Research Article
Effect of Jerusalem Artichoke and Olive Oil on Hyperglycemic Rats
Shimaa M Aboelnaga and Fahmida kahtoon*
Corresponding Author: Fahmida Khatoon, Department of Biochemistry, College of Medicine, University of Ha’il
Received: August 14, 2020; Accepted: September 07, 2020 Available Online: September 29, 2020
Citation: Aboelnaga SM & Khatoon E. (2020) Effect of Jerusalem Artichoke and Olive Oil on Hyperglycemic Rats. J Genet Cell Biol, 4(1): 207-216.
Copyrights: ©2020 Aboelnaga SM & Khatoon E. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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The aim of this study was to investigate the effect of two levels of jerusalem artichoke (Helianthus tuberosus L.), two dosage of olive oil and their combination on rats suffering from diabetes. Forty-eight male albino rats (Sprague Dawley Strain) utilized in this work, the rats study divided into two essential groups. The first (n=6 rats) fed on standard diet and used as a control negative group. The second (42 rats) treated with alloxan (150 mg/kg b.wt.) to induce diabetes. The rats in the second main group were divided into 7 subgroups. Subgroup (1): was fed on standard diet and used as a positive control group. Subgroups (2&3): were fed diet containing (2% and 4% Helianthus tuberosus), respectively. Subgroup (4 & 5): were fed standard diet and treated daily with (0.25 ml and 0.5 ml olive oil/rat), respectively. Subgroup (6): was fed diet containing 2% dried Helianthus tuberosus powdered and treated daily with 0.25 ml olive oil/each rat. Subgroup (7): was fed diet containing 4% dried Helianthus tuberosus powdered and treated with 0.5 ml olive oil/each rat daily. This experiment lasted 4 weeks. Results showed that, (liver and kidney weight/body weight%), glucose, lipid fractions except HDL-cholesterol, liver parameters and kidney functions increased significantly in the positive control group, while weight of rats decreased. Treating rats, which were, suffering from diabetes with two level from Helianthus tuberosus (2% and 4%), two dosages from olive oil (0.25ml and 0.5ml olive oil/each rat) and the same dosage from olive oil with the same levels from Helianthus tuberosus together improved all above parameters. Group of rats which fed on diet containing (4% Helianthus tuberosus and treated with 0.5 ml olive oil/each rat) recorded the best results, followed by the groups which were treated with (2% Helianthus tuberosus and 0.25 ml olive oil/each rat) and 4% Helianthus tuberosus, respectively. From our results, it could be concluded that, Jerusalem Artichoke "and olive oil (alone or together) decrease the side effects induced from diabetes.

Keywords: Diabetes - Helianthus tuberosus L, Olive oil, Glucose, Liver enzymes, lipid profile, kidney function
INTRODUCTION

Diabetes (DM) is a complex and varied group of disorders that are a major source of morbidity in the world [1]. DM is defined as a condition in which the balance of carbohydrates and lipid metabolism is incorrectly controlled by insulin. Two major forms in DM: insulin dependent diabetes mellitus (Type 1) and non-insulin dependent diabetes mellitus (Type 2) [2].

Danaei et al. [3] reported that, more than 346 million of individuals around the world suffer from this disease. The lifestyle, intake of high energy and obesity are inducing the rise of diabetes [4]. The incidence of diabetes will increase by the year 2030 to reach 7.7% of the total adult population [5]. On the other hand, diabetes and its related complications still present a major medical problem, the decrease in pancreatic βcell mass will eventually lead to chronic hyperglycemia [6].

Olive oil has traditionally been the principal oil of the Mediterranean diet. The monounsaturated fatty acid in the diet prevents the accumulation of body fat and decreases postprandial adiponectin which gets induced by a carbohydrate rich diet in subjects with insulin-resistant in addition to, polyphenols present in olive oil, have an important antioxidant and anti-inflammatory effect [7,8].

Helianthus tuberosus (HT) tubers contains inulin also a non-degradable oligosaccharide polymer that includes 30-35 fructose units that are linearly arranged in β-1,2 linkages with an α-1,2-linked D-glucose at the non-reducing end [9]. The Jerusalem artichoke tubers (Helianthus tuberosus L.) are known to be a health-promoting source. Jerusalem artichoke contains an important homopolysaccharide (Inulin) which play very important roles for nutrition and treatment of diabetic patients and obesity [10]. On the other hand, Duan et al. [11] indicated that inulin improved glucose tolerance and liver lipid profile in rats fed a high-fat diet (HFD).  

