Research Article - Volume 1 Issue 1
Histomorphological effects of daucurs carota extract on dispirin induced kidney damage in albino rats
Ikem Amarachi Promise1; Okolie Nnamemeka1; Nnodim Johnkennedy1*
1Department of Medical Laboratory Science, Faculty of Health Science, Imo State University Owerri, Nigeria.
Received Date : Oct 21, 2021
Accepted Date : Nov 25, 2021
Published Date: Dec 02, 2021
Copyright:© Nnodim Johnkennedy 2021.
*Corresponding Author : r: Nnodim Johnkennedy, Department of Medical
Laboratory Science, Faculty of Health Science, Imo State University Owerri, Nigeria.
Email: johnkennedy23@yahoo.com
DOI: Doi.org/10.55920/2771-019X/1031
Abstract
This study was carried out to investigate the protective effect of extract against dispirin induced renal toxicity in albino rats.The histological effect of consumption of Daucurs carota root extract on the kidney of albino rats was carried out. The albinorats were divided into five groups. Each group consists of 5 albino rats. Group A serves as control while group B, C, D and E received dispirin 80 mg/kg/day, 50mg, 100mg and 200mg/kg body weight per day of Daucurs carota extract respectively for a period of 28 days. Serum samples from each rat were subjected to biochemical analysis for kidney functions, while kidney biopsies were also investigated by histological examination. Dispirin significantly increased serum creatinine, urea, (p < 0.05). Moreover, dispirin induced significant histological and ultrastructural alterations in the renal tissues of the rats. The exposure to Daucurs carota extract did attenuate the dispirin-induced toxicity in renal tissues. These results suggest that Daucurs carota extracts have significant protective role against dispirin-induced renal toxicity.
Keywords: Histomorphological; effects; daucurs carota; extract; dispirin; kidney damage; albino rats.
Introduction
The use of plants and plant extracts as trado-medical relief has been in existence for ages. This practice has been encouraged in recent times due to the less frequent side effects when compared with orthodox medicine. There has been growing interest in the investigation of the alternative route for the substitution of synthetic chemicals side effects, which are always in question [1]. Irrespective of the successes achieved using technology, the intuitive individuals across different geographical terrain especially developing countries still continue to use herbs for the treatment of certain ailments, either as a result of poverty or the efficacy of the herbal products. The use of herbal medicines and phytonutrients or nutraceuticals continues to expand rapidly across the world with many people now resorting to these products for treatment of various health challenges in different national healthcare settings. This past decade has obviously witnessed a tremendous surge in acceptance and public interest in natural therapies both in developing and developed countries, with these herbal remedies being available not only in drug stores, but now also in food stores and supermarkets [2].
As the global use of herbal medicinal products continues to grow and many more new products are introduced into the market, public health issues, and concerns surrounding their safety are also increasingly recognized. Although some herbal medicines have promising potential and are widely used, many of them remain untested and their use also not monitored. This makes knowledge of their potential adverse effects very limited and identification of the safest and most effective therapies as well as the promotion of their rational use more difficult. Daucurs carota is popularly known as carrot. Carrots are eaten raw in salads; cooked in soups, stir-fries, and casseroles; served alone as a side vegetable; and processed to make baby foods. Carrot is frequently used as a colourful garnish, often grated, sometimes sculpted into decorative flower-like shapes. Carrot juice is popular as a healthy drink, both alone or mixed with other vegetable and fruit juices. Carrots are used in sweet dishes such as carrot cake and in Asia often used in jams and syrups. Carrots are a source of the natural food dye carotene. Carrot fruit oil has been used for flavouring liqueurs and as an ingredient in cosmetics [3].
Essentially, herbal remedies consist of portions of plants or unpurified plant extracts containing several constituents which are often generally believed to work together synergistically [9]. Daucurs Carota are grown in the Northern Nigeria and used widely in Southern Nigerian states for various purposes such as treatment of infections and several ailments but it’s histopathological and effects on body tissues have been overlooked because of the properties and uses of these plants [4, 5]. Hence, this study is centered on the histopathological effects of Daucurs Carota on dispirin induced kidney damages in Wistar rats.
