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. 2011 May 5;50(3):528–534. doi: 10.1007/s13197-011-0367-3

A multigrain protein enriched diet mitigates fluoride toxicity

Rupal A Vasant 1, Narasimhacharya Amaravadi VRL 1,✉
PMCID: PMC3602553  PMID: 24425948

Abstract

Fluorosis is a major health problem in many parts of the world. The present work focuses on investigating the utility of nutrient and antioxidant rich grains- ragi, jowar, bajra, maize in formulation of basal, high carbohydrate low protein and low carbohydrate high protein diets in mitigating fluoride toxicity. Exposure to fluoride through drinking water not only significantly increased plasma glucose and lipid profiles, but also elevated both hepatic and renal lipid peroxidation, hepatic lipid profiles and G-6-Pase activity with a reduction in plasma HDL-C, hepatic glycogen content, hexokinase activity and antioxidant status. Even though basal and high carbohydrate diets did not significantly alter plasma glucose, lipid profiles in fluoride administered animals, protein enriched multigrain diet significantly decreased plasma glucose and lipid levels. However, the multigrain basal and high carbohydrate diets influenced the hepatic glycogen, lipid profiles, hexokinase and G-6-Pase activities, hepatic and renal lipid peroxidation and antioxidant status though not as significantly as that of multigrain diet enriched with protein. Thus the results of the present study indicate that both a multigrain diet rich in nutrients and antioxidants, and fortified with protein is useful in mitigating the fluoride toxicity.

Keywords: Antioxidants, Fluorosis, Lipid peroxidation, Oxidative stress, High protein diet

Introduction

Fluoride when present in higher concentrations in natural resources becomes a potential environmental and health hazard. People have long been exposed to fluoride through drinking water, food and air (Ando et al.1998). About 66.62 million people in India are at a risk for fluorosis due to high water- borne fluoride. In Indian state of Gujarat on the west coast, twenty four of its twenty six districts are reported to have high fluoride content in their water sources (Susheela 2007). Chronic exposure to fluoride is reported to cause hyperglycemia, hypercholesterolemia, hyperphospholipidemia and hypertriacylglycerolemia in experimental animals (Shashi 1992; Chlubek et al.2003; Grucka-Mamaczar et al.2004; Rupal et al.2010). Moreover, long term exposure to high fluoride content in early developmental stages was shown to increase the oxidative stress in blood and decreases the antioxidant defense systems in liver with declined activities of SOD, CAT, GPX, GSH, GST and TAA (Shivarajashankara et al.2003; Shanthakumari et al.2004). Besides the techniques being used to defluoridate water alternative methods are needed to ameliorate the toxic symptoms caused by fluoride. For instance, administration of vitamins C, D and E alone and in combination was reported to mitigate the fluoride toxicity in rats (Verma and Guna-Sherlin 2002). Dietary supplementations with calcium and aluminium salts, magnesium metasilicate (serpentine) and borates have also been shown to reduce the intensity of fluoride toxicity to some extent (Wagner and Muhler 1960; Jowsey and Riggs 1978; Reddy et al.1985). Synthetic diets incorporating casein, starch, salt and vitamin mixtures without the use of other ingredients such as cereal/millet flour, have been shown to ameliorate fluoride toxicity (Chinoy and Mehta 1999; Chinoy et al.2005a, b, 2006) and this effect was ascribed to the protein fraction of the diets.

