Kari Sambaiah

Effect of dietary ghee—the anhydrous milk fat, on blood and liver lipids in rats

Dairy products are important sources of dietary fat in India. Anhydrous milk fat, viz., ghee, is consumed as such in the diet and also is used for frying the dishes. Ghee contains high levels of saturated fatty acids and cholesterol, which are considered risk factors for cardiovascular diseases. In the present study, ghee, at levels ranging from 0.25 to 10%, was included in a nutritionally balanced AIN-76 diet fed to Wistar rats for a period of 8 weeks. The serum lipid profiles of these animals showed a dose dependent decrease in total cholesterol, low density lipoproteins and very low density lipoproteins cholesterol, and triglyceride levels when ghee was present at levels greater than 2.5% in the diet. Liver cholesterol and triglycerides also were decreased in these animals. When ghee was included as a sole source of fat at a 10% level, polyunsaturated fatty acids in the serum and liver lipids were reduced significantly. Similar results were observed when ghee was subjected to a higher temperature (120 degrees C) to generate cholesterol oxidation products and fed to the animals. Although cholesterol oxidation products were not accumulated in serum, significant amounts were accumulated in liver only when ghee was fed as a sole source of fat at a 10% level. This study revealed that the consumption of ghee up to a 10% level in the diet altered blood lipid profiles in such a manner as not to elevate the risk factors for cardiovascular diseases.

Dietary garlic and onion reduce the incidence of atherogenic diet-induced cholesterol gallstones in experimental mice

Mice fed with diet containing 0.5 % cholesterol for 10 weeks resulted in cholesterol supersaturation in gallbladder bile which promoted the formation of cholesterol gallstones (CGS). In this study, dietary hypocholesterolaemic spices, garlic and onion (both raw or heat-processed) were examined for their antilithogenic potential by including at 0.6 and 2.0 % level, respectively, along with lithogenic (LG) diet for 10 weeks. Dietary garlic and onion reduced the CGS incidence by 15-39 %, the effect being maximum in the heat-processed onion group. Dietary garlic and onion markedly reduced biliary cholesterol. The cholesterol:phospholipid ratio which was 1.58 in the LG diet group was reduced to 0.73-0.96 in the garlic and onion groups. The biliary cholesterol saturation index was 0.92, 1.25, 1.09 and 0.86, respectively, in the heat-processed onion, raw garlic, heat-processed garlic and raw onion groups, while it was 1.9 in the LG group. The hydrophobicity index of bile was - 0.08, - 0.079, - 0.032 and - 0.073, respectively, in the heat-processed onion, raw garlic, heat-processed garlic and raw onion groups, while it was +0.054 in the LG group. Hepatic hydroxymethyl glutaryl-CoA reductase activity was lowered in the LG diet-fed group, while dietary garlic or onion countered this alteration and also increased the activities of hepatic cholesterol 7 alpha-hydroxylase and sterol 27-hydroxylase. Serum and liver cholesterol were decreased by feeding garlic or onion compared to the LG diet. Thus, dietary Allium spices exerted antilithogenic influence by decreasing the cholesterol hyper-secretion into bile and increasing the bile acid output thus decreasing the formation of lithogenic bile in experimental mice.

Dietary fibres ameliorate decreased synthesis of heparan sulphate in streptozotocin induced diabetic rats

The role of dietary fibers in diabetes has been studied by several workers. Long term dietary treatment with increased amounts of fiber-rich low-glycaemic index natural foods improves blood glucose and reduces the number of hypoglycemic events in type I diabetic patients. On the other hand Rohrbach and Martin and Cohen and Surma described changes in the general and biochemical structure of renal tissues such as the glomerular basement membranes. One of these changes was the reduction and undersulfation of the glycoconjugate and glycosaminoglycan heparan sulfate, which plays an important role in renal structure and function. The purpose of the present study was to determine specific effects of two types of dietary fiber on the composition of kidney glycoconjugates in an animal model of diabetes type I. Streptozotocin-treated diabetic rats were fed either a control diet or diets containing 10% wheat bran (insoluble dietary fiber) or 5% guar gum (soluble dietary fiber). Effects of these fibers on glycaemic control and nephropathy were assessed using previously described methodologies. The effect of dietary fiber in the glycoconjugate composition of kidneys of control and diabetic animals was studied by estimating their total hexose content, sulfated glycosaminoglycans, hexosamines and uronic acids. The activities of enzymes that participate in the synthesis of saccharides and glycoconjugates (L-glutamine-fructose-6-phosphate aminotransferase) and their degradation (N-acetyl-beta-glucosaminidase and beta-glucuronidase) were also evaluated. Results indicated that both soluble and insoluble dietary fibers ameliorated a significant increase in the activity of GFAT. Heparan sulfate was also isolated and quantified. Results indicated that the renal content of heparan sulfate decreased in diabetic animals and that this decrement was ameliorated by the ingestion of both soluble and insoluble fiber in the diet.

