ABSTRACTCardiovascular fed with apple juice and apple

ABSTRACTCardiovasculardiseases (CVDs) are known as the principal cause of demise and disabilityuniversally and also in Pakistan. In Pakistan heart diseases are the majorreason of death.

CVDs are expanding step by step attributable to poor way oflife and dietary behavior. Hypercholesterolemia is considered as essential factorfor cardiovascular diseases. There are a few sorts of medications that can beutilized for the cure of sicknesses however these medications have diversereactions. From the ancient times many fruits have been considered asbeneficial for health. Apple and Apple cider vinegar have been utilized sincenumerous years for the treatment of various diseases including hypercholesterolemia.The present study aimed to check the combined effect of apple and apple cidervinegar for reducing hypercholesterolemia. Hypercholesterolemia will be inducedin rats by giving them high cholesterol diet. After the induction of hypercholesterolemiathe rats will be fed with apple juice and apple cider vinegar in their diet atthe ratio of 30ml/day and 20 ml/day respectively.

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The blood samples will betaken after 30 days interval. After the treatment cholesterol, triglycerides,LDL and HDL will be estimated from blood serum. Data obtained will be subjectedto an appropriate statistical analysis to determine the level of significance1. INTRODUCTION:CVDs are one of the major metabolicissues that are tormenting our general public at a disturbing rate. It is themain reason of morbidity and mortality around the world. Poor dietary decisionsand way of life are the central point in elevating the disease.

CVDs contributeto 30 percent of all deaths and 80 percent of CVDs related deaths, in lower andmiddle income countries of the world (IOM, 2010). CVDs represent roughly 17.3million deaths per year that is probably increased to more than 23.6 million bythe year 2030. Elevated cholesterol is assessed to cause 2.6 million deaths.

About85.7million, or 34percent, of adults have high blood pressure and about 28.5 million, or 11.9percent, of adult have total cholesterol of 240 mg/dl or higher (AHA, 2017).Hypercholesterolemia is characterizedby high level of cholesterol in blood. Cholesterol is produced by liver and isa waxy substance that caused blockage in blood vessels.

Although cholesterol isessential for different body functions which include synthesis of membrane,production of hormone and fat digestion substances, however a very highcholesterol level increases the risk of heart attack (Benjamin et al., 2010). The main source ofcholesterol is our food and food products such as meat, poultry, fish and eggyolk. These foods consist of saturated and Trans fats that increases thesynthesis of cholesterol from liver and result in hypercholesterolemia (Costa et al., 2003).The fat deposits in the arteries (thatsupply blood to heart) and causes hardening of artery walls that results inatherosclerotic plaques. These plaques act as hurdles between the blood vesseland heart and ultimately increase the risk of heart attack.

There are also somecases in which hypercholesterolemia is inherited disease that cause excesscholesterol to accumulate in body tissues but this case is one in five hundred(Akishita, 2004).            Lipoproteinsare transport protein which are attached to blood and circulate cholesterolwith blood in human body. There are mainly two types of lipoprotein dependingupon the type of cholesterol they carry. Low density lipoprotein (LDL) alsoknown as bad cholesterol carries cholesterol from liver and transport todifferent part of body. In case of its excess cholesterol accumulate in arterywalls and lead to atherosclerosis. High cholesterol lipoprotein (HDL) carriesexcess cholesterol away from liver cells where it breaks down and release as awaste product. That’s why these lipoproteins are known as good cholesterol.

(CDC,2017) .Unhealthy food containing high fat contents, obesity, diabetes, smoking,high blood pressure and physical inactivity increase LDL level and decreasesHDL level which results in stroke and heart failure (Sangsawang et al.,2015). Extensive use of chemical drugs causes problem that issometimes even more hazardous than the disease itself.

As a result, some drugsmay be excluding from consideration or prescribed with extreme caution. Fear ofdrug side effects brings about their improper use and rejection. Therefore, thedisease will not be desirably controlled (Beheshti et al.

, 2012).Because of these issues a large number of studies havebeen performed to check the influence of different products on blood lipids (Shisheehboret al., 2008). It is proved thatapple have many protective effects against cholesterol. Apples also ranked thesecond for total concentration of phenolic compounds, and perhaps moreimportantly, apples had the highest portion of free phenolics when compared toother fruits (Boyer and Lui, 2004). This means that these compounds are notbound to other compounds in the fruits, and the phenolics may be more availablefor eventual absorption into the bloodstream.

            The influence of applecider vinegar has been investigated for hundreds of years. It was in fact firstused about 5000 years ago (Budak et al.,2011). More valuable properties of apple cider vinegar and its ingredients,suggesting their therapeutic effects, have been recently discovered (Fushimi et al., 2006). Apple cider vinegarcontains polyphenolic compounds that have anti oxidant flavonoid which can reducethe deleterious effects of high cholesterol level (Verzelloni et al., 2007).            Apple products arewidely used in world.

