|Year : 2021 | Volume
| Issue : 2 | Page : 82-88
Hypolipidemic, antioxidant, and hepatoprotective effects of cucumber (Cucumis Sativus L.)-Supplemented diet in both sexes of sprague-dawley rats
Igbayilola Yusuff Dimeji1, Aina Olawale Samson2, Mofolorunso Adekunle Muiz1, Ashiru Mojeed Ayoola3, Morakinyo Ayodele Olufemi1
1 Department of Physiology, College of Medicine of the University of Lagos, Lagos, Nigeria
2 Department of Physiology, Lagos State University College of Medicine, Ikeja, Lagos State, Nigeria
3 Department of Biochemistry, College of Medicine of the University of Lagos, Lagos, Nigeria
|Date of Submission||14-Jan-2021|
|Date of Decision||27-Jan-2021|
|Date of Acceptance||27-Mar-2021|
|Date of Web Publication||10-Aug-2021|
Dr. Igbayilola Yusuff Dimeji
Department of Physiology, College of Medicine, University of Lagos
Source of Support: None, Conflict of Interest: None
Background: In Africa traditional medicine, certain plant leaves and fruits are employed in the treatment of metabolic disorders such as dyslipidemia, oxidative stress, and liver disorders. Cucumber (Cucumis sativus L.) is named among Nigerian plants that are under investigation for its medicinal activities. The present study investigates the hypolipidemic, antioxidant, and hepatoprotective potentials of C. sativus-supplemented diet in both sexes of Sprague-Dawley rats. Materials and Methods: Twelve male and twelve female Sprague-Dawley rats were used for this study and were grouped into four equal rats – Group A: control male (CM) fed with normal rat chow, Group B: control female (CF) fed with normal rat chow, Group C: cucumber-supplemented male (CSM) fed with cucumber-supplemented diet, and Group D: cucumber-supplemented female (CSF) fed with cucumber-supplemented diet for 6 weeks. At the end of the experiment, body weight change, cholesterol, high-density lipoprotein (HDL), triglyceride (TG), low-density lipoprotein (LDL), hepatic lipase, alkaline phosphatase (ALP), aspartate aminotransferase (AST), and alanine amino transferase (ALT) were determined. Hepatic and myocytic glycogen, superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), and malonaldehyde (MDA) were also assessed. Results: The result displayed a nonsignificant decrease (P > 0.05) in weight change in CSM and CSF compared with CM and CF. TG and LDL downregulated significantly (P > 0.05) in CSM and CSF compared with CM and CF while HDL significantly upregulated (P < 0.05) in CSM and CSF compared with CM and CF. There was a significant increase (P < 0.05) in SOD and CAT activities in CSM and CSF with a concomitant reduction in GSH activity and MDA level compared with CM and CF. ALP, AST, and ALT levels downregulated significantly (P < 0.05) in CSM and CSF compared with CM and CF. Despite a significant increase (P < 0.05) in skeletal glycogen, hepatic glycogen downregulated in CSM and CSF compared with CM and CF. Conclusion: It is evidenced that C. sativus-supplemented diet possessed hypolipidemic, antioxidant, and hepatoprotective effects in both sexes of Sprague-Dawley rats, though the effects were more marked in female rats compared with their male counterparts.
Keywords: Antioxidant, catalase, cucumber, medicinal, Sprague-Dawley
|How to cite this article:|
Dimeji IY, Samson AO, Muiz MA, Ayoola AM, Olufemi MA. Hypolipidemic, antioxidant, and hepatoprotective effects of cucumber (Cucumis Sativus L.)-Supplemented diet in both sexes of sprague-dawley rats. Niger J Exp Clin Biosci 2021;9:82-8
|How to cite this URL:|
Dimeji IY, Samson AO, Muiz MA, Ayoola AM, Olufemi MA. Hypolipidemic, antioxidant, and hepatoprotective effects of cucumber (Cucumis Sativus L.)-Supplemented diet in both sexes of sprague-dawley rats. Niger J Exp Clin Biosci [serial online] 2021 [cited 2022 Jun 26];9:82-8. Available from: https://www.njecbonline.org/text.asp?2021/9/2/82/323665
| Introduction|| |
The use of plants in traditional medicine referred to as herbalism or botanical medicine (Evans, 2002) falls outside the mainstream of Western or orthodox medicine. It has been estimated that about 75% of the world's population (mainly in developing countries) rely on traditional medicine as their primary form of health care. ]. The use of traditional medicine in the treatment and management of diseases in the Africa cannot fade away and this could be attributed to the socio-cultural, socio-economic of the traditional medicine due to lack of basic health care and qualified personnel.