Regression of preestablished cholesterol gallstones by dietary garlic and onion in experimental mice.

We have recently reported the health beneficial potential of dietary garlic and onion in reducing the incidence and severity of cholesterol gallstone (CGS) during its experimental induction in mice. In the current study, the efficacy of dietary garlic and onion in regressing preestablished CGS was investigated in experimental mice. After inducing CGS in mice with a lithogenic diet for 10 weeks, they were maintained on basal diets containing 0.6% dehydrated garlic or 2% dehydrated onion for a further 10 weeks. Dietary garlic and onion, either raw or heat processed, regressed preformed CGS in mice up to 53% to 59%, whereas the regression in the basal control diet group was only 10%. The antilithogenic potency of garlic was decreased by its heat processing, but not in the case of onion. Biliary cholesterol was significantly decreased in garlic- and onion-fed animals. Biliary cholesterol saturation index and hydrophobicity index were significantly lowered by dietary garlic and onion. Serum and liver cholesterol levels were decreased by feeding these spices during post-CGS induction period. Hepatic hydroxymethylglutaryl-coenzyme A reductase activity was increased after feeding garlic and onion, whereas activities of the cholesterol-degrading enzymes cholesterol-7α-hydroxylase and sterol-27-hydroxylase were increased in spice-fed groups. These results indicate that feeding garlic and onion effectively accelerates the regression of preformed CGS by promoting cholesterol desaturation in bile. This observation is significant in the context of evolving dietary intervention strategy to address regression of existing CGS and stopping the possible recurrence.

Spray-dried milk supplemented with α-linolenic acid or eicosapentaenoic acid and docosahexaenoic acid decreases HMG Co A reductase activity and increases biliary secretion of lipids in rats

In our earlier study, we have shown that rats fed spray-dried milk containing alpha-linolenic acid (LNA 18:3 n-3) or eicosapentaenoic acid (EPA 20:5 n-3) and docosahexaenoic acid (DHA 22:6 n-3) had significantly lower amounts of serum and liver cholesterol. To evaluate the mechanism for hypocholesterolemic effect of n-3 fatty acids containing milk formulation, we fed male Wistar rats with spray-dried milk containing linseed oil (LSO) (source of LNA) or fish oil (FO) (source of EPA+DHA) for 8 weeks. Feeding n-3 fatty acid containing milk formulation lowered the hepatic 3-hydroxy-methylglutaryl coenzyme A (HMG Co A) activity by 17-22% compared to rats given control diet devoid of n-3 fatty acids. The cholesterol level in liver microsomes was found to be decreased by 16% and 20%, respectively, in LSO and FO containing formulation fed rats. The bile flow was enhanced to an extent of 19-23% in experimental groups compared to control animals. The biliary cholesterol and phospholipid secretion was increased to an extent of 49-55% and 140-146%, respectively, in rats fed n-3 fatty acid containing formulation. The increase in the total bile acids secretion in bile was mainly reflected on an increase in the levels of taurine conjugated bile acids. These results indicated that n-3 fatty acid containing spray-dried milk formulation would bring about the hypocholesterolemic effect by lowering HMG Co A reductase activity in liver and by increasing the secretion of bile constituents.

Modulatory effect of butyric acid-a product of dietary fiber fermentation in experimentally induced diabetic rats.

The effect of feeding of butyric acid on alleviation of diabetic status was studied. Diabetes was induced in rats using streptozotocin. Rats were fed with basal diet containing wheat bran (5%) as a source of insoluble dietary fiber and guar gum (2.5%) as a source of soluble dietary fiber. The experimental group received butyric acid at 250, 500 and 750 mg/kg body weight/day. The diabetic animals lost weight in spite of high diet consumption. The levels of water intake, urine output, urine sugar, fasting blood sugar increased during diabetic condition compared to control and these were reduced by nearly 20% in the fiber-fed diabetic group. Further supplementation of butyric acid at 500 mg/kg body weight/day ameliorated the diabetic status by nearly 40%. Urine sugar level during the diabetic state was reduced from 7.2 g/day to 3.6 g/day and fasting blood glucose from 270 mg/dl to 180 mg/dl. Butyric acid feeding at 500 mg/kg body weight/day was most effective in controlling the diabetic status.

