-phosphate to 6-phosphogluconate, with a simultaneous oxidation of NADP+ to NADPH. This maintains adequate levels of glutathione in its reduced form and helps to overcome oxidative stress.[24] The significant increase in the activity of glucose-6-phosphate dehydrogenase

Table 3: Effect of PAEE (200 mg/kg and 400 mg/kg) on phosphogluco isomerase (Uc/mg protein), aldolase (Ud/mg protein) and glucose-6-phosphate dehydrogenase (Ue/mg protein) in the control and experimental rats
Treatment Control Diabetic control PAEE 200 mg/kg PAEE 400 mg/kg GLB Phosphogluco isomerase 47.50 1.310 22.67 1.810# 34.17 2.128* 40.17 3.007*** 44.33 2.623*** Aldolase 0.14 0.0048 0.2477 0.0117# 0.2057 0.0094* 0.1762 0.0063*** 0.1612 0.0086*** Glucose-6-phosphate dehydrogenase 3.167 0.3997 1.100 0.1528# 2.367 0.2092* 2.750 0.2405** 3.000 0.4590***

c = nmoles of fructose formed/min; d = moles of glyceraldehyde formed/min; Values are given as mean S.E.M. (n = 6) #P<0.001 compared with the control group; *P<0.05, **P<0.01, ***P<0.001 compared with the diabetic control group

Indian Journal of Pharmacology | June 2011 | Vol 43 | Issue 3 289

Koti, et al.: Effect of Plectranthus amboinicus in alloxan induced diabetic rats

in PAEE 200 mg/kg- and 400 mg/kg-treated hyperglycemic rats suggests that the hydrogen shuttle systems and the redox state of the cell becomes more oxidized, which results in the increased formation of NADPH for increased utilization in lipogenesis and in turn activates the enzyme, as NADPH is a strong inhibitor of glucose-6-phosphate dehydrogenase.[25] A sequential metabolic correlation between decrease in the activity of enzymes of gluconeogenesis, increase in the activity of glycolytic enzymes, increased hydrogen shuttle reactions and normoglycemia stimulated by PAEE suggests the possible biochemical mechanism through which glucose homeostasis is regulated. The present study further reveals that this may be through an increase in insulin production very similar to the metformin-like action. In this study, in aloxanized rats, PAEE produced a significant and dose-dependent antidiabetic activity effect as compared with Trigonella foenum gracum, Eugenia jambolana and Ocinum sanctum beginning from the first day and progressing till the end of the 15th day.[26] In conclusion, treatment with PAEE (200 mg/kg and 400 mg/ kg) altered the metabolism in the liver of diabetic animals. Hepatic levels of G6Pase, FDPase, PGI and aldolase enzymes arrived to acceptable values after completion of treatment. Decreased gluconeogenesis and improved utilization of glucose by peripheral tissues suggests a metformin-like action of plant treatment.[27] Plectranthus amboinicus seems to have a promising value for the development of potent phytomedicines for diabetes, which may be due to the presence of flavonoids. Acknowledgment
The authors thank the Principal, K.L.E. Universitys College of Pharmacy, Hubli, India, for providing the necessary facilities to carry out the work.

9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. 21. 22. 23.

24. 25.

References
1. 2. Akpan HB, Adefule AK, Fakoya FA, Caxton-Martins EA. Evaluation of LDH and G6-PDH activities in auditory relay centers of streptozotocin-induced diabetic wistar rats. J Analytical Sciences 2007;1:21-5. Punitha SR, Rajendran K, Shirwaikar A, Shirwaikar A. Alcoholic stem extract of Coscinium fenestratum regulates carbohydrate metabolism and improves antioxidant status in streptozotocin-nicotinamide induced diabetic rats. Evid Based Complement Alternat Med 2005;2:375-81. Kirtikar KR, Basu BD. Indian medicinal plants. 2nd ed. Dehradun: International Book Distributors; 1975. p. 1971. Nadkarni AK. Indian Materia Medica. 2nd ed. Mumbai: Popular Prakashan; 1996. p. 371. Abdel-Mogib M, Albar HA, Batterjee SM. Chemistry of genus Plectranthus. Molecules 2002;7:271-301. Tempone AG, Sartorelli P, Teixeira D, Prado FO, Calixto IA, Lorenzi H, et al. Brazilian ora extracts as source of novel antileishmanial and antifungal compounds. Mem Inst Oswaldo Cruz 2008;103:443-9. Nirmala Devi K. In vitro antileptospiral activity of Plectranthus amboinicus (Lour) Spreng. Pharmaologyonline 2008;2:95-8. Chang JM, Cheng C, Hung L, Chung Y, Wu R. Potential use of Plectranthus

26.

