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Pharmacologia
Year: 2013  |  Volume: 4  |  Issue: 3  |  Page No.: 197 - 207

Preventive and Therapeutic Efficacies of Benincasa hispida and Sechium edule Fruit’s Juice on Sweet-beverages Induced Impaired Glucose Tolerance and Oxidative Stress

Ashok Kumar Tiwari, Iragavarapu Anusha, Maddineni Sumangali, Domati Anand Kumar, Kuncha Madhusudana and Sachin Bharat Agawane    

Abstract: Aim: Increased consumption of sugar-sweetened beverages have been identified as one of the major sources responsible for increasing global epidemic of hyperglycemia, hyperglycemia induced oxidative stress and consequently, development of number of diabetic complications. This research explored preventive and therapeutic potentials of fruit juice of Benincasa hispida (BH) and Sechium edule (SE) in fructose and sucrose-sweetened beverages induced Impaired Glucose Tolerance (IGT), oxidative stress and IGT induced other metabolic disturbances in rats. Materials and Methods: Rats were orally administered fructose and sucrose sweetened-beverage prepared in water and respective fruit’s juice for one month. A group of rats were maintained for three months on daily single dose of sucrose. Three months sucrose-fed rats were later treated with fruit’s juice for fifteen days. Degree of IGT development was determined following Oral Glucose Tolerance Test (OGTT). Shape of plasma glucose concentration curve following OGTT, glycemic load, plasma Total Antioxidant Potential (TAOP), oxidative stress status, platelet aggregation activity and, Advanced Glycation End-products (AGEs) level in blood of rats were examined. Results: Fructose was more potent in inducing IGT than sucrose. Sucrose was more potent in decreasing plasma TAOP and inducing oxidative stress than fructose. BH juice prevented development of IGT induced by fructose and also significantly (p<0.05) reduced two hour glycemic load following OGTT. Three months continuous single dose administration of sucrose in rats induced development of IGT, increased two hour glycemic load (p<0.05) following OGTT, decreased TAOP (p<0.05), increased oxidative stress (more than twice) when compared with normal rats. Treatment of theses rats with juice of BH and SE accelerated normalization of oral glucose tolerance ability and decreased oxidative stress, platelet aggregation and accumulation of AGEs in sucrose induced IGT and oxidatively stressed rat. Conclusion: Our observations reveal that consumption of BH and SE juice may help reduce development of hyperglycemia and hyperglycemia induced development of complications by virtue of their antihyperglycemic and antioxidative stress potentials.

1,2,3,4. T2DM is emerging at epidemic rate in many parts of the world including developed nations like United States of America5 and developing countries like China6 and India7. Increasing evidences suggest that rise in consumption of sugar-sweetened beverages and food items enriched with high ratio of fructose is one of the important contributing factors exaggerating development of T2DM1. Although, the disaccharide sucrose is predominant sugar in most of the modern foods, the monosaccharide sweetener fructose is very common in a wide range of food products in some countries like USA8. Increased amount of fructose in diet is identified as an important causative factor responsible for increasing insulin resistance, decreasing insulin sensitivity and consequently, development of Impaired Glucose Tolerance2 (IGT). High-fructose diet is reported to induce oxidative stress by decreasing antioxidant defense9 and increasing free radicals generation10 Furthermore, postprandial hyperglycemic excursions (PPHGE) and hyperglycemic spikes even in non-diabetic individuals also increase oxidative stress11,12. Postprandial hyperglycemic spikes instigate redox imbalance in short term and lead to the development of complex chronic diseases in long-term exposure13.

Fruits and vegetables are being identified as potential readily available and cost-effective resources for development of new generation therapeutics for lifestyle related disorders14,15. Therefore, identification and study of fruits and vegetables that possess potentials to mitigate diet-induced IGT development, diminish oxidative stress level and correct IGT and oxidative stress induced biochemical and physiological imbalances may hold promise in reducing risks responsible for development of T2DM and diabetic complications.