Therefore, the present study was carried out to assess the effects of jerusalem artichoke (Helianthus tuberosus L.), olive oil and their combination on rats suffering from diabetes.

MATERIALS & METHODS

Materials
- Casein, vitamins, minerals, cellulose, alloxan and choline chloride were obtained from El-Gomhoria Company, Cairo Egypt.
- Corn starch, soybean oil, sucrose and olive oil were purchased from local market, Cairo, Egypt.
- Jerusalem artichoke tubers (Helianthus tuberosus L.) was obtained from Agricultural Research Center, Giza Egypt.
- Forty-Eight male albino rats (Sprague Dawley Strain), their weights (150 ± 10 g) were obtained from experimental animal farm, Helwan, Cairo, Egypt.

METHODS

Dried Jerusalem artichoke tubers: Jerusalem artichoke tubers (Helianthus tuberosus L.) were washed thoroughly with water and dried with solar energy in ovens for 24 h, at 40-50˚C. The dried samples were finely powdered by using grinder and stored in bags in deep freezer, until used.

Chemical Analysis of Helianthus tuberosus and fatty acid composition of olive oil: Moisture, protein, ash, crud fiber, oil and soluble carbohydrates were determined in Helianthus tuberosus [12], while inulin was determined by another method [13].

Fatty acid compositions of olive oil were determined according to [14,15].

Biological Part: Forty-Eight male albino rats (Sprague Dawley Strain) their weights (150 ± 10 g) were kept in individual cages under hygienic conditions and fed on basal diet for one week for adaptation period, according to [16]. After this period, the rats divided into two main groups. The first main group (6 rats) fed on basal diet and used, as a control negative group (healthy group). The second main group (42 rats) injected with 150 mg alloxan/kg body weight to cause diabetes, according to another method [17]. Blood samples were collected from the eye of all rats after five days in order to determine the levels of glucose to insure the induction of diabetes. The rats in the second main group were divided into seven subgroups (n = 6) according to the following:

Subgroup (1): Fed basal diet, as a control positive group (diabetic group). Subgroup (2 and 3): were fed basal diet containing (2% and 4% Helianthus tuberosus), respectively. Subgroup (4 and 5): were fed basal diet and treated daily with (0.0.25 ml and 0.5 ml olive oil/rat), respectively. Subgroup (6): was fed basal diet containing 2% dried Helianthus tuberosus powdered and treated daily with 0.25 ml olive oil. Subgroup (7): was fed basal diet containing 4% dried Helianthus tuberosus powdered and treated daily with 0.5 ml olive oil.

During the experimental period (4 weeks), the diets consumed, and body weights were recorded every week. At the end of the experimental period, all rats were fasted overnight, and sacrificed after being anaesthetized. Blood samples were collected from the aorta of all rats (negative, positive and treated groups). Blood samples were centrifuged and then, serum was separated to estimate some biochemical parameters, i.e. serum (glucose, cholesterol, triglycerides, high density lipoprotein HDL-c," low & very low-density lipoprotein-c", uric acid, urea nitrogen, creatinine, “Aspartate Amine Transaminase & Alanine Amine Transaminase” and Alkaline Phosphatase) according to other methods [18-27], respectively.

Liver and kidney in each rat were separated and weighted to estimate liver and kidney weights/body weights %. The obtained results were statistically analyzed (mean ± standard deviation and using one-way ANOVA test) in order to identify the significant differences, by using SAS package (least significant differences at P< 0.05) [28].

