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ORIGINAL ARTICLE
Year : 2020  |  Volume : 10  |  Issue : 3  |  Page : 149-153

The Dosage of the Avocado Leaf Extract (Persea americana Mill.) on Regeneration of Diabetic White Rats (Rattus norvegicus) Renal Cell


1 Faculty of Public Health, Nutrition Department, Tadulako University, City of Palu, Indonesia
2 Chemistry Education, Faculty of Teacher Training and Education, Tadulako University, City of Palu, Indonesia

Date of Submission23-Jan-2020
Date of Decision20-Feb-2020
Date of Acceptance14-May-2020
Date of Web Publication20-Aug-2020

Correspondence Address:
Nurdin Rahman
Soekarno Street Km 9, City of Palu, Indonesia
Indonesia
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/ijnpnd.ijnpnd_6_20

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   Abstract 


Context: Antioxidant activity in avocado leaves plays a role in improving insulin work in controlling blood glucose levels. Aim: To analyse the effect of doses of avocado leaf extract on the regeneration of diabetic rat renal cells. Methods: Experimental method was used with 18 male white rats as subjects. White rats were divided into six groups: Treatment G1: Feed + STZ (Streptozotocin) + Sucrose 10% + Avocado leaf extract dose of 100 mg/kg body weight + NaCMC 0.5%, G2: Feed + STZ + Sucrose 10% + Avocado leaf extract 150 mg/kg body weight + NaCMC 0.5%, G3: Feed + STZ + Sucrose 10% + Avocado leaf extract dose 200 mg/kg body weight + Na-CMC 0.5%, G4: Feed + STZ + 10% sucrose + Na-CMC 0.5%, G5: Feed + STZ + 10% sucrose + Glibenclamid + Na-CMC 0.5%, and P6: Feed. Results: The average renal damage of rats in the rat group was G1 = 1.66; G2 = 1.33; G3 = 0; G4 = 3; P5 = 0; P6 = 0. Score 0 = no damage. Score 1 = mild renal cell damage (1/3 cell part); score 2 = moderate renal cell damage (2/3 cell parts); score 3 = severe renal cell damage (>2/3 cell parts). Conclusion: The treatment of G3 with a dose of 200 mg/kg body weight avocado leaf extract gives a relatively effective influence on regenerating STZ induced renal cell rats, compared to other doses.

Keywords: Avocado leaf extract, diabetes mellitus, renal hispatology


How to cite this article:
Rahman N, Sabang SM, Dewi NU, Bohari, Fitrasyah S. The Dosage of the Avocado Leaf Extract (Persea americana Mill.) on Regeneration of Diabetic White Rats (Rattus norvegicus) Renal Cell. Int J Nutr Pharmacol Neurol Dis 2020;10:149-53

How to cite this URL:
Rahman N, Sabang SM, Dewi NU, Bohari, Fitrasyah S. The Dosage of the Avocado Leaf Extract (Persea americana Mill.) on Regeneration of Diabetic White Rats (Rattus norvegicus) Renal Cell. Int J Nutr Pharmacol Neurol Dis [serial online] 2020 [cited 2020 Oct 26];10:149-53. Available from: https://www.ijnpnd.com/text.asp?2020/10/3/149/292688




   Introduction Top


Diabetes mellitus is a deadly disease and causes complications such as hypertension, stroke, coronary heart disease, kidney failure, cataracts, glaucoma, destruction of the retina of the eye causing people blind, impotence, impaired liver function, long period of healed wounds, and infection that must be amputated.[1] The process of diabetes mellitus that occurs from time to time can cause damage to the body’s systems, especially nerves and blood vessels, which are also the main causes of kidney failure.[2],[3] Kidney failure caused by diabetes is known as Diabetes Nephropathy, which is the biggest cause of kidney failure.[4]

The condition of chronic hyperglycemia will result in increased production of free radicals resulting in oxidative stress.[5] Oxidative stress is a condition in which free radicals in the form of reactive molecules, which occur through a biochemical reaction from normal cells, damage cell membranes, and cause various bodily dysfunctions.[6],[7] Free radicals in the body will cause damage to DNA, carbohydrates, proteins, and lipids[8] Under the normal circumstances, the formation of free radicals will be followed by the formation of antioxidants in the body resulting in a balance of free radicals and antioxidants.[9] The condition of a molecule where there are more free radicals than antioxidants will cause cell damage. The most appropriate step to reduce oxidative stress is by reducing free radicals or optimizing body defense by increasing antioxidant intake

Antioxidants are agents that are efficient in preventing lipid peroxidation and protect cells from oxidative stress-mediated by Reactive Oxygen Species.[10] Medicinal plants are considered as a source of various oxidative stress-related diseases.[11] Phenolic compounds are usually found in medicinal plants and food products and mainly consisted of phenolic acids, flavonoids, and tannins. These compounds have a wide range of antioxidant activities.[12] Most colored plants, fruits, and vegetables contain antioxidants. One of them is avocado leaves, which phenolic composition related to free radical capture activity.