Since protein, calcium, vitamins C, E, and antioxidants are basically obtained from plant/animal sources, research in the area of nutrition is now being focused on natural sources for formulating ‘healing diets’. Herbal or natural products are being increasingly investigated for their role in reducing the effects of fluoride toxicity for e.g., tamarind fruit pulp supplementation increased urinary excretion of fluoride while decreasing the retention of fluoride in bone (Khandare et al.2000, 2002, 2004; Ekambaram et al.2010). Limonia fruit powder supplementation reduced hepatic and renal tissue lipid peroxidation with a concomitant increase in the antioxidant status (Rupal and Narasimhacharya 2011). The seed and bark extracts of Moringa oleifera and Terminalia arjuna have also been shown to reduce fluoride induced toxicity (Stanely et al.2002; Ranjan et al.2009; Sinha et al.2007; Ghosh et al.2008). A 43 kD protein isolated from the leaves of the herb Cajanus indicus appears to play a protective role in fluoride induced oxidative stress (Manna et al.2007). Additionally, administration of black tea and black berry juice were found to be useful in reducing the effects of fluoride in laboratory animals (Verma et al.2007; Hassan and Yousef 2009). The present study deals with the formulation of diets using millets and cereals- Pennisetum typhoideum (Bajra), Eleusine coracana (Ragi), Sorghum vulgare (Jowar) and Zea mays (Maize) which are well-known for their use as foods in many parts of India. These are main sources of energy in Indian diets as the millets and cereals are important sources of several nutrients such as protein, calcium, iron, vitamin B-complex and fiber and these are the preferred millets and cereals in dry and drought prone areas. Millets including ragi are rich in minerals and fiber and cereals generally have low fat content. The carbohydrate content of these grains ranges from 66–72 g%, the protein content varies from 7.3 to 11.6 g% thus making these grains rich in carbohydrates and proteins besides being rich in amino acids (Gopalan et al.2004). These grains are also rich sources of polyphenols with a high antioxidant capacity (Sreeramulu et al.2009). Thus this study was conceived to address the problem of fluorosis by using an alternative ‘multi-grain’ diet to investigate if it can be a stand-alone diet or as a supplementary diet to treat fluorosis.

Materials and methods

Animals

Adult female Albino rats (Charles Foster) weighing 150–200 g were used for the present investigation. Animals were kept in polypropylene cages with ad libitum access to water (26 ± 2 °C, humidity 60%). The care and procedures adopted for the present investigation were in accordance with the approval of Institutional Animal Ethics Committee (MoEF/CPCSEA/Reg.337).

Experimental protocol

After a 10-day adaptation period, 30 animals were divided into 5 groups of 6 animals each as: NC-normal control with commercial diet; FC- fluoride control- 100 ppm NaF through drinking water and fed commercial diet, F Basal—fluoride treated animals were given basal diet, F HCLP—fluoride treated animals were fed with high carbohydrate low protein diet, F HPLC—fluoride treated animals were given high protein low carbohydrate diet. The commercial diet contained carbohydrates (55.67 g%), protein (22.12 g%), fat (4.06 g%), minerals (5.64 g%) and fiber (3.76 g%). The carbohydrate, protein, fat, mineral and fiber contents of the commercial diet were replicated approximately using the cereals and millets for basal diet preparation. The composition of the formulated diets is given in Table 1.

Table 1.

Composition of formulated diets

Cereals and Millets Basal (gm%) High carbohydrate low protein (gm%) High Protein low carbohydrate (gm%)
Bajra 20.0 20.0 16.25
Ragi 20.0 20.0 16.25
Jowar 20.0 20.0 16.25
Maize 20.0 20.0 16.25
Corn starch 4.0 16.0 –
Casein 12.0 – 31.0
Refined oil 4.0 4.0 4.0
Carbohydrate (gm%) 59.66 70.66 45.04
Protein (gm%) 20.08 8.08 39.26
Fat (gm%) 7.34 7.34 5.91
Minerals (gm%) 1.62 1.62 1.28
Fiber (gm%) 1.82 1.82 1.46
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At the end of the treatment period (4 weeks), animals were fasted overnight and sacrificed under mild ether anesthesia. Blood was collected by cardiac puncture and plasma was separated by centrifugation. Liver and kidney tissues were excised and, both plasma and tissues were kept frozen until analyzed.

Biochemical analyses

Plasma glucose and lipid profiles

Plasma glucose levels were measured by o-toluidine method with a glucose kit (Eve’s Inn Diagnostics, Baroda, India). Plasma total lipid (TL) content was estimated by sulphophosphovanillin method (Frings et al.1972). Plasma total cholesterol (TC), HDL cholesterol (HDL-C) and triglyceride (TG) contents were estimated with cholesterol and triglyceride kits respectively (Eve’s Inn Diagnostics, Baroda, India). Low-density lipoprotein cholesterol (LDL-C = TC−HDL-C−Tg/5), very low-density lipoprotein cholesterol (VLDL-C = Tg/5), and atherogenic index (AI = TC/HDL-C) were calculated (Friedewald et al.1972).

Hepatic glycogen and lipid profiles

Hepatic glycogen was extracted with 30% KOH and the yield was determined by anthrone- sulphuric acid method (Seifter et al.1950). The hepatic TL was extracted in chloroform: methanol (2:1) mixture (Folch et al.1957) and were estimated by gravimetric analysis. The same extract was used for the estimation of TC and TG contents by respective kits (Eve’s Inn Diagnostics, Baroda).

