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EFFECT OF RAW AND PROCESSED OAT BRAN ON LIPID PROFILE OF NORMAL HYPERCHOLESTEROLEMIC AND DIABETIC RATS.

Byline: M. A. Khan F. M. Anjum I. Pasha M. Nadeem and M. A. Sheikh

: ABSTRACT

The core objective of current research is to explore the functional properties of diets containing raw and processed oat bran to combat lifestyle disorders i.e. cardiovascular diseases. For the purpose raw oat bran was procured from Ayub Agricultural Research Institute (AARI) Faisalabad and it was processed through extrusion technique. In addition three types of studies were conducted on the basis of diets i.e. study I (normal rats) study II (hypercholesterolemic rats) and study III (diabetic rats). Moreover the efficacy study was carried out to observe the effects of different levels of raw and processed oat bran on biological profile of blood Sprauge-Dawley rats. Results showed that the cholesterol LDL and triglycerides level reduction was found significantly different when raw and processed oat bran diets fed to normal hypercholesterolemic and diabetic rats. The highest reduction was recorded when fed on diet containing 30% processed oat bran.

The processed oat bran exhibited more reduction as compared to raw oat bran. Conclusively it is suggested that processed oat bran should be introduced in diet based therapy to control lifestyle-related disorders.

Keywords: Oat bran AY-glucan cholesterol LDL triglycerides.

INTRODUCTION

The occurrence of bioactive molecules in the plants provides them imperative position in diet based regimes. However significant amount of these bioactive ingredients have been lost during processing. In addition some of the bioactive compounds hold multifaceted positions for their consumption that these add high nutritional value to the foods (Butt et al. 2007). Cereals especially oat is one example as it contains several nutrients of higher worth and containing phytochemicals. In this perspective presence of bioactive ingredients in oat are of significance importance especially AY-glucan. These ingredients own several operational and nutritional functions.

The AY-glucans in the oat groat are more concentrated in the bran fraction than in the endosperm fraction. Contrary to oats the AY-glucans in barley and rye are more evenly distributed throughout the endosperm (Cui and Wang 2009). Oat bran possess 6.03% moisture 15.23% protein 55.38% carbohydrate 14.13% total dietary fiber (TDF) 4.5% AY-glucans 6.8% fat and 2.43% ash (Usman et al. 2010).

The AY-glucan is a significant constituent of soluble dietary fiber in barley and oats. AY-glucan is generally considered a branched semi-flexible and linear polymer. It is stitched up of glucose units linked by D-(1- 4) with a casual glycoside (1-3)-glycosidic. The configuration of the chain depends on the relative amount of (1-4)- and (1-3)-glycosidic bond between replicating units of glucose. As it is typical polysaccharides in aqueous solutions it has a strong tendency to associate and the viscosities of the solutions are high (Johansson et al. 2000).

The AY-glucan contains both soluble and insoluble components of dietary fiber but oat AY-glucan is classified as soluble fiber (AACC 2000). After over thirty years of extensive research into the numerous and diverse claims for its benefits it is now evident that fiber is distinctive in its considerable physiological benefits (Yun et al. 2003). The AY-glucan is an alternative and appropriate source of dietary fibre that can be added to foods. It can also be added in variety of foods such as dairy cereal meat and other products depends upon the choices for selection of fibre in the diet.

The potential physiological mechanisms behind the efficacy of AY-glucan are its ability to retard the absorption rate of food in the intestine due to increased viscosity thus balancing the cholesterol and triglycerides level in the blood (Wood 2000; Jenkins et al. 2008). Hence keeping in mind the factors discussed above the current study was designed with objective to elucidate the health claims of oat bran AY-glucan.

MATERIALS AND METHODS

Materials: Two oat varieties i.e. Avon and Sargodha-81 grown commercially were procured from Ayub Agricultural Research Institute (AARI) Faisalabad.

Milling: The milling process of oats includes three steps as described below. In the first step cleaning and sizing of the material was carried out to remove the dust chaff rocks other grains and other foreign material from the oats. In the second step dehulling of oat was carried out passing the each variety through de-huller. The outer hull from inner groat of each variety was separated. The lighter oats are separated and groats are taken for further processing. In the final step the groats obtained after dehulling was milled through Quaderumate Senior Mills. The oat bran was separated from flour in several grinding and sieving operations to a coarse fraction (bran) and fine fraction (endosperm flour).