Recommendations for inclusion of coarse cereals, fruits and vegetables in human diet have increased recently because such food items contain variety of potential biological antioxidants16,17 However, health benefits of polyphenols-rich dietary supplements18, dietary antioxidant supplements19 and antioxidant compounds from natural resources20 have been questioned recently for their prooxidant behaviors. Observations that consumption of polyphenolic-rich fruits increases antioxidant capacity of blood and counterbalances prooxidant effects induced by high carbohydrate and fat meals21 has been challenged by reports indicating that augmentation of blood antioxidant potential may be due to fructose-induced increase in serum uric acid level rather than polyphenols present in fruits-juice22,23,24. Although, uric acid possesses antioxidant properties, it is also a risk factor for several diseases25. In addition, some antioxidant-rich fruits augment hyperlipidemia26 and antioxidant-rich fraction of some cereals27 and dietary formulations28 display proglycemic activity. Vegetables are reported to be rich source of biological antioxidants17 and serve cost effective culinary item. However, high polyphenolic content in vegetable’s juice was observed recently to augment starch induced PPHGE15.

These reports therefore, warrant indiscriminate use of polyphenols-rich dietary antioxidant supplements and demand search of dietary materials that mitigates development of diet-induced glycemic burden and also reduces level of oxidative stress. Single dose oral administration of antioxidants rich juice of Benincasa hispida (BH) and Sechium edule (SE) was identified to mitigate starch induced PPHGE in rats15. Although, BH is extensively used in traditional oriental medicine for treatment of gastrointestinal problems, respiratory, heart and urinary diseases29 and SE advocated for treatment of arteriosclerosis, hypertension and inflammation30,31, effect of the juice of these fruits is not explored in sugar-sweetened beverages induced hyperglycemic conditions. In the present study, we report evaluation of preventive and therapeutic efficacy of BH and SE fruits juice on fructose and sucrose induced development of IGT and oxidative stress in rats.

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MATERIALS AND METHODS

Chemicals: 2, 2’-Azinobis-3-ethyl benzthiazoline-6-sulphonic acid diammonium salt (ABTS), Gallic acid, Rutin, Bradford and Folin-Ciocalteu reagents were purchased from Sigma-Aldrich chemicals (St Louis, MO, USA) and Platelets aggregation kits from Chronolog Corporation, USA. Kits for analysis of plasma glucose were obtained from Siemens Healthcare diagnostics Ltd, Frimley, Camberley, UK. Other chemicals like fructose, sucrose and potassium persulfate etc were purchased from Indian manufacturers.

Preparation of juice: Fruits of Benincasa hispida Thunb. Cogn. (Family Cucurbitaceae) and Sechium edule L. (Family Cucurbitaceae), were procured daily from local vegetable markets (Hyderabad, India). Fruits were washed thoroughly with water and cut into pieces. Juice was obtained by grinding in food-grade grinder and squeezed to maximum amount with clean muslin cloth.

Analysis of chemical components in juice: Chemical components like total polyphenols, flavonoids, anthocyanins and protein concentrations in fresh juice of these vegetables were analyzed as follows:

Total polyphenolic content: Total polyphenolic content in juice was measured using Folin-Ciocalteu reagent32. Briefly, fresh (25 μL) juice was diluted with 2.5 mL distilled de-ionized water followed by addition of 250 μL of Folin-Ciocalteu reagent (1 M) and 250 μL of Na2Co3 (20% w/v). Mixture was incubated at room temperature (60 min). Absorbance (765 nm) was recorded spectrophotometrically on microplate reader (SpectraMax plus384, Molecular Devices, Sunnyvale, CA). Total polyphenolic content was expressed as milligrams of Gallic Acid Equivalent (GAE)/mL juice
Total anthocyanins: Anthocyanins in juice were determined as described by Giusti et al.33. To 25 mM potassium chloride solution (pH 1.0) and 0.4 M sodium acetate buffer (pH 4.5), equal volumes of fresh juice were added. Absorbance was measured at 510 and 700 nm. Data was expressed using molecular extinction coefficient, molecular weight of anthocyanins and an absorbance of A = [(A510-A700 pH 1.0-(A510-A700) pH 4.5] as milligrams of anthocyanins per 100 mL juice
Total flavonoids: Total flavonoids content in samples was measured by mixing equal volume of vegetable’s juice with 2% AlCl3.6H2O in a 96 well micro plate34. Absorbance was recorded spectrophotometrically at 430 nm. Results were expressed as milligrams of rutin Equivalent (RE)/mL juice
Total protein content: Protein content in juices was determined using Bradford’s reagent35. Briefly, 10 μLjuice was mixed with 240 μL of Bradford reagent and absorbance was read at 595 nm spectrophometrically. Results were expressed as milligrams BSA Equivalent/mL juice

Animal experiments: Animal experiments were performed on adult male Wistar rats (170-210 g b.wt.). Institutional Animal Ethical Committee (CPCSEA Reg. No.97/1999, Government of India) approval for experimental protocol was obtained. Experiments with live animals were performed in compliance with relevant laws and institutional guidelines. Rats were housed three per cage in polyvinyl cages (with sterilized rice-husk bedding changed daily) in temperature-controlled room with 12 h light: dark cycle in institute’s animal house. Normal rat chow (Nutrimix Sp-1620, Nutrivet Life Sciences Pune, India) and water was provided ad libitum throughout experimental period.