RESULTS AND DISCUSSION

Data in this research showed the chemical composition of Helianthus tuberosus L., the nutrients content of Helianthus tuberosus L. presented in Table 1. The amounts of moisture, protein, oil, fiber, ash and soluble carbohydrates were 9.730%, 8.920%, 0.718%, 3.492%, 6.695% and 70.445%, respectively. The content of inulin in Helianthus tuberosus was 23.531%.
Fatty acid composition of olive oil: The fatty acids composition of olive oil was determined by gas liquid chromatography (GLC). The contents of fatty acid in olive oil presented in Table 2. Saturated fatty acids “SFA” in olive oil recorded 18.722%. Palmitic acid (C16:0) was the main saturated fatty acid “15.95%”, while the lowest percent of saturated fatty acid in this oil recorded for stearic acid (C18:0) “2.772%”. Monounsaturated fatty acids “MUFA” in olive oil “64.178%”, the percent oleic acid (C18:1) recorded the prevailing percentage “61.969%” followed by palmitoleic acid (C16:1) “2.209%”. The content on polyunsaturated fatty acids “PUFA” in olive oil was “17.10%”, linoleic acid (C18:2) recorded “16.33%”, followed by linolenic acid (C18:3) “0.77%”, respectively. Olive oil contains the highest percent of MUFA, followed by SFA and PUFA, respectively.

In this respect, it was reported that [29], the percentages of oleic acid; palmitoleic (monounsaturated fats); linoleic; linolenic (polyunsaturated fats); palmitic; stearic and arachidic (saturated fats) in the olive oil were (71%, 1%, 10%, 1%, 13%, 3% and 1%), respectively.

The effect of two dosage of olive oil (0.25 ml and 0.5 ml/rats) and the two levels of Helianthus tuberosus L. (2% and 4%) and their effects together on consumption of diet/g for each rat/day, the % of body weight gain (BWG%), and the percentages of (liver & kidney) weights/body weight in diabetic rats recorded in Table 3.

The mean value of feed intake of diabetic group showed non-significant changes, as compared to the negative control group. Treating diabetic groups with the two levels of olive oil showed significant decrease in the mean value of feed intake p≤0.05, while feeding diabetic groups on diet containing (2% and 4% Helianthus tuberosus) induced non-significant changes, as compared to the positive control group. On the other hand, feeding diabetic rats on diet containing (2% and 4% Helianthus tuberosus) and treated daily with the two levels of olive oil (0.25 and 0.5 ml) led to significant increase p≤0.05, as compared to diabetic group (positive control group).

Injected rats with alloxan induced significant decrease in BWG%, as compared to healthy rats (control negative group). Treated diabetic groups daily with (0.25 ml and 0.5 ml olive oil) caused significant increase p≤0.05 in BWG%, as compared to the positive control group. Feeding diabetic group on diet containing 4% Helianthus tuberosus induced significant decrease p≤ 0.05 in BWG%, while the level of 2% Helianthus tuberosus showed non-significant changes in BWG%, as compared to the positive control group. Treating diabetic groups with (low and high levels from Helianthus tuberosus and the two dosage of olive oil) together increased the mean values of BWG%, comparing with positive control group.
Values in each column, which have different litters, are significant different (P < 0.05). The percentages of liver or kidney weights increased in the positive control group significantly p≤0.05, as compared to the negative control group. The weight % of these organs in all treated diabetic groups showed significant decrease p≤0.05, as compared to non-treated diabetic group (positive control group). Group of rats which treated with (4% Helianthus tuberosus and 0.5 ml olive oil/rat) recorded the best results in (liver and kidney weights/body weight%), followed by the group which treated with (2% Helianthus tuberosus and 0.25 ml olive oil/rat), respectively.
In this respect, it was reported that [30], Jerusalem artichoke “JA” (Helianthus tuberosus) has traditionally been known as a robust functional food. “JA” reduces the concentrations of glucose, triglyceride and the accumulation of cholesterol, thereby revealing its anti-obesity properties.

Research [31] reported that, fructo oligosaccharides present in Jerusalem artichoke “JA” (Helianthus tuberosus). Supplementation diet with “A” decreased liver weights by 18%, when compared to obese control group, thus reducing hepatomegaly. On the other hand, research [9] reported that, the weights of liver, kidney and heart not the spleen increased significantly in the diabetic group, as compared to the groups which were treated with inulin administration (p˂0.05). 

Table 4 illustrated the effect of Helianthus tuberosus and olive oil (alone or together) on serum glucose of diabetic rats. Injected rats with (150 mg alloxan) increased the mean value of serum glucose, as compared to the negative control group 157.595 ± 2.439 vs. 80.609 ± 3.093 mg/dl, respectively. All tested group which were treated with olive oil (0.25 ml and 0.5 ml/ each rat), Helianthus tuberosus (2% and 4%) and their combination (2% Helianthus tuberosus and 0.25 ml olive oil/rat) and (4% Helianthus tuberosus and 0.5 ml olive oil/rat) showed significant decrease p≤0.05 in serum glucose, as compared to the positive control group. The mean value of serum glucose decreased gradually with increasing the dosage of olive oil, Helianthus tuberosus and their combination. The highest decrease in serum glucose recorded for the group fed on diet containing 4% Helianthus tuberosus and treated daily with 0.5 ml olive oil/each rat. This treatment decreased the mean value of serum glucose by about 25.352%, then that of the positive control group.