Efficacy studies: The sprague Dawley rats were procured from National Institute of Health (NIH) Islamabad and housed in Animal Room of National Institute of Food Science and Technology UAF. These different efficacy trials were conducted in different studies as described below. The diets were based on the inclusion of dietary fiber which contains AY-glucan as active ingredient.

Study I: Normal rats: The efficacy trials were conducted initially on normal rats. They were acclimatized by feeding basal diet for a period of one week. Then these rats were divided into three groups comprising of 10 rats each in group. The diets prepared from the selected treatments were fed for a period of 8 weeks (Table I II).

The feed and water intake were monitored on daily basis while body weights were recorded weekly throughout the experimental period. Blood samples were collected through cardiac puncture and EDTA coated tubes were used for serum collection (Uchida et al. 2001). Serum samples were further analyzed for various assays by using Microlab-300.

Study II: Induced hypercholesterolemia: The rats were fed on high cholesterol diet containing cholesterol (2%) and cholic acid (0.5%) to induce hypercholesterolemia. After one week lipid profile of each rat was monitored to assess induction of hypercholesterolemia.

Study III: Induced diabetic mellitus: Diabetic mellitus was induced in the rats by injecting Streptozotocin (STZ) @ 65mg/kg body weight dissolved in 0.05M citrate buffer (pH 4.5) intravenously. The blood glucose of each rat was monitored after injecting STZ to check the glucose response. Treatment Plan

Table I: Raw oat bran groups

###Normal rats###Diabetic group###Hypercholestromic group

roups###1###2###3###4###1###2###3###4###1###2###3###4

###D0###D1###D2###D3###D0###D1###D2###D3###D0###D1###D2###D3

Table II: Processed oat bran groups

###Normal rats###Diabetic group###Hypercholestromic group

roups###1###2###3###4###1###2###3###4###1###2###3###4

###D0###D1###D2###D3###D0###D1###D2###D3###D0###D1###D2###D3

Serum lipid profile: The serum lipid profile including cholesterol high density lipoproteins low density lipoproteins triglycerides blood glucose and serum insulin were measured according to their respective protocols. The total cholesterol concentration was estimated by liquid cholesterol CHOD-POP method described by Allain (1974). High-density lipoprotein concentration was estimated by using the HDL- cholesterol kits by the method described by Assmann (1979). The serum samples were also analyzed for low density lipoproteins (LDL) following the procedure of McNamara et al. (1990). The blood triglycerides concentration was estimated by liquid triglycerides GPO- PAP method described by Anoni et al. (1982). Statistical Analysis: The data obtained for each parameter were subjected to analysis of variance using Statistical Package (Costat-2003 Co-Hort v 6.1.) through 2-factor factorial CRD. Each treatment contains three replicates.The levels of significance (P=0.05 and P=0.01) following the principles outlined by Steel et al. (1997).

RESULTS AND DISCUSSION

Effect of oat bran on lipid profile of normal hypercholestrolemic and diabeteic rats: In normal rats cholesterol level ranged from 84.74 to 81.53 mg/dL and 85.03 to 81.56 mg/dL when fed on different diets containing various levels of raw and processed oat bran respectively (Table III). Likewise in hypercholestrolemic rats cholesterol level decreased from 226.84 to 203.75 mg/dL and 225.56 to 198.22 mg/dL after the administration of diets containing raw and processed oat brans respectively. However in diabeteic rats 1.13% 4.34% and 6.77% reduction in cholesterol was observed in case of raw oat bran whilst 3.36% 6.49% and 9.13% reduction in cholesterol was noticed with 10 20 and 30% processed oat bran diets (Table V).