A lot of research is available on the use of apple alone,however very little information is available regarding the effect of applecider vinegar towards cholesterol lowering. The main focus of this study is tocheck the effect of apple juice and apple cider vinegar alone and incombination with one another for lowering the cholesterol level (Beheshti et al.,2012).OBJECTIVESThis study will be conducted to achieve thefollowing objectives.·        To check the nutritional profile of apple andapple cider vinegar ·        To evaluate the significance of apple and applecider vinegar in hypercholesterolemic rat 2.

REVIEW OFLITERATUREThere is very close relationshipbetween diet and various chronic diseases. Different fruits and vegetables havepositive effect on chronic diseases due to their phenolic and antioxidantcompounds. Apple and apple cider vinegar is considered as an important fruit inlowering the risk of chronic diseases (Cancer, CVD, Diabetes and Highcholesterol) because it contains chemical component such as flavonoids,polyphenol and carotenoids (Boyer and Lui, 2004). Hyperlipidemia is a result ofaccumulation of triglycerides and cholesterol level in blood.

In Malaysia applecider vinegar is considered as very common home remedy to decrease weight. Astudy in which hypercholesterolemia was induced in rats by giving them high fatdiet conducted by Ajaykumar et al. (2012). 1ml/day apple cider vinegarwas prescribed to rats for 14 days. Cholesterol, high density lipoprotein, lowdensity lipoprotein and very low density lipoprotein was measured. Resultsshowed that there was remarkable decrease in LDL and VLDL and increase in HDLcholesterol.

They negotiate that apple cider vinegar has significantcholesterol lowering ability. Apple cider vinegarhas beneficial effect on lowering the serum level of cholesterol, triglyceridesand LDL after 8 weeks. A study on rats were conducted in which the consequenceof 30 ml apple cider vinegar for 60 days were evaluated and found that therewas adequate decrease in total cholesterol, LDL cholesterol, triglycerides andincreases the HDL cholesterol (Beheshti etal., 2012).Impact ofapples on digestion of lipids on 40 male rats was deliberated. Rats were fedwith 0.3% cholesterol rich food.

In the eating routine of rats, 15% lyophilizedapple was supplemented with glucose, fructose and sucrose. Blood tests weretaken after each 21 days and examined for triglyceride, HDL and cholesterol. Resultsuncovered that slight but significant impact on cholesterol lowering in plasmaand in addition in liver. There was likewise decrease in triglyceride richlipoprotein and increase in HDL part while there was impressive impact incardiovascular protection effect (Aprikian et al., 2002). Effect of whole apple intake on plasma lipids wasobserved.

They concluded that 550g of whole apple intake for 30 dayssignificantly lower the low density lipoprotein cholesterol (Ravn et al.,2013).Another examination was led tocheck the Influence of apples, pears and peaches on lipids on rats .Apples,pears and peaches were fed to rats at the rate of 10% based on dry weight andnon-oxidized cholesterol (NOC) at the rate of 1% was likewise included theeating routine and found that apples and to a less degree pears and peaches supplementeddiet lowers the hypocholesterolemic impact in rats. The high substance ofpolyphenols, phenolic acids and hypolipidemic properties of apples makes thisnatural product best for dietary counteractive action of cardiovascular anddifferent infections (Leontowicz et al., 2002). The effect of dried apple and dried plum (75g/day) on post menopausal women wascompared. Fasting blood samples were collected at baseline, 3, 6 and 12 monthsto measure serum total cholesterol and low density lipoprotein.

The resultsshowed that there was notable reduction in serum level of total cholesterol andhigh density lipoprotein cholesterol in women who consumed dried apple. Thisstudy also concluded that daily apple consumption greatly improved atherogenicrisk factors (Chai et al.,2012). The outcomeof dietary apple polyphenol from unripe apple on rats was determined by givingthem purified diet containing 0.5 % cholesterol and observed that dietary applepolyphenol effectively decreases the low density lipoprotein cholesterol andincreases the high density lipoprotein cholesterol (Osada et al., 2006).

In another study effect of lyophilized apple on plasma andtissue lipids on rats were checked. Experimental diet was equilibrated forsugar supply contained 0.25/100 g cholesterol. Rats were hypercholesterolemicwith cholesterol accumulation in LDL and HDL fractions. Aprikian et al.

(2002) concluded that apple dietlowered plasma and LDL cholesterol (22 and 70 %) and reduced triglycerideaccumulation in heart and liver.Apple juicealso has potential to lower the hypercholesterolemia. Another study wasconducted on rabbits. They categorized the rabbits into four groups,normal diet, high cholesterol diet, 1% cholesterol supplemented with 5 ml applejuice and 1% cholesterol supplemented with 10 ml apple juice for 2 month. Setorki et al. (2009) conclude that10ml apple juice significantly reduced the LDL but 5ml apple juice did not haveany effect on LDL. The effectof apple pomace and apple juice concentrate on fat loss and lowering the cholesterolwas determined.