On day one of experiment, rats were kept for overnight fasting. Next day forenoon blood was collected from retro-orbital plexus in EDTA-containing tubes. Basal plasma biochemical analysis (‘0’ h) was carried out with auto-blood analyzer instrument (Seimens Dimension Xplus analyser, Newark, USA).

Experimental design: Twenty percent fructose and fructose-equivalent sucrose solutions were prepared in distilled water and respective fruit’s juice routinely for oral administration to rats. Rats were grouped as follows:

Fructose group (Fructose feeding for one month, n = 6)
Sucrose group (Sucrose feeding for one month, n = 6)
BHF group (Fructose sweetened BH juice for one month, n = 6)
SEF group (Fructose sweetened SE juice for one month, n=6)
BHS group (Sucrose sweetened BH juice for one month, n=6)
SES group (Sucrose sweetened SE juice for one month, n=6)

Another set of eighteen rats were administered 40% sucrose solution (single dose) daily for three months. Sucrose feeding was withdrawn after three months feeding and rats were sub-divided in following groups:

Control (normal saline for fifteen days)
BH (treatment for fifteen days)
SE ((treatment for fifteen days)

A group of normal control animals (treated sham with normal saline) were also maintained throughout experimental period for comparative data analysis.

The dose of juice was decided as reported earlier15 and administered orally (7.5 mL kg-1 b.wt.) to animals once daily through gastric intubation (between 10-11 AM). Normal rat chow and water was provided ad libitum throughout experimental period. Daily administered dose of fructose and sucrose with and without juice was 1.5 and 3.0 g kg-1 b.wt., respectively.

Glucose tolerance test: Fasting blood was collected from retro orbital plexus in EDTA-containing tubes from overnight fasted rats after completion of experiments and plasma glucose levels were analyzed. Oral Glucose tolerance test (OGTT) was conducted by feeding glucose at dose of 2 g kg-1 b.wt. Blood was collected at intervals of 30, 60, 90 and 120th min post-glucose feeding and plasma glucose was analyzed on auto-blood analyzer instrument (Seimens Dimension Xplus analyser, Newark, U.S.A.). Two-hour postprandial glycemic load (area under the curve, AUC0-120 min) was calculated following trapezoidal rules15.

Glycemic load tolerance ability: Percentage change in glycemic load tolerance ability of rats was calculated as follows:

AUCsucrose 90 days represents AUC of rats fed sucrose for 90 days and AUCsucrose with drawn represents AUC of rats of either control or fifteen days juice treated groups after withdrawal of sucrose feeding.

Measurement of total antioxidant potential: Total antioxidant potential (TAOP) of plasma was estimated by measuring ABTS•+ cation scavenging activity. Plasma was deproteinised22 by mixing equal volume of acetonitrile and incubated for 2 min at 20°C and centrifuged for 10 minutes (9500xg, 4°C). ABTS•+ cation scavenging potential in deproteinised plasma was measured as reported earlier with slight modification15. Briefly, 100 mL stock solution of ABTS•+ (0.5 mM) was prepared by addition of 1 mL potassium persulfate [6.89 mM in PBS (pH 8.0)]. Mixture was stored in dark for 16 h. Deproteinised plasma (10 μL) was mixed with 190 μL of ABTS•+ in a 96-well microplate. Absorbance of decolorized ABTS•+ was measured at 734 nm after 10 min incubation in the dark on a microplate reader. Percentage ABTS•+ scavenging was calculated by applying following formula:

Determination of level of oxidative stress: Considering two-hour postprandial glycemic load (AUC0-120 min) as metabolic/oxidative stress inducer (prooxidant state) and plasma TAOP as available antioxidant defense, the ratio of AUC0-120 min and TAOP was calculated (AUC0-120 min/TAOP) as a measure representing level of oxidative stress in each group.