The normal rats when given diets containing 10 20 and 30% raw oat bran showed 2.13 3.38 and 4.38% reduction in serum LDL respectively as compared to normal rats fed on control diet. In case of diets containing 10 20 and 30% processed oat bran resulted 2.54 3.45 and 5.23% decrease in serum LDL respectively (Table VI). Among the hypercholesterolemic rats the highest LDL level was recorded in the rats fed on control diet and it reduced from 145.84 to 124.73 mg/dL and 151.53 to 126.03 mg/dL fed on diets containing various levels of raw and processed oat bran respectively (Table VII). In diabetic rats the diet containing 30% processed oat bran showed the highest reduction (11.92%) in LDL but at same level of raw oat bran this reduction was 9.42% (Table VIII). Regarding HDL level of normal rats fed on raw and processed oat bran the results in Table IX indicated that it ranged from 38.57 to 39.40 mg/dL and 38.49 39 to 40.17 mg/dl respectively. In hypercholesterolemic rats the diets containing 10 20 and 30% raw oat bran showed 1.70 3.66 and 5.76% increase in HDL level respectively while the diets containing 10 20 and 30% processed oat bran exhibited 2.19 4.40 and 7.10% increase in HDL level respectively (Table X). However the HDL level in diabetic rats ranged from 39.15 to 41.08 mg/dL in raw oat bran diets and 39.56 to 41.83 mg/dL in processed oat bran diets (Table XI).

The triglycerides level in normal rats decreased from 74.20 to 72.89 mg/dL when fed on raw oat bran and 74.44 to 71.87 mg/dL in processed oat bran diets (Table XII). But In hypercholesterolemic rats the triglycerides level varied from 106.54 to 101.33 mg/dL fed on raw oat bran whilst showed reduction from 106.50 to 98.67 mg/dL when fed on processed oat bran (Table XIII). However the triglycerides level was found significantly the highest in the diabetic rats when fed on control diet and it varied from 95.59 to 91.92 mg/dL and 96.90 to 92.45 mg/dL when different levels of raw and processed oat bran was given respectively (Table XIV).

Table III. Effect of raw and processed oat bran diets on cholesterol (mg/dL) of normal rats

###ROB (mg/dL)###POB(mg/dL)

###2010###2011###Mean###2010###2011###Mean

Control###84.73###84.74###84.74a###85.48###84.58###85.03a

10%###83.62###83.76###83.69b###84.00###83.13###83.57b

20%###83.19###82.65###82.92c###83.17###82.35###82.76bc

30%###81.55###81.51###81.53d###81.96###81.17###81.56c

Mean###83.27###83.17###83.65###82.81

Table IV. Effect of raw and processed oat bran diets on cholesterol (mg/dL) of hypercholesterolamic rats.

###ROB (mg/dL)###POB(mg/dL)

###Diets

###2010###2011###Mean###2010###2011###Mean

###Control###223.18###230.50###226.84a###223.43###227.69###225.56a

###10%###209.10###214.56###211.83b###207.87###210.25###209.06b

###20%###204.45###210.20###207.33ab###209.66###200.26###204.96b

###30%###200.39###207.11###203.75b###195.90###200.54###198.22c

###Mean###209.28 aa###215.59###209.22###209.69

Table V. Effect of raw and processed oat bran diets on cholesterol (mg/dL) of diabetic rats

###ROB (mg/dL)###POB(mg/dL)

###Diets

###2010###2011###Mean###2010###2011###Mean

###Control###116.42###117.40###116.91a###119.00###117.83###118.41a

###10%###115.12###116.06###115.59b###115.88###112.97###114.43b

###20%###111.34###112.34###111.84c###112.49###108.95###110.72c

###30%###109.84###108.16###109.00d###108.03###107.17###107.60c

###Mean###113.18###113.49###113.85###111.73

Table VI. Effect of raw and processed oat bran diets on LDL (mg/dL) of normal rats

###ROB (mg/dL)###POB(mg/dL)

###Diets

###2010###2011###Mean###2010###2011###Mean

###Control###30.62###30.30###30.46a###30.70###30.81###30.76a

###10%###29.95###29.67###29.81ab###30.08###29.88###29.98b

###20%###29.72###29.14###29.43bc###29.79###29.60###29.70b

###30%###29.09###28.88###28.99c###29.26###29.05###29.15c

###Mean###29.85###29.50###29.96###29.84

Table VII. Effect of raw and processed oat bran diets on LDL (mg/dL) of hypercholesterolamic rats