Plants contain active components such as flavonoids, glycosides, saponins, and tannins, which possess medicinal properties that are harnessed for the treatment of different diseases. The active ingredients for a vast number of pharmaceutically derived medications contain components originating from phytochemicals. These active substances that contain the healing property are known as the active principles and are found to differ from plant to plant. The significance of plants in the treatment of diseases can be traced back to prehistoric times, and medicinal herbs are being increasingly studied by medical researchers (Sinclair, 1998). More than 80% of the world's population rely on herbal medicine for their primary health care, the majority of which use plants or their active principles. The use of plant resources mainly for traditional medicine, food, forages etc has been reported and in Nigeria, it represents a long history of human interaction with the environment and their in vitro and in vivo properties against microbial pathogens have been widely reported.,
Cucumber (Cucumis sativus) belongs to the family Cucurbitaceae. In general, there are 118 genera and 825 species worldwide among which 30 Cucumis species are found in Asia and African. Plants belonging to this family have a lot of medicinal and nutritional benefits. These may be as a result of the presence of some chemical substances that produce a definite physiological action on the human body. These chemicals are termed as phytochemicals. According to research, there is presence of the phytochemical called terpenoids in C. sativus extract. These particular phytochemical are known to possess medicinal potency against infections such as malaria, viral, bacterial and fungal infections. Evidences also corroborated the antioxidant effect of C. sativus in rats and substantial anti-inflammatory activity and anti-ulcer effect has also been reported. Saponins on the other hand have haemolytic property, induced cytotoxicity effect, antitumor and anti-mutagenic activities and can lower the risk of human cancers, by preventing cancer cells from growing. Phytosterols have also been found in the extracts of C. sativus (cucumber) and have been shown to have a significant hypocholesterolemic effect.
This study hypothesized that C. sativus-supplemented diet would possess hypolipidemic, antioxidant, and hepatoprotective effects in both sexes of Sprague-Dawley rats. In this study, we investigated both sexes of Sprague-Dawley rats exposed to C. sativus-supplemented diet to access the (1) impact on weight change; (2) determine the effect on lipid profile and hepatic lipase (HL); (3) access the effect on markers of oxidative balance such as superoxide dismutase (SOD), glutathione (GSH), catalase (CAT), and lipid peroxidation's (LPO's) MDA, and (4) whether the effect is sex dependent.
| Materials and Methods|| |
Twelve male and twelve female Sprague-Dawley rats were used for this study and were obtained from the animal house of the College of Medicine of the University of Lagos. The experimental procedures used were in accordance with the guidelines and provisions of the Animal Care and Use Research Ethics Committee (ACUREC) of the College of Medicine of the University of Lagos, Lagos State and the United States National Academy of Sciences Guide for the Care and Use of Laboratory Animals.
Fresh cucumber was obtained from a fruit seller in Kuto Market, Abeokuta, Ogun State. The cucumber obtained was peeled into smaller sizes and air-dried and grinded into powder form until constant weight of about 900 g was obtained. A small sample of the powdered form was taken to the laboratory for proximate, mineral, and vitamin analyses.
Approximately 850 g of dried cucumber was pelletized with 40 kg of normal rat chow to form the cucumber-supplemented diet (4.85 kg).
The experimental rats were allocated to one of four groups to be fed either a control diet or cucumber-supplemented diet. Food and water were available for all animals and grouped thus (six animals per group):
- Group A – Control male (CM) fed with normal rat chow
- Group B – Control female (CF) fed with normal rat chow
- Group C – Treated male fed with cucumber-supplemented male (CSM) diet
- Group D – Treated female fed with cucumber-supplemented female (CSF) diet.
Determination of body weight
The animals were weighed at birth and recorded at weekly intervals using a duet top loading weighing scale (Salter, England).
Collection of blood sample
Five milliliters of blood sample was taken by retro-orbital puncture. Blood was allowed to clot for 1 h at 4°C and then centrifuged at 3000 rpm for 10 min, and the serum samples were kept at −20°C until assayed.
Collection of tissue sample
After 6 weeks of the experiment, the rats were sacrificed using cervical dislocation. The animals were dissected; liver tissues were removed and washed in an ice cold, blotted, and weighed. A known weight of the organ was then homogenized with 0.1 M phosphate buffer (pH 7.2) with homogenizing machine. The resulting homogenate was centrifuged at 3000 rmp for 15 min. The samples were removed from the centrifuge, and the supernatants were decanted and stored at −20°C until analysis.