Analysis of whole blood platelet aggregation activity: Whole blood platelet aggregation activity was measured by impedance method36. Blood was collected in tubes containing 3.8% sodium citrate. One milliliter whole blood (diluted 1:1 with physiological saline) was transfer to plastic cuvette pre-heated at 37°C. Teflon-coated bar was added and electrode was placed in cuvette. Aggregation was induced by collagen (10 μg mL-1) and maximum amplitude was recorded on platelet aggregometer (Chronolog-Corporation USA, Model 700). Platelet aggregation was quantified as change in impedance and expressed in ohms.

Estimation of fluorescent AGEs: Plasma was deproteinised by mixing with equal volume of acetonitrile, incubated for 2 min (20°C) and centrifuged for 10 minutes (9500 x g, 4°C). Fluorescent AGEs in supernatant were assayed37 (excitation 370 nm, emission 440 nm) and expressed in fluorescent units.

Statistical analysis: One-way ANOVA followed by Bonferroni’s multiple comparison tests was applied to compare difference in study groups. Differences between pre-and post treatments were analyzed following Mann Whitney (two-tailed) test. Criterion for statistical significance was p<0.05. Statistical analysis was performed using GraphPad PRISM® Version 5.01 (GraphPad Software, Inc. California, USA) software.

RESULTS

Although the yield of juice from both vegetable fruits was similar, concentration of total polyphenols was more than twice and protein more than three times high in SE juice than in BH juice (Table 1). Total flavonoids content was recorded four times more in BH juice than in SE juice. Anthocyanins could not be detectable in these vegetables juice (Table 1).

Plasma glucose level remained high in one month fructose and sucrose fed rats over time. Development of IGT was higher in fructose than in sucrose administered rats. Furthermore, following OGTT, the glucose level in these animals did not return to the level of control group of animals over time (Fig. 1a). However, plasma glucose level returned to the normal level as in control group (120th min) when fructose and sucrose were administered dissolved in BH or SE juice (Fig. 1b, c). Although, the two-hour postprandial glycemic load (AUC0-120 min) following OGTT was significantly higher (p<0.05) in fructose or sucrose fed animals with or without juice when compared with control group (Fig. 1d), the BH juice significantly (p<0.05) mitigated glycemic burden in fructose and sucrose administered and SE in sucrose administered rats when compared with fructose fed rats (Fig. 1d).

One month continuous fructose and sucrose feeding significantly (p<0.05) decreased ABTS*+ cation scavenging activity of plasma (Fig. 2a). Drastic decrease in ABTS*+ cation scavenging activity was recorded in sucrose-fed animals. Both the vegetables juice significantly prevented decrease in ABTS*+ cation scavenging activity induced either by fructose or sucrose except in SEF group (Fig. 2a).

Level of oxidative stress in fructose and sucrose administered-rats was found increased (Fig. 2b). It was significantly (p<0.05) high in rats receiving sucrose with wide individual variations. Moderation in fructose induced oxidative stress was noticed in rats receiving BH and SE juice. Level of sucrose induced oxidative stress was significantly (p<0.05) mitigated (Fig. 2b) by BH (50%) and SE juice (70%).

Three months single-dose daily administration of sucrose solution significantly (p<0.05) increased fasting plasma glucose level of rats (89.7±6.9 vs. 98.2±6.4). Aggravation in IGT following OGTT was observed in rats’ withdrawn sucrose feeding for 15 days (Fig. 3a).


The postprandial glycemic spikes at 30th min in control group of rats were sharper than three months sucrose fed rats. Normalization in glucose tolerance was observed in rats treated with BH and SE juice for 15 days (Fig. 3a). The 2 h glycemic load in three months sucrose-fed rats was significantly higher (p<0.05) than normal rats (Fig. 3b). Three months sucrose feeding (Fig. 3b) induced comparable level of IGT induced by fructose feeding for one month (Fig. 1d). Glycemic load in control group rats was higher (9%) than sucrose fed rats. In comparison with control, BH and SE juice treatment for fifteen days significantly (p<0.05) improved two hour glycemic load (Fig. 3b).

One of the interesting finding in this study was that the glycemic load tolerance ability of rats decreased after withdrawal of sucrose feeding however, BH juice treatment significantly (p<0.05) improved it (Fig. 4). Although SE juice treatment also improved glycemic load tolerance ability of rats; statistical significance could not be observed (Fig. 4).