Materials and Methods
Animals
The twenty eight albino rats were bought from the animal house of College of Medicine and Health Science, Imo State University, Owerri Imo State. Albino rats of both sexes weighed between 180g to 200g maintaining under standard laboratory were used for the experiment. They were fed with grower mash bought from Farm Enterprises Nigeria Distributors of animal feeds, no 68 Mbaise Road Owerri .The present study was experimental, with the use of an animal model. The animal study was carried out in compliance with the policies outlined in the ‘Guide for the Care and Use of Laboratory Animals.
Plant Collection and Authentication
The root of Daucus carota were obtained at Ekeonunwa Market Owerri. It was identified by the Head Department of Plant Biology and Biotechnology, Faculty of Science, Imo State University Owerri, Nigeria. Extract Preparation; The roots of Daucus carota were kept at room temperature (29oC) in the Laboratory and pulverized into uniform fine powdered particle and sieved.. The dried down extract was reconstituted in distilled water for animal oral treatment by using 2g of the concentrated plant filtrate, dissolved in 20ml of distilled water to give a concentration of 100mg/ml
Study Design
Five groups of five male albino rats were used in the experiment. Each group was treated and fed as follows for four weeks:
Group 1: served as the normal control and received deionized water; Group 2: served as toxic control and received Dispirin solution; Group 3: Dispirin solution plus 50 mg/kg body weight Daucus carota; Group 4: Dispirin solution plus 100 mg/kg body weight. Daucus carota; Group 5: Dispirin solution plus 200 mg/kg body weight Daucus carota.
The dose of the Daucus carota extract used was administered by oral compulsion. The body weights were measured before commencement of the study and 28 days after administration.
On the 28th day, the rats were fasted overnight, weighed, and sacrificed under ether anesthesia on the 29th day. The blood samples were collected by cardiac puncture and kept at temperature of 4°C for 6 hours. The blood samples were then centrifuged at 3000 rpm for 15 minutes and stored properly for further analysis. The left kidney was stored in 10% formaldehyde and processed for histological examination.
Materials and Methods
Determination of serum creatinine by Jaffe’s Method and serumurea by enzyme methods [6].
Histology
For light microscopy examination, the formalin fixed tissues (kidney) were dehydrated through ascending grades of alcohol, cleared in three changes of xylene, and were embedded in paraffin. Serial sections, each of 4-micron thickness, were cut and stained with H and E as per standard protocol. Stained sections were morphologically evaluated, and the pictures of the slides were taken for comparison.
Tissue processing
At the termination of the experimental study, all the animals (treated and untreated) were sacrificed by (chloroform) anaesthetization. The visceral organs of interest (kidney) were excised and observed grossly. They were cut at 3-5mm and were processed by the use of an automatic tissue processor (Hestion-ATP7000 tissue processor-Germany) for dehydration, clearing, and impregnation. After embedding, sections were obtained at 3-5microns using digital rotary microtome (HestionERM 4000 Germany).
Photomicrography Mayer’s haematoxylin and eosin stained sections were examined using the Swift(R) binocular microscope® (Olympus England) with an in-built lighting system at x10 and 40 magnifications. Histopathological scoring was reviewed by two or more pathologist at the Federal Medical Centre Owerri. The study lasted between January, 2020 and March, 2020.
Photomicrography and Statistical analysis
Olympus photomicroscope (Opticshot-2; Nikon, Tokyo, Japan) was used at x40 magnification. Data were presented as Means ± S.E.M and significance were determined at p<0.05 using one way analysis of variance (ANOVA) and differences between pairs of means within the four groups of rats (treated and untreated) were made by using the Student’s test.
Results and Analysis
Weight of Albino rats treated with oral administration of different doses Daucus carota root extract and control for 28days.
Table 1: Initial weight (g) and final weight (g) of albino rats before and after 28 days.
Table 2: The mean level of serum creatinine and urea in albino rats administered with different doses of Daucus carota extract, control and dispirin.
Figure 1: Photomicrograph of the kidney of the control (CN). H&E ×400. Intact renal corpuscles with normal appearing glomeruli (G) and tubules (T), including the PCT (P) and DCT (D), are seen in control, as well as intact Bowman space (black arrow) and epithelial lining of Bowman capsule (arrowhead).
Figure 2: Photomicrograph of the kidney of the toxic control (Dispirin only) (TX). H&E ×400. Decreased cellularity in the glomeruli, loss of cellular constituents of tubules (double arrow), densely eosinophilic (‘colloid’) cast in the lumen of some tubules (dashed arrow) resulting in atrophy, loss of epithelia cells and severe cloudy swelling/inflammation of the distal convoluted tubules (yellow arrow) are observed in the toxic control group.