Cholesterol (CHOL), triglyceride (TG), high-density lipoprotein (HDL), and low-density lipoprotein (LDL) levels were carried out from the serum and liver homogenate samples with the aid of an automated analyzer (Mindray BS-120, Chema Diagnostica, Italy).
Determination of hepatic lipase (HL)
HL activity was determined in liver tissue homogenate thus:
The assay system (final volume 1 ml) contained 0.1 ml of glyceride emulsion, 0.2 ml of serum albumin, 0.6 ml of 0.1 M phosphate buffer (pH 7.4) and 0.1 ml of enzyme approximately 200 pg of lipase protein and was dissolved in glass-distilled water. The mixture was incubated for 60 min at 370C in a shaking water. Lipase activity was assayed by measuring the increase in absorbance at 546 nm bath.
Oxidative analyses of the serum and liver homogenate were assessed using the standard methods. Briefly, the most abundant individual aldehyde resulting from LPO breakdown in biological systems, malonaldehyde (MDA), was estimated. The reduced GSH content of the liver homogenate was determined while the activity of the SOD enzyme was determined. CAT activity was determined by measuring the exponential disappearance of H2O2 at 240 nm and expressed in units/mg of protein. Absorbance was recorded using Shimadzu recording spectrophotometer (UV 160) in all measurements.
Alkaline phosphatase (ALP), alkaline aminotransferase, and aspartate aminotransferase (AST) were determined by an automated analyzer (Mindray BS-120, Chema Diagnostica, Italy).
This was measured in the liver and gastrocnemius muscle samples (homogenate). Both tissues were harvested and cleaned immediately before the known weight was homogenized in ice-cold trichloroacetic acid (deproteinizing) solution and incubated for 15 min in a water bath. After discarding the precipitate, the supernatant was mixed with tetra sulfate IV acid and heated for 5 min. Absorbance was read with ELISA reader (Biobase Bioindustry Co. Ltd., Shandong, China) at 520 nm wavelength. A standard glycogen (Sigma; St. Louis MO, USA) was equally prepared and employed for the standard curve.
The results are presented as the mean and standard error of mean. GraphPad Prism Software (GraphPad, Inc., La Jolla, CA, USA) was used for statistical analyses. One-way analysis of variance with post hoc Tukey's multiple comparison test was used. The level of significance was set at P < 0.05.
| Results|| |
Result from the proximate analysis of dried C. sativus shows the presence of moisture (6.15), fat (3.36), ash (6.84), crude fibre (10.17), crude protein (10.96) and carbohydrate (62.58) [Table 1].
Vitamin compositions of dried Cucumber
Result from vitamin composition analysis of dried C. sativus shows the presence of water-soluble and fat soluble vitamins, A (32.016), B1 (0.058), B6 (0.021), C (2.673), D (0.111) and E (0.072). The results are presented in [Table 2].
Mineral compositions of dried Cucumber
Result from vitamin composition analysis of dried C. sativus shows the presence of mineral constituents which includes Ca (15.62), Mg (12.87), Mn (0.021), Fe (0.21), Na (1.20), Potassium (142.63), Si (0.01), Sulfur 90.060, Zn (0.18), Fluoride (1.29) and the results are presented in [Table 3].
Body weight changes
We examined body weight changes in male and female rats exposed to cucumber-supplemented diet. Although not significant (P > 0.05), [Figure 1] displays a progressive decrease in weight changes in CSM and CSF compared with their control counterparts.
|Figure 1: Effect of cucumber-supplemented diet on weight changes. Values represent mean ± SEM. CM: Control male, CF: Control female; CSM: Cucumber-supplemented male; CSF: Cucumber-supplemented female, SEM: Standard error of mean|
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[Figure 2]a shows a significant increase (P < 0.05) in SOD activity in CSF rats compared with their CM, CF, and CSM counterparts. GSH activity as displayed in [Figure 2]b downregulated significantly in CSM and CSF compared with CM and CF. Furthermore, a significant increase was observed (P < 0.05) in CSF compared with CSM. [Figure 2]c shows a significant increase (P < 0.05) in CAT activity in CSM and CSF rats compared with CM and CF. However, CAT activity upregulated significantly (P < 0.05) in CSF rats compared with their CSM counterparts. MDA level as shown in [Figure 2]d displayed a significant decrease (P < 0.05) only in CSF rats compared with CM and CSM rats.