Decreased plasma TAOP by three months sucrose feeding was found significantly improved after withdrawal of sucrose feeding (Fig. 5a). Treatment of BH and SE juice augmented this improvement. However, statistical significance (p<0.05) could be found with SE juice treatment only. BH and SE juice treatment significantly decreased level of oxidative stress when compared with control (Fig. 5b).

Levels of fluorescent AGEs in plasma and collagen-induced platelet aggregation activity were recorded high in sucrose-fed control group of rats (Fig. 6).


In SE juice treated animals accumulation of AGEs was found significantly (p<0.05) less (Fig. 6a) however, platelet aggregation activity in whole blood (Fig. 6b) was recorded significantly less (p<0.05) in both the vegetables juice treated rats. SE juice was observed superior than BH juice in reducing both AGEs accumulation and platelet aggregation.

DISCUSSION

Plants make mixture of interacting compounds that provide important combination therapies by stimulating multiple therapeutic targets and impart clinical benefits beyond reach of single compound-based drugs38. Several mechanisms have been proposed for such therapeutic modalities. Phytochemical components interact with various targets to create a combination of effect.


One component might alter ability of another to reach its target, components bind separate sites on same target to create a combination effect and increase pharmacological action. Therefore, no single chemical component can be held responsible for activities displayed by complex plant mixtures because combination of whole might display more potential activity than sum of the parts39. These fundamentals have been the basis of oriental traditional medicines. In present study, the total polyphenols and protein concentration in SE juice was found more than BH juice whereas total flavonoids content in BH was high than in SE juice. Although, high polyphenolic content in vegetables juice is observed to augment starch induced postprandial glycemia, high protein content in vegetable’s juice is found to reduce starch-induced postprandial glycemic load in rats15. Therefore, variations observed in present study with regard to preventive and therapeutic potentials on various parameters might originate with synergistic and or antagonistic effect of different constituents present in juice of different vegetables.

Increased consumption of sugar-sweetened beverages in modern times results due to ample and affordable availability. Sweetened-beverages containing high ratio of fructose in particular, have been held responsible for increasing incidences of obesity40, alterations in carbohydrate and lipid metabolism41 and development of T2DM2. Our finding that pure-fructose (a monosaccharide) feeding for one month accelerated development of IGT and two hour glycemic load more than feeding of fructose-equivalent sucrose (disaccharide of glucose and fructose) following OGTT finds support with these observations.

The reason that fructose is more potent in aggravating development of IGT than sucrose may be that as a monosaccharide it is readily absorbed and metabolized by liver42. However, when administered in equivalent amount as sucrose, its availability for absorption is delayed because it requires action of disaccharide hydrolyzing enzyme α-glucosidase for its release. Furthermore, fructose and glucose follows different metabolic pathways. While glucose metabolism is regulated by main regulatory step of glycolysis controlled by phosphofructokinase, fructose by-passes this regulatory step and can continuously enter glycolytic pathway and adds to the uncontrolled production of glucose42. Furthermore, though fructose and sucrose solutions decrease ability of animals to tolerate glucose load, influence of fructose has been reported more than glucose43,44. Long terms sucrose feeding (three months) induced development of IGT up to the level induced by fructose in one month along with appearance of Impaired Fasting Glycemia (IFG). Interestingly, withdrawal of sucrose feeding after three months worsened glycemic load tolerance ability of rats without any treatment. These findings warrant sudden withdrawal of sugar intake without any precautionary measures.

Shape of plasma glucose concentration curve during OGTT has been recognized as an important measure to predict future risk of T2DM development. It has been demonstrated that subjects with normal glucose tolerance and IFG, whose plasma glucose concentration does not return to baseline after 60 min following OGTT, possess significantly higher risk of T2DM development45. Similar shape of plasma glucose curve following OGTT was observed in fructose and sucrose fed rats and demonstrates their IGT inducing potential. It was interesting to note that juice of both the vegetable fruits mitigated IGT inducing potential of fructose and sucrose. BH juice was more potent than SE in reducing fructose induced IGT development. Mechanism by which BH juice reduces fructose induced IGT development requires further investigation.