3. 4. 5. 6. 7. 8.

27.

amboinicus in the treatment of rheumatoid arthritis. Evid Based Complement Altern Med 2007;23:1-6. Palani S, Raja S1, Naresh R, Senthil Kumar B. Evaluation of nephroprotective, diuretic, and antioxidant activities of Plectranthus amboinicus on acetaminopheninduced nephrotoxic rats. Toxicol Mech Methods 2010;20:213-21. Babu V, Gangadevi T, Subramonium A. Antihyperglycemic activity of Cassia kleinii leaf extract in glucose fed normal rats and alloxan-induced diabetic rats. Indian J Pharmacol 2002;34:409-15. Pari L, Sathesh MA. Antidiabetic activity of Boerhaavia diffusa L.effect on hepatic key enzymes in experimental diabetes. J Ethnopharmacol 2004;91:109-13. Cori GT, Cori CF. Glucose-6-phosphatase of the liver in glycogen storage disease. J Biol Chem 1952;199:661-7. Gancedo JM, Gancedo C. Fructose-1,6-diphosphatase, phosphofructokinase and glucose-6-phosphate dehydrogenase from fermenting and non fermenting yeast. Arch Microbiol 1971;76:132-8. Horrocks JE, Ward J, King J. A routine method for the determination of phosphogluco isomerase activity in body uid. J Clin Pathol 1963;16:248-51. Sibley JA, Lehninger AL. Determination of aldolase in animal tissues. J Biol Chem 1948;859-72. Ellis HA, Kirkma HN. A colorimetric method for assay of erythrocyte glucose-6phosphate dehydrogenase. Proc Soc Exp Biol Med 1961;106:607-9. Horwitt BN. Determination of serum phosphate by means of stannous chloride. J Biol Chem 1952;199:537-41. Szkudelski T. The mechanism of alloxan and streptozotocin action in -cells of the rat pancreas. Physiol Res 2001;50:536-46. Maritim AC, Sanders RA, Watkins JB. Diabetes, oxidative stress, and antioxidants: A review. J Biochem Mol Toxicol 2003;17:24-38. Anderson JW. Alterations in metabolic fate of glucose in the liver of diabetic animals. Am J Clin Nutr 1974;27:746-55. Krebs HA, Woodford M. Fructose 1,6-diphosphatase in striated muscle. Biochem J 1965;94:436-45. Cox MM, Nelson DL. Lehninger Principles of Biochemistry. 5th ed. New York: W. H. Freeman; 2008. p. 527-64. Arathi G, Sachdanandam. Therapeutic effect of Semecarpus and Cardium Linn, nut milk extract oncarbohydrate metabolizing and mitochondrial TCA cycle and respiratory chain enzymes in mammary carcinoma rats. J Pharm Pharmol 2003;55:1283-90. Oni GA, Johnson RA, Oguntibeju OO. Detecting patients with glucose-6phosphate dehydrogenase deciency. JIACM 2005;6:42-4. Eddouks M, Khalidi A, Zeggwagh NA, Lemhadri A, Michel JB, Burcelin R. An understanding mechanistic approach of hypoglycemic plants. In: Eddouks M, editor. Advances in phytotherapy research. Research Signpost Publishing House, Trivandrum; 2009. p. 109-28. Grover JK, Vats V, Rathi SS. Anti-hyperglycemic effect of Eugenia jambolana and Tinospora cardifolia in experimental diabetes and their effects on key metabolic enzymes involved in carbohydrate metabolism. J Ethnopharmacol 2000;73:461-70. Vats V, Grover JK, Rathi SS. Evaluation of anti-hyperglycemic and hypoglycemic effect of Trigonella foenum-graecum Linn, Ocimum sanctum Linn and Pterocarpus marsupium Linn in normal and alloxanized diabetic rats. J Ethnopharmacol 2002;79:95-100.

Cite this article as: Koti BC, Gore A, Thippeswamy A, Viswanatha Swamy A, Kulkarni R. Alcoholic leaf extract of Plectranthus amboinicus regulates carbohydrate metabolism in alloxan-induced diabetic rats. Indian J Pharmacol 2011;43:286-90.

Source of Support: Nil. Conict of Interest: None declared.

290 Indian Journal of Pharmacology | June 2011 | Vol 43 | Issue 3

Copyright of Indian Journal of Pharmacology is the property of Medknow Publications & Media Pvt. Ltd. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use.