This study is aimed at analyzing the effects of avocado leaf extracts on the regeneration of diabetic rat renal cells


   Material and Method Top


Experimental method was used with 18 male white rats as subjects. This research was carried out in the Pharmacology Laboratory of Pelita Mas Palu STIFA and Balai Besar Veteriner Maros.

Avocado leaf extract

Avocado leaves were extracted with 70% of positive ethanol solution containing flavonoid compounds, saponins, tannins, steroids. The 70% concentration is an avocado leaf extract that gives absorbance value and optimum percent inhibition

Groups of treatment

The rats are grouped into six groups, the different dosages administered, and the behavior monitored. White rats were divided into six groups as follow:
  1. G1: Feed + STZ (Streptozotocin) + Sucrose 10% + Avocado leaf extract dose of 100 mg/kg body weight + NaCMC 0.5%,
  2. G2: Feed + STZ + Sucrose 10% + Avocado leaf extract 150 mg/kg body weight + NaCMC 0.5%,
  3. G3: Feed + STZ + Sucrose 10% + Avocado leaf extract dose 200 mg/kg body weight + Na-CMC 0.5%,
  4. G4: Feed + STZ + 10% sucrose + Na-CMC 0.5%,
  5. G5: Feed + STZ + 10% sucrose + Glibenclamid + Na-CMC 0.5%, and
  6. P6: Feed.


Tested animal surgery

The surgical process is performed on the abdominothoracal part then necropsy of the kidney organ is carried out. The organ is then rinsed with 0.9% of physiological liquid of NaCl to separate it from the blood or fats attached to the organ.

Research ethics

This study has obtained an ethical recommendation agreement Number: 546/H4.8.4.31/PP36-KOMETIK/2018 issued by the Health Research Ethics Committee of the Faculty of Medicine RSPTN of Hasanuddin University, and RSUP Dr. Wahidin Sudirohusodo Makassar.

Data analysis

Primary data collected in this study includes the results of examination of the histopathological picture of renal male white rats in the form of histopathological scoring, namely score 0: normal, score 1: renal tubular damage for about 1–25%, score 2: renal tubular damage for about 25–50%, score 3: damage to renal tubules for more than 50–75%, and score 4: damage to renal tubules for more than 75–100%. Data are obtained in the form of kidney damage scoring for each treatment.


   Results Top


The results showed that the average renal damage of rats in the rat group was G1 =1.66; G2 =1.33; G3 = 0; G4 =3; P5 = 0; P6 =0. Score 0 = no damage. Score 1 = mild renal cell damage (1/3 cell part); score 2 = moderate renal cell damage (2/3 cell parts); score 3 = severe renal cell damage (>2/3 cell parts) [Table 1] and [Figure 1].
Table 1 Scoring results of observation of kidney histology

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Figure 1 The level of damage of kidney cells in various groups

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Avocado leaf extract at a dose of 100 mg/kg the body has no effect on kidney tissue regeneration in male white rats (1.67). The average score for kidney damage is reduced to 1.33 when the dose is increased to 150 mg/kg of the body. Furthermore, rats in group 3 who received a dose of 200 mg/kg body weight had an average score of kidney damage 0. The score was the same as group five who received glibenclamide.


   Discussions Top


Based on the results of the study on rats histopathological preparations, it was proven that multilevel dosing of avocado leaf extract had an effect on regenerating kidney tissue of male white rats, but at a dose of 100 mg/kg, it did not have an effect on regenerating kidney tissue, and at a dose of 150 mg/kg of body weight, it gave an effect on regenerating kidney tissue but not too good compared to the dose of 200 mg/kg of body weight. The greater the dose of avocado leaf extract, the greater the regeneration of renal tubular cells, while the smaller the dose of avocado leaf extract, the smaller the regeneration of renal tubular cells that are damaged due to the induction of ethylene glycol. It is shown that the greater the dose of avocado leaf extract, the more the amount of active compounds contained in the extract so that the regeneration of kidney tubule cells is getting bigger.