IGT and IFG represent intermediate state, which often progresses to overt diabetes within few year and is defined as pre-diabetes45. Pre-diabetes may be associated with increased risk of microvascular and macrovascular complications development45. Thus, reverting pre-diabetes state and preventing its development in to diabetes presents mammoth concern45. Three months sucrose feeding to rats induced both IFG and IGT. Fifteen days treatment of these rats with vegetables juice normalized shape of plasma glucose curve following OGTT and two hour glycemic load where as it remained higher in rats without any treatment. BH juice treatment was found significantly potent in improving glycemic load tolerance ability of rats.

Investigations report that fructose acts as pro-oxidant10,46 and induces oxidative stress by decreasing antioxidant defense and increasing free radicals generation10. However, glucose has been observed to act as pro-oxidant by enhancing pro-oxidant enzyme’s activity47. ABTS•+ cation has been successfully utilized to determine free-radical scavenging/antioxidant potential of plasma48. Utilizing ABTS•+ cation scavenging activity as a measure of plasma TAOP; we found that both fructose and sucrose significantly decreased plasma TAOP, sucrose being more potent than fructose. The intense decrease in antioxidant potential of plasma by sucrose may be due to cumulative pro-oxidant and oxidative stress inducing effect of fructose and glucose units present in disaccharide. Both the vegetable’s juice offered significant protection against sucrose induced decrease in plasma TAOP. SE offered better protection than BH. BH mitigated fructose induced decrease in plasma TAOP significantly. Withdrawal of sucrose feeding significantly improved plasma TAOP. Although, more increase in plasma TAOP by BH and SE treatment was noticed it was not significantly higher than untreated control rats. It still remains an open question that withdrawal of sucrose feeding decreased glycemic load tolerance ability of rats, TAOP of animals increased significantly.

Postprandial period is a pro-oxidant state. Postprandial period is also a time of active oxidative metabolism and generation of free radicals. Generation of pro-oxidant environment or decrease in antioxidant defense represents level of oxidative stress. An imbalance between oxidants generation and antioxidant defense in favor of oxidants potentially leading to physiological dysfunction and/or biomolecular damage is referred to as oxidative stress21. Utilizing these parameters, for example, two hour glycemic load as a state inducing pro-oxidant environment and TAOP as available antioxidant defense, we obtained oxidative stress level in experimental animals. Oxidative stress level in fructose-fed rats was moderately high than control group rats however; it was significantly high in sucrose-fed animals. These results are in accordance with the earlier observations that fructose-rich diet induces moderate oxidative stress49.BH and SE juice moderated development of oxidative stress induced by fructose however, offered significant protection against sucrose induced oxidative stress development. Similar results were obtained when rats were fed sucrose for three months. Both BH and SE juice treatment significantly accelerated decrease in oxidative stress level than rats treated as placebo.

Hyperglycemia is known to potentiate platelet activation50 and increase generation of advanced glycation end products (AGEs). These processes are further amplified under oxidative stress conditions12. Additionally, increased concentrations of AGEs further exacerbate platelet aggregation51. Altered platelet function is prevalent in diabetes and participates in pathogenesis of diabetic vascular complications by promoting microthrombus formation52.Both of these abnormalities were detected in three months sucrose fed rats. Therapies possessing antihyperglycemic and antioxidative properties therefore might reduce development of these complications. Whole blood platelet aggregation assay is a model more closely related to physiological conditions. Therefore, inhibition of whole blood aggregation would suggest antithrombotic potential of a therapy36. Juice of BH and SE significantly reduced collagen induced platelet aggregation in hyperglycemic blood. SE juice was found better in reducing collagen induced platelet aggregation and AGEs formation than BH juice. Presence of both antihyperglycemic and antioxidative activities in juice of BH and SE may be responsible for antiplatelet aggregation and anti-AGEs activities in this study.

CONCLUSION

Sugar-sweetened beverages are potent inducer of IGT, IFG, oxidative stress and risk factor responsible for development of hyperglycemia induced complications. Juice of BH and SE may offer protective and therapeutic measures against physiological and biochemical imbalances induced by increased intake of sugar-sweetened beverages by virtue of presence of multiple preventive and therapeutic activities. This report provides for the first time evidence that consumption of raw juice from vegetables may help ameliorate sweetened-beverages induced metabolic disturbances and impart health benefits in IGT individuals.

ACKNOWLEDGMENTS

Authors thank D’IICT for constant support and encouragement. This research was supported in part from Grant MLP-0001 (CSIR, New Delhi). Authors declare no conflict of interest, financial or otherwise.

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