In this respect, studies [32,33] reported that, Alloxan-treated rats led to decreased secretion of insulin by β-cells in pancreas and increased serum glucose level, therefore, the authors confirmed that, alloxan destroys pancreatic insulin secreting β-cells and induced hyperglycemia. Also, research [34] reported that, Alloxan induces diabetes by damaging the insulin secreting cells of the pancreas which causing to hyperglycemia.

Other studies [11,35] indicated that, treated rats with inulin improved glucose tolerance and lipid profile in the liver of rats fed a high-fat diet. The positive effect of Helianthus tuberosus tubers on cholesterol, triglyceride, HDL-c, LDL-c, VLDL-c and glucose can be attributed to their inulin content. Inulin is a water-soluble dietary fiber. Inulin can affect serum glucose and lipid profile by delaying gastric emptying or modifying the secretion of intestinal hormones such as glucagon-like peptide.

Supplemented diet with Jerusalem artichoke decreased blood glucose levels significantly in non-insulin dependent diabetes [36]. On the other hand, research [37] reported that, soluble dietary fiber plays an important role in controlling the concentration of glucose in the serum and decrease risk factors associated with diabetes. In this respect, research [38] reported that, inulin improves glucose tolerance and restores insulin secretion partially for diabetic patients during 4 to 6 weeks.

The effect of two levels olive oil, Helianthus tuberosus and (Helianthus tuberosus and olive oil together) on lipid profile of rats suffering from diabetes presented in Table 5. Data in Table 5 indicated that, the injection of rats with alloxan induced significant increase (p≤0.05) in all lipid profile, except high density lipoprotein-c HDL-c which showed significant decrease (p≤0.05), as compared to the control negative group. Diabetic groups which treated with Helianthus tuberosus, olive oil and (Helianthus tuberosus and olive oil together) with two levels, decreased the mean values of total cholesterol, triglycerides, low and very low-density lipoprotein-c significantly (p≤0.05), while HDL-c increased significantly, as compared to the positive control group.

The results in Table 5 showed that, all lipid parameters, except HDL-c decreased gradually with increasing the levels of olive oil, Helianthus tuberosus and (olive oil and Helianthus tuberosus together). On the other hand, HDL-c increased gradually. The best results in lipid profile recorded for the group, which treated with (4% Helianthus tuberosus and 0.5 ml olive oil/rat) with each other.

In this respect, research [39] showed that, total cholesterol, triglycerides, LDL-c increased significantly in diabetic rats, while HDL-c decreased.

The hypolipidimic effect of Jerusalem artichoke may be due to increasing fecal lipid excretion and decreasing lipid absorption [40,41]. On the other hand, studies [42] reported that, the hypolipeamic effects of inulin can be attributed to the action of fermentation products (i.e. short-chain fatty acids) of inulin in the large intestine by bifidobacteria. Short-chain fatty acids, mainly propionate, act as an inhibitor of hepatic lipid synthesis, including fatty acids and cholesterol.

Studies [43] reported that, the consumption 23 g olive oil/day decreased the risk factor of coronary heart disease (CHD) they may be due to the monounsaturated fat presence in olive oil. On the other hand, research [44] reported also, eating olive oil decreased serum cholesterol and LDL-c as compared to butter. In addition to, research [45] also reported, consumption of adults over 50 years of age (extra-virgin olive oil, about 2 tablespoons/ day) decreased total cholesterol and LDL-c for 6 weeks.

Phenolic compounds in olive oil, such as, hydroxytyrosol and oleuropein, led to inhibition of low-density lipoprotein-cholesterol (LDL-c) and high density lipoprotein-cholesterol (HDL-c) oxidation in vitro and in vivo, repress superoxide-driven reactions, and broke down the chain-like prevalence of lipid peroxides [46].