|Figure 2: (a-d) Effect of cucumber-supplemented diet on GSH, SOD, and CAT activities and MDA level. Values represent mean ± SEM; Significant levels (*P < 0.05 vs. CM, αP < 0.05 vs. CF, βP < 0.05 vs. CSM). CM: Control male; CF: Control female; CSM: Cucumber-supplemented male; CSF: Cucumber-supplemented female, GSH: Glutathione, SOD: Superoxide dismutase, CAT: Catalase, MDA: Malonaldehyde, SEM: Standard error of mean|
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Hepatic and myocytic glycogen
[Figure 3]a shows a significant decrease (P < 0.05) in hepatic glycogen in CSF rats compared with CM, CF, and CSM rats while myocytic glycogen upregulated significantly (P < 0.05) in CSM and CSF rats compared with CM and CF rats. Furthermore, there was a significant elevation (P < 0.05) in CSF rats compared with their CSM counterparts [Figure 3]b.
|Figure 3: (a and b) Effect of cucumber-supplemented diet on hepatic and skeletal glycogen contents. Values represent mean ± SEM; significant levels (*P < 0.05 vs. CM, αP < 0.05 vs. CF, βP < 0.05 vs. CSM). CM: Control male, CF: Control female, CSM: Cucumber-supplemented male, CSF: Cucumber-supplemented female, SEM: Standard error of mean|
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There was a significant decrease (P < 0.05) in ALP level in CSM (8.55 ± 0.23) and CSF (7.53 ± 0.16) rats compared with CM (12.85 ± 0.39) and CF (6.53 ± 0.34) rats. ALP level upregulated significantly (P < 0.05) in CSM (8.55 ± 0.23) compared with CF (6.53 ± 0.34).
We also observed a significant reduction (P < 0.05) in AST level in CSM (197.5 ± 1.52) and CSF (153.8 ± 2.64) rats compared with CM (211.3 ± 1.26) and CF (205.5 ± 1.41) rats. However, AST level downregulated significantly (P < 0.05) in CSF rats (153.8 ± 2.64) compared with their CSM (197.5 ± 1.52) counterparts.
A significant decrease (P < 0.05) was observed in CSM (130.3 ± 0.76) and CSF (132.2 ± 0.42) rats compared with CM (135.3 ± 0.99) with a significant decrease (P < 0.05) in CSF (123.0 ± 1.13) rats compared with CSM (130.3 ± 0.76) rats [Table 4].
|Table 4: Effect of cucumber-supplemented diet on alkaline phosphatase, aspartate aminotransferase, and alanine aminotransferase levels|
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There was no significant difference (P > 0.05) in CHOL level in CSM (0.75 ± 0.02) and CSF (0.72 ± 0.02) rats compared with CM (0.77 ± 0.03) and CF (0.42 ± 0.03) rats. CHOL level upregulated significantly (P < 0.05) in CSM (0.75 ± 0.02) and CSF (0.72 ± 0.02) rats compared with CF (0.42 ± 0.03) rats.
We observed a significant reduction (P < 0.05) in TG level only in CSF (0.28 ± 0.03) rats compared with CM (0.60 ± 0.03), CF (0.59 ± 0.04), and CSM (0.55 ± 0.03) rats.
A significant increase (P < 0.05) was observed in HDL level in CSM (0.24 ± 0.01) and CSF (0.20 ± 0.02) rats compared with CM (0.06 ± 0.01) and CF (0.02 ± 0.01) rats with a significant decrease (P < 0.05) in CSF rats (0.20 ± 0.02) compared with CSM 0.20 ± 0.02 rats.
LDL level downregulated significantly (P < 0.05) in CSM (0.41 ± 0.03) and CSF (0.50 ± 0.02) rats compared with CM (0.66 ± 0.03) rats [Table 5].
|Table 5: Effect of cucumber-supplemented diet on cholesterol, triglyceride, high-density lipoprotein, and low-density lipoprotein levels|
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HL activity displayed a significant increase in CSM and CSF rats compared with their CM counterparts [Figure 4].
