Journal of Kerbala for Agricultural Sciences Issue (3), Volume (10), (2023)

Sept. 10, 2023 Abstract A field experiment was conducted at the second research station of the College of Agriculture, Al-Muthanna University, using a mixed soil. The objective of the study was to investigate the impact of varying quantities of biochar and organic fertilizer extract on the availability of nitrogen (N), phosphorus (P), potassium (K), iron (Fe), and zinc (Zn) elements in the soil .The experimental design encompassed the incorporation of four distinct biochar levels (A0, 0%), (A1, 1.5%), (A2, 2%), and (A3, 2.5%), which were applied proportionally to the soil volume. Additionally, four levels of organic fertilizer extract (0, 20, 40, and 80) liters per hectare (B0, B1, B2, and B3) were implemented. A factorial experiment was employed inside a randomized complete block design (RCBD) framework, consisting of three replications.The means were subjected to a Least substantial Difference (LSD) test at a significance threshold of 0.05. The findings indicated substantial disparities between the application of biochar and organic fertilizer extract, as well as their interaction, in relation to the levels of accessible nitrogen in the soil.Moreover, notable disparities were observed regarding the incorporation of biochar and organic fertilizer extract in relation to the accessibility of phosphorus within the soil. Furthermore, the introduction of biochar and organic fertilizer extract, as well as the interplay between these two factors, exhibited a substantial impact on the availability of potassium within the soil. Furthermore, the use of charcoal and organic fertilizer extract had a notable impact on the accessibility of iron and zinc within the soil.


Introduction
The kidney is an important organ in the body that is shaped like a bean, and is located in the abdominal cavity on both sides of the spine under the diaphragm, and the left kidney is at a higher level than the right kidney, and the kidney receives approximately 25% of the cardiac output, as the percentage of blood entering the kidney is in the region The cortex is about 85%, the outer medulla is about 14%, and the inner medulla is about 1% [1].The nephron represents the functional unit of the kidney, and one kidney consists of about one million nephrons and involves approximately 40 different types of cells [2].The basic function of the kidney is achieved through four main processes: Glomerular filtration (GF), Reabsorption, Secretion, and Excretion [3,4].as it works to purify the blood from waste products.With the glomerular filtration process, this function can be preserved even after losing half of the number of renal nephrons, and this is the reason why a healthy person can donate his kidney to lose 50% of the renal nephrons and maintain the normal glomerular filtration rate [5] as The kidneys perform many important functions in the body, the most prominent of which are maintaining the acid and alkaline balance, electrolyte balance, osmosis, removing harmful waste and maintaining the regulation of water in the body, furthermore secreting the hormone erythropoietin and cholesterol, and the kidney is one of the organs most vulnerable to toxic substances resulting from Medicines since the large volume of blood coming to them from the heart [6] Leea indica bandicoot berries have wide medical importance all over the world, as they are used to treat many health disorders.They have been used as a treatment for many skin problems, fever, joint pain and dizziness, furthermore being used to treat digestive disorders such as diarrhea and stomach ulcers [7,8], and it possesses anti-cancer, anti-microbial, antidiabetic, liver, vascular and central nervous system disorders, as well as used as a treatment for rheumatism, fractures, wounds, dysentery and sexual disorders [9], as well as the use of this plant to treat viral infections and some types of cancers Being one of the plants rich in antioxidants [10] as the leaves of this plant contain hydrocarbons, palmitic acid, phthalate derivatives, gallic acid, solanesol, phytosterols, and triterpenes, And catechins, tannins, flavonoids, coumarins, megastigmanes, and oxylipins.[11,12], and other studies demonstrated that the leaves of these plants contain other chemical components such as alkaloids, lignins, saponins, sterols, and anthraquinones [13] Diethylnitrosamine (DEN) is a yellow liquid organic chemical compound.It is considered a mutagenic and carcinogenic substance.It is classified as a nitrosamine with a chemical formula C4H10N2O.It has a boiling point of 177 degrees Celsius (351 F), a density of 0.95 grams / ml, and a molar mass of 102.14 grams / mol [14] It is a harmful pollutant that is widely found in many foodstuffs including salted fish, alcoholic beverages, milk, processed meat products, and many vegetables [15].In addition, it is present in tobacco smoke, cosmetics, agricultural chemicals, pharmaceutical drugs, and contaminated water.[16] Subject to food and environmental pollutants poses a significant risk to human and animal health.The harmful effects of DEN have been demonstrated to be increased production of free radicals, inflammation, and DNA damage.[17] It has been proven that DEN has carcinogenic effects on mice, hamsters, rats, guinea pigs, and rabbits, furthermore its carcinogenic effects on humans.Thus, DEN is widely used as an experimental carcinogen [18] Various nitrosamines are among the substances that cause acute hepatotoxicity and carcinogens in many types of animals, involving DEN.Which is given orally or by injection in experiments conducted on most rodents in high and effective doses for the purpose of inducing kidney tumors [19] DEN is primarily metabolized in the liver to its harmful metabolite, N-acetyl-p benzoquinoneimine, by cytochrome P450 [20] and particularly Cytochrom 2E1 enzymes, resulting in alkyl setcontaining metabolites that have The ability to cause DNA damage, promote programmed cell death, and eventually lead to injury to the kidneys and many other organs, involving the liver [21,17].The aim of the current study was to find out the protective role of Panicot berry leaves against nephrotoxicity resulting from diethylnitrosamine.