Table 6 illustrate the effect of olive oil and Helianthus tuberosus on Aspartate Amino Transferase AST, Alanine Amino Transferase ALT and Alkaline phosphatase ALP of diabetic rats. Injected rats with alloxan increased the mean values of AST, ALT and ALP significantly p≤0.05, as compared to the negative control group.

All treated diabetic groups with Helianthus tuberosus, olive oil and (Helianthus tuberosus & olive oil) together decrease AST, ALT and ALP significantly p≤0.05, as compared to the positive control group. On the other hand, increasing the levels of Helianthus tuberosus, olive oil and (Helianthus tuberosus & olive oil) together, decreased the mean values of liver enzymes gradually. Results in Table 6 showed that, Helianthus tuberosus with the two levels improved the mean values of AST, ALT and ALP, compared with the two levels of olive oil. The best results in liver enzymes recorded in diabetic group fed on diet containing 4% Helianthus tuberosus and treated with 0.5 ml olive oil (together), followed by the groups which were treated with (2% Helianthus tuberosus and 0.25 ml olive oil/rat) and the group treated with (0.5 ml olive oil/each rat), respectively.

Studies [11,36] reported that, treating (hyperglycemic or hyperlipidemic rats which induced by high fat diet) with Helianthus tuberosus HT tubers might improve the symptoms of these diseases, suggesting their potential as a useful functional food for prevent the progress of Non-alcoholic fatty liver disease and non-alcoholic steatohepatitis.

Jeruselem artichoke tubers “JAT” treatment significantly improved AST and ALT enzymes, as compared to CCl4 control group and succeeded to attenuate histopathological manifestations of hepatic fibrosis and lowered fibrosis score [47]. Hepatoprotectvie effect of JAE may be due to its antioxidant activity [48]. Oral administration of JAE might have a potential benefit of ameliorating diabetic symptoms by improving liver damage caused by STZ [49].
Research [50] reported that, the level of ALT enzyme decreased with diet containing olive oil, on the other side the high amount of olive oil (up to 20%) reduced total cholesterol in rats. Research [51] examined the effects of virgin olive oil on serum AST and ALT enzymes through a normal fat diet for 50 patients suffering from NAFLD. The authors reported that, a significant decrease in these enzymes was observed in the intervention group, as compared to the controls.

The effect of treating diabetic rats with olive oil, Helianthus tuberosus and (olive oil and Helianthus tuberosus together) on serum uric acid, urea nitrogen and creatinine presented in Table 7. The data in Table 7 revealed that, the mean value of serum uric acid, urea nitrogen and creatinine increased significantly (p≤0.05) in the positive control group, as compared to the negative control group.

Treating rats, which were, suffer from diabetes with the two dosage of olive oil, two levels of Helianthus tuberosus and (olive oil and Helianthus tuberosus together) induced significant decrease p≤0.05 in serum uric acid, urea nitrogen and creatinine, as compared to the diabetic control group. Kidney functions improved gradually with increasing the levels of tested diets. Diabetic rats which fed on diet containing high amount of Helianthus tuberosus and treated daily with 0.5 ml olive oil/rat recorded the best results in kidney functions, followed by the group fed on diet containing low amount of Helianthus tuberosus (2%) and treated daily with 0.25 ml olive oil/rat, respectively.

In this respect, research [52] reported that, the metabolic disturbance in diabetic animals may be elevated renal functions. Research [53] indicated that, feeding hyperglycemic rats on diet containing Jerusalem artichoke tubers would reduce serum glucose levels and improve lipid profile including (triglycerides, total cholesterol and LDL-cholesterol). The authors also reported that this treatment improved also liver and kidney functions. On the other hand, other studies [54,55] reported that, feeding rats on a diet containing inulin and oligofructose for some weeks decreased uremia, in both normal and nepherectomized rats. This may be due to, dietary inulin increased the fecal nitrogen excretion and reduced renal excretion of nitrogen in rats [56]. On the other hand, olive oil could have a beneficial role against oxidative and renal stress in rat, which induced by mercuric chloride induced [57].

CONCLUSION

It could be concluded that, Jerusalem Artichoke “Helianthus tuberosus L.” and olive oil (alone or together) improved the nutritional status and decreased the side effects of diabetic rats.
 
ACKNOWLEDGEMENT

The study was supported by the University of Hail.
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