Effect of dietary fibre on intestinal and renal disaccharidases in diabetic rats

Diabetes was induced in rats with streptozotocin. They were fed either with wheat bran as a source of insoluble dietary fibre or guar gum as soluble dietary fibre. The effect of these fibres on intestinal and renal disaccharidases in diabetic rats was studied. Activities of intestinal maltase, sucrase and lactase increased during diabetes, which was ameliorated to a considerable extent in fibre fed groups. The activities of disaccharidases in renal tissue was decreased in diabetic rats and there was an improvement in fibre fed group.

Hypocholesterolemic effect of anhydrous milk fat ghee is mediated by increasing the secretion of biliary lipids.

The anhydrous milk fat ghee is one of the important sources of fat in the Indian diet. Our earlier studies showed that rats fed diets containing greater than 2.5 wt% of ghee had lower levels of serum cholesterol compared with rats fed diets containing groundnut oil. To evaluate the mechanism of the hypocholesterolemic effect of ghee, male Wistar rats were fed a diet containing 2.5 or 5.0 wt% ghee for a period of 8 weeks. The diets were made isocaloric with groundnut oil. Both native and ghee heated at 120 degrees C containing oxidized lipids were included in the diet. The ghee in the diet did not affect the 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase activity in the liver microsomes, but it significantly increased biliary excretion of cholesterol, bile acids, uronic acid, and phospholipids. The rats fed ghee had lower levels of cholesterol esters in the serum as well as in the intestinal mucosa. Both native and oxidized ghee influenced cholesterol metabolism. These results indicate that supplementation of diets with ghee lipids would increase the excretion of bile constituents and lower serum cholesterol levels.

Supplementation and delivery of n-3 fatty acids through spray-dried milk reduce serum and liver lipids in rats.

Indian diets comprising staples such as cereals, millets, and pulses provide 4.8 energy % from linoleic acid (18∶2n−6) but fail to deliver adequate amounts of n−3 FA. Consumption of long-chain n−3 PUFA such as EPA (20∶5n−3) and DHA (22∶6n−3) is restricted to those who consume fish. The majority of the Indian population, however, are vegetarians needing additional dietary sources of n−3 PUFA. The present work was designed to use n−3 FA-enriched spray-dired milk powder to provide n−3 FA. Whole milk was supplemented with linseed oil to provide α-linolenic acid (LNA, 18∶3n−3), with fish oil to provide EPA and DHA, or with groundnut oil (GNO), which is devoid of n−3 PUFA, and then spray-dired. Male Wistar rats were fed the spray-dired milk formulations for 60 d. The rats given formulations containing n−3 FA showed significant increases (P<0.001) in the levels of LNA or EPA/DHA in the serum and in tissue as compared with those fed the GNO control formulation. Rats fed formulations containing n−3 FA had 30–35% lower levels of serum total cholesterol and 25–30% lower levels of serum TAG than control animals. Total cholesterol and TAG in the livers of rats fed the formulations containing n−3 FA were lower by 18–30% and 11–18%, respectively, compared with control animals. This study showed that spray-dried milk formulations supplemented with n−3 FA are an effective means of improving dietary n−3 FA intake, which may decrease the risk factors associated with cardiovascular disease.

Butyric acid modulates activities of intestinal and renal disaccharidases in experimentally induced diabetic rats.

To elucidate the effect of feeding of butyric acid on disaccharidase activities, the specific activities of the disaccharidases were measured in the intestinal mucosa and kidney cortex of control and diabetic rats. Diabetes was induced in rats using streptozotocin. Rats were fed with basal diet containing wheat bran (5%) as a source of insoluble dietary fiber and guar gum (2.5%) as a source of soluble dietary fiber. The experimental group received butyric acid at 250, 500 and 750 mg/kg body weight/day. Increased activities of intestinal maltase, sucrase and lactase in diabetic rats were significantly reduced in fiber-fed diabetic group. Supplementation of butyric acid at 500 mg/kg body weight/day showed a further decrease in their activities. The activity of disaccharidases in renal tissue was decreased in diabetic rats and was significantly improved in fiber-fed diabetic group. Butyric acid feeding at 500 mg/kg body weight/day showed further improvement in their activities.