Hepatic hexokinase and G-6-Pase activities

The hepatic hexokinase (EC 2.7.1.1) was determined based on reduction of NAD+ through a coupled reaction with glucose-6-phosphate dehydrogenase (Brandstrup et al.1957). Glucose-6-phosphatase (EC 3.1.3.9) activity was determined by measurement of inorganic phosphate liberated from glucose-6-phosphate (Baginsky et al.1974).

Hepatic and renal lipid peroxidation and antioxidant profiles

The hepatic and renal lipid peroxidation (malondialdehyde concentration) was determined by the thiobarbituric acid (TBA) assay (Ohkawa et al.1979). Total ascorbic acid was estimated using 2, 4-dinitrophenyl hydrazine reagent (Schaffert and Kingsley 1955). Superoxide dismutase (SOD; EC 1.15.1.1) was measured using the nitroblue tetrazolium reduction method (Kakkar et al.1984). Catalase (CAT; EC 1.11.1.6) was assayed spectrophotometrically as decomposition of H2O2 at 240 nm according to the method described by Aebi (1974). Assay of glutathione peroxidase (GPx; EC 1.11.1.9) was based on GSH consumption as described by Flohe and Gunzler (1984). Reduced glutathione (GSH) was measured by reduction of DTNB as described by Jollow et al. (1974).

Statistical evaluation

Data are presented as mean±SEM. One-way analysis of variance (ANOVA) with Tukey’s significant difference post hoc test was used to compare differences among groups. Data were statistically handled by Graph Pad Prism 3.0 statistical software. P values < 0.05 and < 0.001 were considered statistically significant.

Results and discussion

High protein diets have been shown to be beneficial in maintenance of basal triglycerides, glucose, leptin and plasma insulin concentrations and do not produce any adverse effects on both renal and hepatic functions and on oxidative stress (Lacroix et al.2004). Lacroix et al. (2004) also proposed that the conversion of amino acids to glucose upon feeding high protein diet brings about a negative metabolic effect on the liver with regard to the balance in glycolysis/gluconeogenesis through alterations in the activities of key enzymes of glucose metabolism. The high protein low carbohydrate diets have also been shown to lower blood glucose levels post-prandially in type 2 diabetic individuals (Gannon et al.2003) and reduce serum trialcylglycerol, increase HDL- cholesterol and reduce blood pressure (Layman et al.2008). Besides, the protein supplemented diets have also been reported to accelerate fluoride metabolism and reduce the absorption and toxicity of fluoride by increasing its excretion (Boyde and Cerklewski 1987; Wang et al.1994; Chinoy and Mehta 1999; Chinoy et al.2006). Interestingly, dietary calcium decreases intestinal absorption of fluoride (Chinoy et al.1993); protein in the diet even while enhancing fluoride absorption does not favor its retention leading to its rapid excretion (Boyde and Cerklewski 1987). On the other hand, a low protein diet appeared to aggravate fluoride toxicity causing bone fragility (Reddy and Srikantia 1971) and a significant reduction in the activities of enzymatic and non- enzymatic antioxidants—SOD, CAT, GSH-Px, reduced ascorbic acid and GSH (Chinoy et al. 2005a; b). These observations clearly indicate that the dietary components- protein and calcium play a major role in reducing the fluoride load in the body and help mitigate fluoride toxicity.

Fluoride administered animals exhibited a significant increase in plasma glucose (98.22%) and lipid profiles TL, TC, TG, LDL-C, VLDL-C and AI (40.47, 37.25, 28.62, 155.73, 28.58 and 93.30% respectively) with a reduction in HDL-C contents (28.77%) when compared to the controls. Basal and high carbohydrate diets fed animals did not exhibit significant variations in plasma glucose and lipid profiles (Table 2). The protein enriched multigrain diet significantly decreased the plasma glucose and lipid profiles with a concomitant increase in HDL-C levels. Thus it appears that the protein fraction of the diet is important in maintenance of both plasma glucose and lipid profiles even in fluoride exposed animals indicating its usefulness in ameliorating fluoride toxicity.

Table 2.