Materials and Methods
A total of 40 rats were used, divided into 4 groups (10 animals / set): the first set took a dose of 1 ml of plain water and is considered a control set, and the 2 nd set was injected with DEN at a concentration of 200 mg / kg, while the 3 rd set was given a daily dose of 60 mg / kg of Aqueous extract of bandicoot berries leaves, the IV set injected 200mg/kg of DEN + 60mg/kg of aqueous bandicoot berry extract.

Anatomy and take tissue samples
Animals were placed in a tightly closed transparent box after being given a chloroform-coated cotton swab to use for anesthesia.All rats from all sets were anesthetized and animals were dissected and these organs were cut into small pieces and preserved longitudinally and transversely and placed in 10% formalin for at least 48 hours.These samples were then dehydrated with ascending alcohol concentrations.After being cleaned with xylene, they were combined with paraffin wax at 56 °C to form blocks.All tissue sections were then stained with hematoxylin-eosin after placing the blocks and sections in a rotary microtome at 5 µm [22].

Preparation of the aqueous extract
A quantity of 20g of dry powder was used and subsequently combined with 400 milliliters of distilled water.The mixture was subjected to an electric grinder and allowed to stand at ambient temp for a duration of 24 hours.The mixture was subjected to filtration utilizing multiple layers of medical gauze, followed by centrifugation at a velocity of 3000 revolutions per minute.The solution was subjected to filtration utilizing filter paper for a duration of 10 minutes to achieve a clarified state.The extract was subjected to drying utilizing an electric oven at a temp of 40 degrees Celsius, and subsequently stored in a refrigerated environment until further use, as reported by [23].
The aforementioned technique was utilized to produce the Leea indica extract, albeit with certain alterations, outlined as follows: The desiccated foliage was pulverized utilizing an electric pulverizer and immersed in distilled water for a duration of 24 hours with continuous agitation.The resultant mixture was then filtered through multiple layers of medical gauze and soft, uncontaminated cloth.The filtrate was subsequently transferred to sanitized glassware and subjected to air drying via fan.The extract was then refrigerated until its intended use.

Results and discussion
The histological study demonstrated that the histological sections of the kidney of laboratory rats in the control groups, Figure (1).