Other studies showed that moringa leaf extract can improve renal histopathology of experimental wistar rats with diabetic mellitus in the form of a decrease in the amount of fatty degeneration and necrosis, and a dose of 400 mg/kg body weight had the best effect on repairing kidney damage.[13] A study conducted by Shofia[14] states that diabetic mellitus treated with brown seaweed (Sargassum prismaticum) therapy containing flavonoids, which is an antioxidant that functions as a free radical scavenger, can suppress the formation of ROS which causes tissue damage providing tissue repair as indicated by narrow distance between bowman and glomerular capsules.

Other studies show that white rats induced by alloxan with doses of red betel leaf extract of 100 mg/kg of body weight containing active ingredients in the form of flavonoids and alkaloids that are antioxidants from red betel leaf with a dose of 100 mg/kg of body weight (P3) give a microscopic picture (degeneration and necrosis) of lighter kidney so that cells can be repaired or regenerate cells suffering from necrosis.[15]

Based on the observation of histopathological preparation in the kidney, there is damage in the form of cell degeneration and inflammation in renal tubular epithelial cells. Cell degeneration is a damage that occurs in the cytoplasm, but it does not damaging the cell nucleus so that the cell can recover to normal if the cause of the damage is gone. Cell degeneration that lasts long and continuously will result in inability of the cell in carrying out metabolism properly, and cell will be broken. This is an initial reaction to acute cell damage caused by ischemia or toxin. In a degenerative state, the cell can regenerate and return to function as before if the toxin compound causing the damage is removed. Cell degeneration is caused by the accumulation of toxic substances and other metabolites.[16]

Renal tubular repair and regeneration of epithelial cells are believed to be closely related to the content of phenolic compounds.[17] Medicinal plant antioxidants are able to reduce ameliorate oxidative induced kidney damage by lipid peroxidation and enhancement of a scavenging ability of antioxidant defense system.[18] Avocado leaves contain phenolic compounds such as alkaloids, flavonoids, saponins, tannins, and polyphenols, according to the results obtained in the phytochemical screening test which shows the presence of these compounds in avocado leaf extract.[19] Compound that is believed to have an effect on diuretic activity is flavonoid.[20] Flavonoid can increase urinalysis and electrolyte secretion in mice by increasing glomerular filtration rate (GFR).[21] High glomerular filtration speed (GFR) makes the kidney able to get rid of waste products and toxic substances from the body. Furthermore, the polyphenol compounds contained in avocado leaves help fight free radicals in the body as well as prevent the inflammatory process in the body cells. Flavonoid is a compound that is very effective in repairing and protecting cell structures, as well as an anti-inflammatory.[22],[23]The implication of the results of this study is that avocado leaf extract can be used and developed as a source of antioxidants that play a role in the regeneration of kidney cells that degenerate due to high blood glucose levels.


   Conclusion Top


The treatment of G3 with a dose of 200 mg/kg body weight avocado leaf extract gives a relatively effective influence on regenerating STZ induced renal cell rats, compared to other doses.

Acknowledgements

The authors are thankful to the Directorate General of Development and Research Enhancement of the Ministry of Research, Technology and Higher Education for research funds.

Financial support and sponsorship

The authors are thankful to Directorate General of Development and Research Enhancement of the Ministry of Research, Technology and Higher Education.

Conflicts of interest

There is no conflict of interest in the preparation and completion process of this research.



 
   References Top

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Kamaliani BR, Setiasih NLE, Winaya IBO. Histopathological kidney overview of experimental diabetes mellitus wistar rats given ethanol extract of moringa leaf. Buletin Veteriner Udayana 2019;71. doi:10.24843/bulvet.2019.v11.i01.p12  Back to cited text no. 13
    
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Shofia V, Aulanni’am A, Mahdi C. Studi Pemberian Ekstrak Rumput Laut Coklat (Sargassum Prismaticum) Terhadap Kadar Malondialdehid dan Gambaran Histologi Jaringan Ginjal Pada Tikus (Rattus Norvegicus) Diabetes Melitus Tipe 1. Jurnal Ilmu Kimia Universitas Brawijaya 2013;1:119-25.  Back to cited text no. 14
    
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