Effect of diets on plasma glucose and lipid profiles

Parameters Control Fluoride control F Basal F HCLP F HPLC
Glucose (mg dl-1) 98.7 ± 0.23 195.7 ± 0.28a (+98.2) 194.1 ± 0.25a (−0.8) 197.4 ± 0.33a (+0.8) 146.7 ± 0.50ab (−25.06)
TL (mg dl-1) 323.4 ± 1.43 454.3 ± 2.10a (+40.4) 437.6 ± 1.36ab (−3.6) 449.7 ± 1.25a (−1.01) 391.2 ± 1.07ab (−13.8)
TC (mg dl-1) 110.7 ± 0.90 152.06 ± 0.86a (−37.2) 151.1 ± 0.53a (−0.5) 156.7 ± 0.69a (+3.08) 111.6 ± 0.89b (−26.6)
TG (mg dl-1) 69.8 ± 0.41 89.7 ± 1.15a (+28.6) 85.5 ± 0.31ab (−4.7) 93.2 ± 0.70a (+3.8) 69.4 ± 0.53b (−22.6)
LDL-C (mg dl-1) 35.3 ± 0.93 90.3 ± 0.98a (+155.7) 89.9 ± 0.48a (−0.4) 94.4 ± 0.61a (+4.5) 41.7 ± 0.95ab (−53.7)
VLDL-C (mg dl-1) 13.9 ± 0.08 17.9 ± 0.23a (+28.5) 17.06 ± 0.05ab (−4.9) 18.6 ± 0.14a (+3.9) 13.8 ± 0.11b (−22.6)
HDL-C (mg dl-1) 61.5 ± 0.31 43.8 ± 0.26a (−28.7) 44.1 ± 0.18a (+0.8) 43.6 ± 0.26a (−0.3) 55.9 ± 0.14ab (+27.6)
AI (mg dl-1) 1.7 ± 0.01 3.4 ± 0.02a (+93.3) 3.4 ± 0.01a (−1.1) 3.5 ± 0.02a (+3.4) 1.9 ± 0.02ab (−42.6)
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Figures in the parenthesis indicate% increase or decrease. Significant at P < 0.05, 0.001; a indicates the comparison with control group and b indicates the comparison with fluoride control group (n=6)

Exposure to fluoride through drinking water decreased the hepatic glycogen content (50.73%) and increased hepatic TL, TC and Tg levels. When HPLC diet was given, the hepatic glycogen content increased significantly (59.92%) and TL, TC and Tg contents decreased (32.58, 44.79 and 26.62% respectively). However, basal and HCLP diets did not cause significant variations in hepatic glycogen and lipid profiles (Table 3). Administration of fluoride through drinking water significantly increased the hepatic G-6-Pase activity (187.11%) and reduced hexokinase activity (40.55%) and, HPLC diet reversed this trend (G-6-Pase activity by 36.44%; increased hexokinase activity by 45.79%). However, no significant differences were found either in basal or HCLP fed animals when compared to fluoride intoxicated animals (Table 4). These observations clearly implicate the role of dietary proteins in influencing the hepatic carbohydrate and lipid metabolism in fluoride intoxicated rats in perhaps a similar manner as in diabetes (type 2) and adiposity (Gannon et al.2003; Lacroix et al.2004; Layman et al.2008).

Table 3.

Effect of diets on hepatic glycogen and hepatic lipid profiles

Groups Control Fluoride control F Basal F HCLP F HPLC
Glycogen (mg gm-1) 20.4 ± 0.02 10.08 ± 0.03a (−50.7) 10.1 ± 0.04a (+0.9) 10.2 ± 0.04a (+1.6) 16.1 ± 0.02ab (+59.9)
TL (mg gm-1) 29.1 ± 0.15 42.3 ± 0.10a (+45.01) 41.6 ± 0.14a (−1.6) 41.4 ± 0.11a (−1.9) 28.5 ± 0.10ab (−32.5)
TC (mg gm-1) 2.02 ± 0.01 3.8 ± 0.02a (+90.09) 3.7 ± 0.01a (−1.5) 3.8 ± 0.04a (−0.5) 2.1 ± 0.02b (−44.7)
TG (mg gm-1) 11.8 ± 0.34 19.1 ± 0.26a (+61.08) 19.8 ± 0.25a (+3.5) 19.6 ± 0.29a (+2.8) 14.03 ± 0.52ab (−26.6)
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Figures in the parenthesis indicate% increase or decrease. Significant at P < 0.05, 0.001; a indicates the comparison with control group and b indicates the comparison with fluoride control group (n=6)

Table 4.