###ROB (mg/dL)###POB(mg/dL)

###Diets

###2010###2011###Mean###2010###2011###Mean

Control###141.87###149.82###145.84a###151.58###151.47###151.53a

10%###133.88###144.51###139.20ab###142.38###141.22###141.80b

20%###127.64###135.19###131.41cd###136.67###135.15###135.91c

30%###121.36###128.09###124.73c###125.63###126.43###126.03d

Mean###131.19###139.40###139.07###138.57

Table VIII. Effect of raw and processed oat bran diets on LDL (mg/dL) of diabetic rats

###ROB (mg/dL)###POB(mg/dL)

###Diets

###2010###2011###Mean###2010###2011###Mean

Control###58.52###57.14###57.83a###57.55###56.25###56.90a

10%###57.23###55.71###56.47b###55.46###54.36###54.91b

20%###56.78###55.12###55.95b###53.21###53.29###53.25c

30%###51.98###52.78###52.38c###49.35###50.89###50.12d

Mean###56.13###55.19###53.89###53.70

Table IX. Effect of raw and processed oat bran diets on HDL (mg/dL) of normal rats

###ROB (mg/dL)###POB(mg/dL)

###Diets

###2010###2011###Mean###2010###2011###Mean

Control###38.57###38.57###38.57###38.14###38.84###38.49

10%###38.90###38.75###38.86###38.72###39.10###38.91

20%###39.36###38.91###39.14###39.55###39.37###39.46

30%###39.71###39.09###39.40###40.76###39.58###40.17

Mean###39.15###38.83###39.29###39.22

Table X. Effect of raw and processed oat bran diets on HDL (mg/dL) of hypercholesterolamic rats

###ROB (mg/dL)###POB(mg/dL)

###Diets

###2010###2011###Mean###2010###2011###Mean

Control###53.40###57.00###55.20c###56.42###57.06###56.74b

10%###55.63###56.65###56.14bc###58.27###57.68###57.98ab

20%###59.29###55.32###57.30ab###60.62###58.08###59.35a

30%###61.07###55.69###58.38a###61.70###59.84###60.77a

Mean###57.35###56.17###59.25###58.16

Table XI. Effect of raw and processed oat bran diets on HDL (mg/dL) of diabetic rats

###ROB (mg/dL)###POB(mg/dL)

###Diets

###2010###2011###Mean###2010###2011###Mean

Control###38.83###39.47###39.15c###39.15###39.97###39.56b

10%###39.20###40.22###39.71b###40.25###40.72###40.49ab

20%###41.52###39.15###40.34ab###42.05###39.67###40.86ab

30%###41.77###40.79###41.08a###42.64###40.78###41.83a

Mean###40.33###39.90###41.02###40.28

Table XII. Effect of raw and processed oat bran diets on triglycerides (mg/dL) of normal rats

###ROB (mg/dL)###POB(mg/dL)

###Diets

###2010###2011###Mean###2010###2011###Mean

Control###74.03###74.37###74.20###74.84###74.03###74.44

10%###74.17###73.84###74.01###73.82###73.94###73.88

20%###75.01###74.00###73.50###73.84###72.82###73.33

30%###72.79###72.99###72.89###72.02###71.73###71.87

Mean###73.50###73.80###73.63###73.13

Table XIII. Effect of raw and processed oat bran diets on triglycrides (mg/dL) of hypercholesterolamic rats

###ROB (mg/dL)###POB(mg/dL)

###Diets

###2010###2011###Mean###2010###2011###Mean

Control###106.54###106.55###106.54a###106.79###106.21###106.50a

10%###103.45###105.45###104.45ab###99.98###107.99###103.99ab

20%###106.12###100.30###103.21b###102.40###102.14###102.27bc

30%###102.33###100.33###101.33b###98.50###98.84###98.67c

Mean###104.61###103.16###101.92###103.80

Table XIV. Effect of raw and processed oat bran diets on triglycerides (mg/dL) of diabetic rats

