NEPHROPROTECTIVE OF ETHANOL EXTRACT OF Annona muricata L. BARK ON PARACETAMOL-INDUCED NEPHROTOXICITY IN RATS

Kidneys are organs that have the potential for damage caused by toxic substances. The use of paracetamol in high doses or long-term therapeutic doses can cause adverse effects in the form of damage to the kidneys. Plants that have the potential to become medicinal raw materials against kidney damage are Annona muricata L. The purpose of this study was to determine the potential protection of ethanolic extract of soursop bark (EESB) against histopathological rat kidney induced by toxic doses of paracetamol. This study used an experimental method using a completely randomized design (CRD). The normal control group (K0) and negative control (K) were given distilled water, while in the treatment group the doses of EESB were 150, 300 and 600 mg/kgBW dissolved in 1 ml of distilled water for 14 days. On the 7th day, all groups of rats were given paracetamol at a dose of 1350 mg/kgBW except for the K0 group. Each group was 5 repetitions, so that in this study used 25 rats ( Rattus norvegicus L.). Samples of kidney organs were taken on the 14th day to observe the histopathological structure of the tubular cells undergoing necrosis. The results showed that at a dose of 300 mg/kgBW EESB had the potential for protection against kidney histopathology by suppressing the occurrence of cell death (necrosis).


Tools and Materials
The tools that will be used in this

Research Process
The initial process of making

RESULTS AND DISCUSSION
This study used the bark of soursop (Annona muricata L.) which was processed using the maceration method.The type of solvent used in this study was 96% ethanol.Ethanol is used as a solvent because it is non-toxic, neutral, absorbs well, and can mix with water in all ratios and ethanol is a universal solvent that can attract polar, semi-polar, and non-polar compounds   where the antioxidant content is too high and also does not always have a good effect on the body.So that the optimal dose id needed to get a good effect as well.

CONCLUSION
Based on the results of this study, it can be concluded that from the three doses of ethanol extract of soursop bark (A.muricata L.) given, it was obtained that the administration of EESB at a dose of 300 mg/kgBW can provide potential protection against kidney damage by suppressing the level of damage that occurs in cells so that no cell death (necrosis).

Faculty
Paracetamol (acetaminophen) is one of the antipyretic analgesic drugs used by the community as a fever and pain-reducing drug [1].Paracetamol works by inhibiting prostaglandins that are weak in tissues (Asmara and Nugroho, 2017) [21].In several countriesor the use of therapeutic doses in the long term can cause toxic effects in the form of damage to the kidneys [3][4][5].The reactive metabolite formed from the metabolism of paracetamol is N-acetylp-benzoquinone imine (NAPQI) which can become a free radical that is both hepatoxic and nephrotoxic [7].The kidney is an organ that has the potential for damage caused by toxic substances.Blood circulation that enters the kidneys is 25-30% and will be cleaned by the kidneys.High blood flow, an increase in products excreted by the kidneys, and reabsorption of water in the tubules are the main factors affecting the sensitivity of the kidneys to toxic substances that enter the body [8].The main target of toxic substances in the kidney is the proximal tubule of the kidney [9].Structural changes in the kidney can be observed by looking at the cell injury that occurs, namely cell necrosis [8].The potential of a plant as a medicinal raw material in healing damage caused by a toxic substance can be seen from the antioxidant activity of the plant.One of the plants that have potential as medicinal raw materials is Annona muricata L. [10].mentions that the phytochemical content in the ethanol extract of soursop bark is alkaloids, saponins, tannins, flavonoids, and phenolics.The antioxidant effects of a plant can prevent an imbalance in the production of free radicals that can trigger oxidative stress, to prevent damage to the kidneys [112022.Preparation of ethanol extract of soursop bark (Annona muricata L.) (EESB) was carried out at the Organic Chemistry Laboratory, bark samples is collected and cut into smaller pieces.The bark is then dried.The process was carried out at room temperature without direct sunlight for 3 days.The preparation of ethanol extract of soursop bark was carried out using the maceration method.Soursop bark was soaked in a container containing 1 L of 96% ethanol solution for 3x24 hours.The next process was carried out using Wathmann filter paper to obtain the filtrate.The filtrate was concentrated using a rotary evaporator to obtain a thick extract.The extract was made in the form of a suspension in the form of a liquid extract dissolved with distilled water to facilitate administration to experimental animals.This study used 25 males Wistar rats aged 3 months with a body weight of about 200-250 g and acclimatized for 7 days.Rats are reared by giving them pellets and drinks ad libitum (to taste).The treatment in each group was administered orally for 14 days using a gavage needle.The determination of the dose of paracetamol and the determination of the dose of EESB in this study followed the dose given in the study [12].Groups K0 and K-were given distilled water, while group P1 was given EESB 150 mg/kgBW, P2 was given EESB 300 mg/kgBW and P3 was given EESB 600 mg/kgBW.Each feeding was given as much as 1 ml.On the 7th day, all groups of rats were given paracetamol at a dose of 1350 mg/kgBW in 1 ml of distilled water in one feeding except the normal control group.Kidney organ sampling in rats was carried out on the 14th day.Observation of kidney structure is done by performing histotechnical steps.Staining of renal histological samples was performed using hematoxylin and eosin (HE) staining.Kidney histology was observed with the aid of a light microscope at a magnification of 10 x 40.Kidney histology was observed with 3 times the microscope field of view in each incision.Observations were made by looking at the cortex of the kidney on tubular cells undergoing necrosis.Data on the structure of kidney cells undergoing necrosis in this study were analyzed using One Way ANOVA (Analysis of variance).If there is a difference in the effect of treatment, then the analysis is continued with the multiple distance test (Duncan) at the 5% level.

Figure 1 .
Figure 1.Diagram of the mean histopathological results of the kidneys