Effect of diets on hepatic hexokinase and G-6-Pase activities

Groups Control Fluoride control F Basal F HCLP F HPLC
Hexokinase (U mg-1 protein min-1) 7.2 ± 0.03 4.2 ± 0.03a (−40.5) 4.4 ± 0.02a (+4.2) 4.7 ± 0.02ab (+11.4) 6.2 ± 0.10ab (+45.7)
Glucose-6-phosphatase (U mg-1 protein min -1) 0.194 ± 0.003 0.557 ± 0.007 a (+187.1) 0.554 ± 0.005a (−0.5) 0.550 ± 0.007a (−1.2) 0.354 ± 0.004ab (−36.4)
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Figures in the parenthesis indicate% increase or decrease. Significant at P < 0.05, 0.001; a indicates the comparison with control group and b indicates the comparison with fluoride control group (n=6)

Cellular oxidative stress is a result of imbalance between the production of reactive oxygen species and the protective antioxidant mechanisms (Halliwell and Gutteridge 1999). It has been shown that chronic exposure to fluoride increases lipid peroxidation with simultaneous reduction in the antioxidant enzymes (Shanthakumari et al.2004). Ascorbic acid is an important antioxidant that helps elimination of reactive oxygen species and reduces the oxidative stress (Oguntibeju 2008). Superoxide dismutase (SOD) is an enzyme responsible for the conversion of superoxide radicals into less harmful products like hydrogen peroxide and eliminates secondary toxicity of OH radicals and H2O2 by decreasing the concentration of superoxide radicals (McCord et al.1984). Catalase brings about the reduction of hydrogen peroxides and protects the tissues from the highly reactive hydroxyl radicals (Chance et al.1982). Reduced glutathione (GSH) provides protection to the cells against the toxic effects of lipid peroxidation (Nicotera and Orrenius 1986). GPx uses GSH as a substrate and metabolizes hydrogen peroxide into water (Sies 1993). In the present context too, administration of fluoride in drinking water caused a significant elevation in hepatic and renal tissue lipid peroxidation (51.10, 50.24%) and decreased the levels of TAA (20.37, 24.19%), SOD (35.32, 30.06%), CAT (51.45, 52.31%), GSH (35.77, 29.09%) and GPX (39.72, 33.02%). While basal and HCLP diet fed animal groups did not show any significant variation in hepatic and renal lipid peroxidation and antioxidant levels, fluoride intoxicated rats when fed high protein diet, registered a significant reduction in hepatic and renal lipid peroxidation and increased antioxidant levels (Tables 5 and 6).

Table 5.

Effect of diets on hepatic lipid peroxidation and antioxidant profiles

Groups Control Fluoride control F Basal F HCLP F HPLC
TBARS (mM MDA 100g-1) 10.4 ± 0.22 15.7 ± 0.21a (+51.1) 15.5 ± 0.20a (−1.4) 15.3 ± 0.17a (−2.2) 12.2 ± 0.21ab (−21.8)
Ascorbic acid (μg g-1) 130.4 ± 0.34 103.8 ± 0.28a (−20.3) 104.9 ± 0.20a (+1.05) 105.8 ± 0.35a (+1.8) 121.3 ± 0.43ab (+16.8)
SOD (U mg-1 protein) 4.3 ± 0.20 2.8 ± 0.14a (−35.3) 2.9 ± 0.18a (+2.8) 2.9 ± 0.20a (+5.3) 4.1 ± 0.27b (+47.5)
Catalase (nM H2O2 decomposed Sec-1 g-1) 17.5 ± 0.08 8.5 ± 0.07a (−51.4) 8.6 ± 0.06a (+1.8) 8.5 ± 0.05a (+0.5) 12.1 ± 0.05ab (+42.1)
GSH (mg 100g-1) 41.4 ± 0.27 26.6 ± 0.43a (−35.7) 26.7 ± 0.32a (+0.2) 27.3 ± 0.29a (+2.7) 37.5 ± 0.28ab (+40.8)
GPx (U mg-1 protein) 7.05 ± 0.29 4.2 ± 0.08a (−39.7) 4.3 ± 0.05a (+1.1) 4.3 ± 0.07a (+2.1) 6.1 ± 0.25ab (+43.7)
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Figures in the parenthesis indicate% increase or decrease. Significant at P < 0.05, 0.001; a indicates the comparison with control group and b indicates the comparison with fluoride control group (n=6)

Table 6.