|Figure 4: Effect of cucumber-supplemented diet on hepatic and skeletal glycogen contents. Values represent mean ± SEM; Significant levels (*P < 0.05 vs. CM). CM: Control male; CF: Control female; CSM: Cucumber-supplemented male; CSF: Cucumber-supplemented female, SEM: Standard error of mean|
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| Discussion|| |
C. sativus (cucumber) has been described as a suitable functional food for medicinal purposes and has been employed for the treatment of metabolic disorders such as diabetes, hyperlipidemia, and hypertension (as diuretic) and for the treatment of gallbladder stones, constipation, and dyspepsia in Asian traditional medicine. The significance of serum lipids in atherosclerosis and coronary heart disease has been proved by many researchers. Emerging evidence has demonstrated the cardiovascular disease risk to be positively associated with TC and TG and inversely associated with HDL.,
Hyperlipidemia is a heterogeneous group of disorders characterized by high level of lipids in the bloodstream. It may be caused by disorders of some endocrine glands, kidneys, effects of certain drugs, dietary intake containing a high amount of fat, risky lifestyle, and aging. It is one of the risk factors in the development of atherosclerosis. In the current study, decreased LDL and increased HDL levels were observed suggestive of the hypolipidemic effect of C. sativus. It was also observed that the C. sativus possesses hypotriglyceridemic effect with a significant reduction of TG both in male and female rats. This result was in line with the study carried out by Hu et al. on the effects of bioactive components of cucumber on the lipid profile and lipid absorption, which revealed that the treatment significantly reduced the serum TG.
Fiber has been reported to decrease LDL by interrupting CHOL and bile acid absorption and increasing LDL receptor activity. LDL has been shown to facilitate the transport of CHOL into a cell; thus, the significant reduction in LDL is suggestive and justifies the CHOL-lowering effect of C. sativus observed in the current study, thereby further reducing the risk of metabolic disorder such as obesity and atherosclerosis. There have been reports on the lipid profile of various plants including C. sativus and some of which are in accordance with the present study., Thus, there could be alterations in the concentration of the various lipid metabolism and predisposition of the heart to atherosclerosis and its associated coronary heart diseases. Furthermore, the presence of phytosterol in Cucumis sativus fruit has been reported. Phytosterols have a significant hypocholesterolemic effect and CHOL-lowering potentials. Therefore, the CHOL-lowering effect of C. sativus fruit homogenate could also be attributed also the presence of phytosterols. Reduction of LDL concentration may be due to the antioxidant property of C. sativus.
There was a significant reduction in HL in male and female rats exposed to C. sativus-supplemented diet. Recent studies have demonstrated that the liver contains lipases (glycerol-ester hydrolase) capable of hydrolyzing glycerides. It has been reported that elevation of the lipoprotein lipase (LPL) activity results in reducing TG level in the blood and reduction of the plasma TG level and elevation of the plasma HDL CHOL by activation of LPL may be useful for the prevention of dyslipidemia. Hypertriglyceridemia and HDL-hypocholesterolemia are well-known risk factors for atherosclerosis. Balanced CHOL level reduces the incidence of LDL oxidation and the associated risk of atherosclerosis and other related heart diseases. Liver injury causes the accumulation of abnormal amounts of fats, predominantly triacylglycerol in the parenchymal cells into the systemic circulation. In addition, decreased liver enzymes observed in both male and female rats showed a possible hepatoprotective effect of C. sativus, and this is in agreement with the previous study.
Oxidative stress results when the antioxidant system is overwhelmed by the generation of excess reactive oxygen species. These reactive species such as superoxide radical anion (O− 2), hydrogen peroxide (H2O2), and hydroxyl radicals (HO−) cause severe damage to macromolecules, tissues, and organs through the process of LPO, protein modification, and DNA strand breaks. Oxidative stress resulting from the generation of these free radicals is known to contribute immensely to several pathological conditions such as aging, cancer, cardiovascular disorder, and neurodegenerative diseases among others. In this study, the SOD and GSH properties were investigated because of their synergistic ability to work hand in hand. SOD has been shown to catalyze the breakdown of superoxide, the most common free radical in the body into oxygen and hydrogen peroxide, while GSH catalyzes the breakdown of hydrogen peroxide to water.
Furthermore, C. sativus produced a significant increase in SOD and CAT activities suggestive of reduced production of oxidative radicals from the current study. This is in agreement with a previous study which reported that the presence of antioxidant and antimicrobial properties, its minerals (especially iron), Vitamins (especially Vitamin A and C), and high protein contents was found to prevent oxidative radicals' production. This could also be due to the presence of secondary metabolites such as saponins and terpenoids. A significant reduction was observed in MDA's LPO level in all the doses tested suggestive of effective oxidative balance. MDA has been shown to be a biomarker of oxidative stress, excessive production of which has been linked to dyslipidemia and atherosclerosis.
| Conclusion|| |
In conclusion, it is evidenced that cucumber-supplemented diet possessed hypolipidemic, antioxidant, and hepatoprotective effects in both sexes of Sprague-Dawley rats, though the effects were more marked in female rats compared with their male counterparts.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
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[Figure 1], [Figure 2], [Figure 3], [Figure 4]
[Table 1], [Table 2], [Table 3], [Table 4], [Table 5]