Figure 3: Photomicrograph of the kidney treatment with 50 mg/kg/day of Daucus carota extract. H&E ×400. Glomeruli (G) and surrounding Bowman's space (black arrow) are mostly intact. There is slight loss of cellular constituents of tubules and no eosinophilic (‘colloid’) casts are observed in the lumen of tubules.
Figure 4: Photomicrograph of the kidney treatment with 100 mg/kg/day of Daucus carota extract. H&E ×400. Glomeruli (G) and surrounding Bowman's space (black arrow) are mostly intact. There is slight loss of cellular constituents of tubules and no eosinophilic (‘colloid’) casts are observed in the lumen of tubules.
Figure 5: Photomicrograph of the kidney treatment with 200 mg/kg/day of Daucus carota extract. H&E ×400 (right). Glomeruli (G) and surrounding Bowman's space (black arrow) are mostly intact. There is slight loss of cellular constituents of tubules and no eosinophilic (‘colloid’) casts are observed in the lumen of tubules
Discussion
Daucus carota is used medicinally worldwide. The root and stem of the plant are used medicinally to treat different ailments. All parts of the plant have different ethno-medicinal uses for both man and animal alike. Medicinal plants are believed to be an important source of new chemical substances with potential therapeutic effects [7]. The kidney’s functional integrity is to maintain total body homeostasis through its role in the excretion of metabolic wastes and in regulation of intracellular fluid volume, electrolyte composition, and acid-base balance .This therefore implies that any harmful effect on body metabolism could be suggestive of toxic insult to the kidney [8].
In this study, administration of Daucus carota extract in rats caused significant increase in body weight as was noted in the experimental groups treated with Daucus carota extract. This may be as a result of the fact that Daucus carota extract induced inhibition of the intestinal motility, together with increase in the number of villi of the intestine and larger goblets cells, which favor rapid absorption of nutrient from the gastro-intestinal tract. This is in agreement with the report of [9]. Who reported that Daucus carota extract administered at different doses caused a significant increase in the body weight. It could also be due to its hypoglycemic property attributed to flavonoids and other phytochemical constituents of Daucus carota extract which promote food intake in albino rats The extract also contains major mineral elements, e.g. calcium, chloride, manganese, magnesium, zinc and potassium, which might also play a contributory role in enhancing its hypoglycemic property [6].
The results from this study indicate that aqueous extract of Daucus carota popularly has an increasing effects on the body weight of the experimental animals administered with dispirin. This significant increase in the body weights of the experimental animals was observed in a pattern suggesting the influence of the administered dosage [10].
The plasma creatinine and urea concentrations of the group that was administered with dispirin only were significantly higher than that of the control rats. These observed changes are in accordance with the report of [5] who reported that administration of dispirin caused marked impairment in renal function. The elevated concentration of plasma urea observed may have resulted from the inability of the glomeruli to filter urea. A significant decrease toward normal plasma level of creatinine and urea that was observed in the groups that received different doses of Daucus carota extract is an indication of tubular regeneration resulting from the release of reparative and prosurvival factors from the distal tubular cells. The marked decrease in plasma creatinine and urea in Daucus carota extract treated groups is in conformity with report of other researchers. This is an indication of improved blood flow to the kidney as well as repair of kidney tissue damage caused by dispirin. Therefore, this further suggests that Daucus carota root extract is not toxic to the histology of the kidney [11].
The histological observations of kidney sections from the groups that received dispirin only indicated some pathological states such as congestion of blood vessels, focal interstitial hemorrhage, and glomerular hemorrhages. This indeed is indication of renal tissue distortion suggestive of tissue damage in the group received dispirin only, whereas the histological appearance of the control group is consistent with normal histology. It can also be inferred that the activity of Daucus carota extract on the kidney is dose dependent considering the increase in the restoration of distorted kidney tissue with increase in the extract administration [12].
The Authors declare no conflict of interest
Conclusion
This study demonstrated the nephron-restorative effect of Daucus carota extract in dispirin induced nephrotoxicity in rats. The ameliorative effect of Daucus carota extract is evident by a remarkable restoration of markers of renal function such as urea, and creatinine levels in disprin treated rats.
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