Effect of diets on renal lipid peroxidation and antioxidant profiles

Groups Control Fluoride control F Basal F HCLP F HPLC
TBARS (mM MDA 100g-1) 4.1 ± 0.16 6.2 ± 0.13a (+50.2) 6.2 ± 0.13a (−0.1) 6.2 ± 0.12a (−0.4) 4.8 ± 0.15ab (−22.7)
Ascorbic acid (μg g-1) 69.2 ± 0.28 52.4 ± 0.28 a (−24.1) 53.6 ± 0.39 a (+2.1) 52.5 ± 0.40 a (+0.2) 64.8 ± 0.43 ab (+23.6)
SOD (U mg-1 protein) 3.06 ± 0.15 2.1 ± 0.15 a (−30.06) 2.2 ± 0.10 a (+3.7) 2.1 ± 0.13 a (+0.4) 3.00 ± 0.09 b (+40.1)
Catalase (nM H2O2 decomposed Sec-1 g-1) 6.06 ± 0.04 2.8 ± 0.02 a (−52.3) 2.9 ± 0.02 a (+2.7) 2.9 ± 0.02 a (+1.04) 3.7 ± 0.02 ab (+28.7)
GSH (mg 100g-1) 12.8 ± 0.25 9.1 ± 0.27 a (−29.09) 9.5 ± 0.14 a (+4.05) 9.1 ± 0.26 a (+0.2) 12.8 ± 0.17 b (+40.8)
GPx (U mg-1 protein) 3.2 ± 0.14 2.1 ± 0.06 a (−33.02) 2.3 ± 0.10 a (+6.4) 2.2 ± 0.08 a (+2.3) 2.7 ± 0.11 ab (+26.2)
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Figures in the parenthesis indicate% increase or decrease. Significant at P < 0.05, 0.001; a indicates the comparison with control group and b indicates the comparison with fluoride control group (n=6)

These beneficial effects of the formulated diets could be attributed to the bioactive components of the diets viz., polyphenols, flavonoids, saponins, and ascorbic acid which are known to play important physiological roles in metabolism. They act as antioxidants, antihyperglycaemic and antihyperlipeamic agents, reduce the absorption of cholesterol and increase its excretion (Pandey and Rizvi 2009; Yao et al.2004). The phytochemical analyses of the grains used in the diets indicated the presence of polyphenols, flavonoids, saponins and ascorbic acid content (data not shown) and these grains are also reported to contain phytins and fibers (Gopalan et al.2004).

The marginally beneficial effects of commercial, basal and HCLP diets on one hand and the significant effects of HPLC diet on the other hand could be due to the low protein content in the former compared to the latter. It is perhaps due to the low protein content, all the three diets (commercial, basal and HCLP) could not overcome the fluoride toxicity as indicated by the higher plasma glucose, lipid profiles and lowered antioxidant status with high lipid peroxidation. With a high protein diet, the excretion of fluoride could have been enhanced implying once again the role of proteins in diet as reported earlier (Boyde and Cerklewski 1987; Wang et al.1994; Chinoy and Mehta 1999; Chinoy et al.2006). Since the fluoride inhibition has been removed, the antioxidant profiles of the animals exposed to fluoride improved upon feeding HPLC diet. This improvement in antioxidant activity could be related to the phytoconstituents of the diet as mentioned earlier.

From the forgoing, it becomes clear that both basal and HCLP diets marginally rendered protection from fluoride induced hyperglycemia, hyperlipidemia and oxidative stress. When the protein fraction was increased in the diet it resulted in significant decline in plasma, hepatic carbohydrate and lipid profiles and reduced both hepatic and renal tissue lipid peroxidation. Further, the multigrain diet enriched with protein also improved the antioxidant activity owing perhaps to an increased fluoride excretion. Therefore, a multigrain diet with a high antioxidant potential could be considered a viable option to tackle the fluoride induced toxic effects along with other protein rich foods.

Acknowledgements

The financial assistance in the form of a Research Fellowship to RAV from the University Grants Commission, New Delhi, India is gratefully acknowledged.

Abbreviations

C

Control

FC

fluoride exposed animals

F Basal

fluoride exposed animals given the basal diet

F HCLP

fluoride exposed animals given high carbohydrate low protein diet

F HPLC

fluoride exposed animals given high protein low carbohydrate diet

G-6-Pase

glucose- 6- phosphatase

TL

total lipids

TC

total cholesterol

TG

triglycerides

AI

atherogenic index

TAA

total ascorbic acid

SOD

superoxide dismutase

CAT

catalase

GsH

reduced glutathione

GPx

glutathione peroxidase

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