Effect of diethyl nitrosamine treatment on kidney tissue
The findings of histopathology demonstrated that injection into the peritoneum with DEN led to congestion, atrophy, and shrinkage of the urinary glomerulus, furthermore an increase in Bowman's space, destruction of the urinary tubules, and sloughing of the inner lining of the urinary tubules.This is consistent with [24,25]The harmful renal effects that have occurred may be the result of subject to various exotic substances or medicines and toxins and the incitement of oxidative stress, and this may follow the activation of various factors and paths that participate in kidney tissue damage and increase the production of inflammatory cytokines, and these changes can lead to a imbalance between these The inflammatory factors and antioxidant defense systems of the cell, which ultimately leads to defrauding tissue disturbances and changes [26]The body subject to the substance of the DEN led to the effect of the kidneys and its tissues significantly as a result of the toxicity that occurs, and it occurs that the kidney injury is also related to the liver, given that the kidneys also participate in the DEN metabolism in the body, where the metabolism of the DEN occurs in the liver through the cytochrome system P450 It has become clear that generating the types of interactive oxygen ROS excessively causes kidney tissue damage by strengthening a peroxided fat and damaging cellular membranes, DNA and cellular proteins, as ROS causes an exhaustion of the antioxidant system, and the college is one of the most vulnerable organs to damage by many of the malware This may be since the availability of longterm multiple sebaceous fatty acids in the formation of renal fats and perhaps the tendon damage in the kidney is attributed to the activity of the stimulating infections such as the tnf-A tin-o tnf-A tin-o Secretion [27,28]It was also found that DEN decreases the activities of brush border membrane enzymes, increases lipid oxidation, protein oxidation, and increases hydrogen peroxide levels, thus destroying the cellular membranes of urinary tubules and glomeruli [29].

Effect of treatment with aqueous extract of leea indica leaves on kidney tissue
The present study's findings demonstrate that the leaves of Leea indica and its active compounds exhibit a beneficial effect on renal cell damage, renal filtration rate, and the functions and structure of glomeruli and renal tubules.Oxidative stress is known to result in a reduction of antioxidants, while the application of bandicoot berry botanical extract has been found to mitigate oxidative damage and safeguard kidney function enhancing the antioxidant system.According to [10,30] the enhancement of kidney tissue can be attributed to the active flavonoids found in the aqueous extract of the leaves.These flavonoids possess antioxidant properties, enabling them to donate hydrogen atoms to free radicals and mitigate their detrimental effects on the tissue.Consequently, this process decreases lipid peroxidation, safeguards cellular membranes against oxidative harm, and augments the tissue's repair system.Cellular damage [31]The reason for the improvement in the kidney tissue can be attributed to the fact that the bandicoot berry plant contains ascorbic acid, which works to protect cells from damage, as it is a catalyst for many enzymatic reactions, involving 2-oxoglutarate-dependent dioxygenase, which is important for the work and regulation of the embryonic structure of cells, through Stimulation of hydroxylation of nucleic acids DNA, RNA, as well as ascorbic acid maintains the stability of cells by protecting them from free radicals, by donating electrons to unstable molecules and converting them into stable molecules, decreases inflammation and repairs damaged tissues [32,33] The reason for the improvement in the kidney tissue may be since the fact that the aqueous extract of the plant contains megastigmanes, which has an important role in improving kidney function represented by decreasing levels of urea and creatinine in the blood, as well as increasing albumin levels and improving bilirubin, which leads to preventing the accumulation of more proteins in the tissues of the kidneys to decrease From renal damage, this findings in decreasing necrosis, congestion, and hemorrhage in the renal tissue, as well as decreasing glomeruli hyperplasia [34] It can be attributed to the preventive role of the raspberry leaves extract because it contains many active substances such as phenols and polyphenols that have the ability to sweep free radicals.The activity of the components of the antioxidant extract mainly since the properties of their oxidation and reduction.Breaking the series of free radicals by donating the hydrogen molecule and leads to the oxidation of the free root and its transformation into a stable condition, as a result of its external cover with electrons or the maintenance of the balance of antioxidants and the reduction of cancerous tumors resulting from the interactive oxygen types by visiting the super -desecotes activity (SOD) and glutathione (GSH) Also, by decreasing fat peroxide, gallic acid also stops the cell cycle, selfaccumulation, and programmed cell death by stimulating the cassepise path and working to regulate the proteins associated with the cell cycle such as Cyclin A, Cyclin D1 and Cyclin E, and delay the division of the unjust cell.By inciting the P27KIP enzyme and discharged CDK activity [35,36].

Figure ( 3 ):Figure( 4 ):
Figure (3):The effect of oral dose of 200 mg / kg of DEN for three months in male white rats was demonstrated, on the kidney tissue, as atrophy of the urinary glomerulus (black arrow) and shrinkage of the glomerulus (blue arrow) was observed, furthermore the breakdown of the urinary tubules ( Orange arrow) with hemorrhage (green arrow) furthermore an increase in Bowman's space (red arrow) , by hematoxylin and eosin 200 H&E stain