###ROB (mg/dL)###POB(mg/dL)

###Diet

###2010###2011###Mean###2010###2011###Mean

Control###95.40###95.77###95.59a###97.49###96.31###96.90a

10%###93.12###95.38###94.25b###95.44###95.03###95.24b

20%###92.92###94.10###93.51b###94.20###94.07###94.14c

30%###92.30###91.54###91.92c###92.50###92.40###92.45d

Mean###93.44###94.20###94.91a###94.45

It is evident from the results regarding lipid lowering attribute of raw and processed oat bran from different studies i.e. normal hypercholesterolemic and diabetic rats showed that the addition of either raw or processed oat bran in the diets significantly reduced cholesterol LDL and triglycerides and enhance HDL level in all studies however processed oat bran exhibited better results. Overall it was observed that bran supplementation in any form proved more helpful to attenuate the lipid abnormalities.

In the present studies maximum decline in cholesterol level in rats fed on diets containing processed oat bran was observed than the rats fed on diets containing raw oat bran which might be due to differences in the extractability of AY-glucan regarding raw and processed oat bran. The processing methods have increased the extractability of AY-glucan and it ultimately increased the AY-glucan content in processed diets of rats so it reduced more cholesterol level than rats fed on raw oat bran diet.

The findings of Andersson et al. (2010) have supported the results found in the present study in which they observed reduction in cholesterol LDL and triglycerides in mice fed on diets containing raw or processed oat bran. The results of some other studies are also in agreement with the findings of present studies which showed a decline trend in plasma lipid profile especially cholesterol with raw and processed oat bran diets ( Welch et al. 1995). Immerstrand et al. (2009) have also reported that hypercholesterolemic rats fed on processed oat bran diet exhibited significant reduction (17%) in plasma cholesterol. In another study Anderson et al. (2000) reported a decline in plasma cholesterol of rats fed on oat bran AY-glucan. Likewise results have been found in hypercholesterolemic rats on cholesterol lowering when subjected to raw or processed oat bran diets.

The present results showing significant effect of diets containing oat bran on HDL are in agreement with previous work of Kerkhof et al. (2003) who reported considerable changes for HDL in human subjects after consumption of oat bran bread. Similarly Beck et al. (2009) reported changes in HDL of human subjects by daily intake of oat AY-glucan. The findings of Deveries (2004) also supported the present results as they stated that HDL-cholesterol increased after consumption of a diet containing oat bran. The results for momentous effects of diet containing oat bran are in line with Andersson et al. (2000) investigation they expounded that oat bran AY-glucan affected HDL significantly.

The triglycerides are fats that can clog arteries and increase risk for heart attacks and stroke. The dietary fiber especially oat bran has shown to reduce triglycerides level. The reduction in plasma triglyceride has been reported due to a decrease in hepatic secretion of low density lipoproteins (LDL) and very low density lipoproteins (VLDL) determined after blocking the clearance of triglyceride-rich lipoproteins with soluble fibers that is a characteristic of all cholesterol-lowering fibers.

The oat bran AY-glucan also involves in the bile acid circulation and it binds the bile acids and decreases their absorption from the intestine. As a result LDL cholesterol is removed from the blood and converted the bile acids by the liver to replenish the bile acids excreted in the stool and ultimately reduced the triglycerides level in the blood. The same mechanism for regulating triglycerides by oat bran supported the present results of decreasing triglycerides by oat bran in Sprague Dawley rats have been reported by several researchers (Kerkhoffs et al. 2003).

Conclusion: It is evident that diets containing processed oat bran appeared more effective in modulating lipid metabolism as compared to diet containing raw oat bran. In addition oat bran is a good source of dietary fiber with special reference to soluble AY-glucans. The highest cholesterol reduction was recorded when fed on diet containing 30% processed oat bran. However percentage of decreasing cholesterol was recorded in an ascending order i.e hypercholesterolemic greater than diabetic greater than normal rats. The presence of active ingredients especially AY-glucan could be beneficial in managing the lifestyle related dieases.

Acknowledgements: The authors are highly thankful to Higher Education Commision (HEC) Pakistan for providing funds to accomplish the current study.

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