Rats were fed with diet induced with apple pomace and applejuice at the rate 10% each. After 5 week interval, rats were checked for theirbody weight gain, adipose tissue weight, serum total cholesterol, LDL and HDL.Results of this study showed that there was significant decrease in LDL andincrease in HDL lipoproteins. The results also indicated that both apple pomaceand juice could be helpful for reducing weight as well as improving lipidprofile (Cho et al.

, 2013).  Apple cider vinegar was considered as healthyfruit for the remediation of hyperlipidemia in comparison with drug. In thisstudy they designed different groups in each group 25 healthy subjects werecategorized in the age of 40-60 years with hyperlipidemia. One group wastreated with apple cider vinegar while other group was given drugs. Apple cidervinegar was given as a dose of 5cc two times a day for one month continuously(30 min before lunch and dinner). Lovastatin drug was given at the rate of 20mg/dayfor 4 weeks. Blood samples were taken before and after treatment with regulartime interval. Results of this experiment revealed that there was decrease inthe LDL and cholesterol and increase in HDL level.

There was 25 and 26%decrease in LDL while 35 and 36% increase in HDL level in blood after 4 weeksin case of apple cider vinegar and lovastatin respectively. It is proved thatapple cider vinegar could be used as substitute for drugs with almost sameeffect and with no side effect (Jaferian et al., 2006).

.           Theimpact of apple cider vinegar supplementation on serum triglycerides and totalcholesterol was checked and found that there was remarkable reduction in serumlipid levels (Naziroglu et al.,2014). The combination of apple andapple cider vinegar with other products could also be effective for theremediation of hyperlipidemia. They planned a study to check the combinedeffect of honey and apple cider vinegar for lowering of blood lipids.

Applecider vinegar was given as a dose of 200cc two times a day and honey was givenone tablespoon. Blood lipid levels were measured before and after treatment.Results revealed that there was significant decrease in LDL and increase in HDLby giving apple cider vinegar and honey in combination (Rishehri et al.,2014). MATERIAL AND METHODSArea of ResearchThe research will be conducted in the labs of IHS andNIFSAT, University of Agriculture Faisalabad.Procurement ofRaw MaterialApple and apple cider vinegar will be purchased from localmarket of Faisalabad.Sample PreparationFresh apples will be taken and wash properly with tap water.After drying, peel off the apple.

Slice the apple into small pieces and extractjuice from electric juicer. Apple cider vinegar will be given (Aprikian etal., 2002).

  Proximate AnalysisAnalysis for moisture, ash content, crude protein, and crudefiber, and NFE of apple juice and apple cider vinegar will be carried outaccording to their respective methods (AACC 2000). And this procedure will beconducted in the post graduate research laboratory, National Institute of FoodScience and Technology (NIFSAT), University of Agriculture Faisalabad.Moisture ContentHot air oven will beused to determine the moisture content (AACC, 2000).Crude ProteinTo determine the protein content in sample Kjeldahl’s methodwill be used (AACC,2000).CrudeFat  Soxhlet apparatuswill be used to check the crude fat of sample (AACC, 2000).Crude FiberThe grounded sample will be used for crude fiberdetermination after fat (AACC,2000).Nitrogen Free ExtractNitrogen free extract will also be determined by the methoddescribed in (AACC,2000).

EFFICACY STUDYRats will be purchased from Animal Husbandry DepartmentUniversity of Agriculture Faisalabad  Sample CollectionTwenty rats will be selected randomly and divide into twogroups.Ø  GroupA:  control group                                       (n=5)Ø  GroupB: Experimental group                             (n=15)Where n=number of individuals.Control group will be expressed as T1. Theexperimental group will be further divide into 3 treatment groups as T2,T3, and T4.  Treatment                   Dosage      Apple cider vinegar     Apple juice        T1           Control Control        T2          20ml/day _______         T3         ________ 30ml/day         T4        20ml/day 30ml/day  The apple juice and apple cider vinegar will be added intheir regular diets.

The study will be conducted for 60 days. Initially bloodsample will be taken from each rat and analyze at local laboratory to evaluatethe effect of apple juice and apple cider vinegar on the different biochemicalparameters.Induction of Hypercholesterolemia in RatsHypercholesterolemia will be induced inrats by feeding them on high fat diet (banaspati ghee, corn oil, and cow ghee)for 15 days baseline period (Tembhurne et al., 2012) Biochemical AnalysisBlood samples will be taken from eachrat at 0, 30th and 60th day and analyzed at a locallaboratory to evaluate the effect of apple juice and apple cider vinegar onblood lipid including cholesterol, HDL, LDL, and triglycerides (Dong et al.,2011).  Mineral AnalysisApple juice and apple cider vinegarwill be analyzed for mineral content by wet digestion according to the methodsdescribed in AOAC (2006).StatisticalAnalysisThe data obtained for each parameterwill be analyzed statistically to determine the level of significance(Montgomery, 2008).

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