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Abstract Liver disease is a chronic disease in many countries of the world, due to lack of effective protective / therapeutic option for the disease. It has been reported that a high percentage of Egyptian society has liver disease, which is mainly due to chronic hepatitis C virus (HCV), in addition to other risk factors including obesity, diabetes, excessive medication and environmental pollution. Oxidative stress is a common mechanism involved in the initiation and progression of liver damage in different types of liver disease. It has been found that the use of natural antioxidants is useful in the plan of logical therapeutic in the prevention and therapeutic of liver diseases including oxidative stress. There is a trend in the search for natural antioxidants of natural sources that have a high efficiency in the protection and therapeutic of liver diseases. Moringa Oleifera Lam (M.O) a plant native to Indian, and then spread to many parts of the world, and the different parts of this plant have been used in ancient Indian civilization as an important food source and in the therapeutic of some diseases, and recently found that the different parts of the plant contain high levels of vitamins and minerals that are useful in food, in addition to containing antioxidants such as Phenols and flavonoids, which can be used in the protection or therapeutic of diseases that involve oxidative stress. Acetaminphen (Paracetamol) APAP is most widely used in the world as an analgesic and antipyretic drug that is safe at therapeutic dosages. Paracetamol is available as oral, rectal and injectable formulation. Paracetamol is prescribed as pain reliever and fever reducer. It is commonly used to relieve headache and other minor muscle aches, pain and major ingredient in numerous cold and flu remedies, and broadly use in the management of more severe pain such as post-surgical pain and providing palliative care in advanced cancer patient. The use of Paracetamol at recommended doses is safe. However, its use at high doses causes significant damage to the liver. Therefore, the models of liver damage induced by Paracetamol in rats and mice are widely used to study the effects of antioxidants in liver protection. This study aimed to: (1) Evaluation of antioxidant activity of different parts (leaves, flowers, stem bark, roots, seeds) of Moringa Olifeira plant extracts in-vitro and to identify the parts containing high antioxidant activity (2) Evaluation of ethanolic extracts of leaves and pods of Moringa Oleifera as protective against paracetamol-induced liver damage (acetaminophen -APAP) in rats. In this experiment, the ethanolic extract, for leaves or stem bark of the Moringa (300 mg in H2O + DMSO / kg of rats weight or 600 mg in H2O + DMSO / kg of rats weight) were given to the rats one hour before to administration of a single dose (oral) of acetaminophen (APAP) dissolved in DMSO (4g/kg bw of the rat). Protective activity of Moringa extracts of leaves and pods was followed weekly for 21 days to monitor liver functions enzymes such as alanine aminotransferase (ALT) and oxidative stress markers such as Malondialdhyde (MDA), Catalase (CAT) and the histological changes in the liver were compared between groups of rats treated with Moringa extract with a group of rats treated with acetaminophen alone. In this experiment, kidney function markers (urea and creatinine) were also measured. As oxidative stress in the liver is accompanied by oxidative stress in the kidney. For this purpose male albino 42 rats were divided into seven groups (each group has 6 rats) as the below:group I: Normal control (receiving water (W) daily). group II: Rats received (DMSO daily) control negative. group III: Rats received a singl dose of 4g APAP/kg b.w once (positive control). group IV: Rats received 4g APAP/kg b.w once one hour after receiving the M.O extract (1mL) 300 mg of M.O leaves extract/kg b.w, daily, for 21 days group V: Rats received 4g APAP/kg b.w once one hour after receiving the M.O extract (1mL) 600 mg of M.O leaves extract/kg b.w daily, for 21 days group VI: Rats received 4 g APAP/kg b.w once one hour after receiving the M.O extract (1mL) 300 mg of M.O stem bark extract/kg b.w daily, for 21 days group VII: Rats received 4 g APAP/kg b.w once one hour after receiving the M.O extract (1mL) 600 mg of M.O stem bark extract/kg b.w daily, for 21 days The most important results obtained include the following elements: 1. Evaluation of the antioxidant activity (In -vitro) using the DPPH for different parts of Moringa using three solvents: ethyl alcohol, water and hexane. The results shown that the ethanolic extracts gives the highest efficiency of the antioxidant activity of all parts of the plant. In addition, it was found that the extract of flowers gave the highest efficiency of antioxidant activity (94.91%) followed by leaves extract (93.10%), stem bark (90.78%), roots (84.80%) and finally seeds (62.15%). This is compared to Vit C (ascorbic acid) which used as reference compound given (94.00%) 2. High-efficiency liquid chromatography – Mass spectroscoby (HPLCMS) analysis for ethanolic extract of leaves detect five phenolic and flavonoid compounds, including chlorogenic acid, Apigenin-8-C— glucoside, Quercetin-3-O-β-D-glucoside, Quercetin-3-O-acetyl glucoside and kaempferol-3-O-glucoside. These compounds are known to have anti-oxidant activity. Although there were other compounds in the ethanolic extract of leaves, it was not possible to identify them because there were no reference compounds for comparison. 3. In the biological experiment in-vivo, the ethanolic extract of both leaves and pods in doses (300 mg / kg of rats weight and 600 mg / kg of rats weight) was given orally one hour before to the preoperative dose of paracetamol (APAP) (4g / kg) of the rats. The preventive activity of these extracts was studied, by monitoring the level of liver function markers, oxidative stress markers and histopathology in the liver, weekly for 21 days. The most important results obtained are: (a) In the group of rats treated with acetaminophen (APAP) alone (positive control or hepatotoxin control) there was a significant increase ( p ≤ 0.05 ) in liver function enzymes ALT, AST, GGT, ALP in serum at the first week and continued to increase until the end of the experiment (at the third week) compared to the normal control group. The change in these enzymes indicates damage to the liver tissue due to oxidative stress of paracetamol (APAP). (b) In the group of rats treated with ethanolic extract of leaves and stem bark in doses 300 and 600 mg / kg of rats weight (Groups 4 through 7) the results showed a significant decrease (p ≤ 0.05) in the levels of these enzymes during the three weeks compared to the group of positive control (paracetamol treatment alone). This indicating the protective effects of both ethanolic extract of leaves and stem bark, where the levels of these enzymes returned to normal level at the end of the experiment. In addition, this reduction depends on the amount of the dose of the extract.(c) The results also showed a significant increase in serum total bilirubin and a significant decrease in serum total protein during the three weeks in the group treated with paracetamol (APAP) alone. These results also indicated liver tissue damage due to paracetamol effects. On the other hand, the treatment with the ethanolic extracts of both leaves and stem bark returned these parameters to their normal level, which indicates their protective effects on the liver. 4 - The study of the protective effect of the ethanolic extracts of leaves and stem bark of Moringa on the oxidation markers that including lipid oxidation (LPO) using the test of Malondialdhyde (MDA) and the enzymes of the antioxidant superoxide dismutase (SOD) and catalase (CAT).The results showed the following: (a) In the paracetamol(APAP) group alone, the MDA level in the blood increased after a week of treatment and continued to rise steadly to the end of experiment. In comparison, the treatment of ethanolic extracts of leaves and stem bark of Moringa showed a significant decrease (p ≤ 0.05) in MDA during the experiment period and the decrease reached a level slightly lower than the normal control at the end of experiment. In addition, the treatment with solvent DMSO led to a small decrease compared to normal control. (b) Treatment with paracetamol(APAP) significantly reduced SOD and CAT at the end of the experiment, indicating oxidative stress. On the other hand, the protective effects of both the ethanolic extract of leaves and stem bark against the oxidative stress of paracetamol (APAP) showed a high level of these enzymes to a level higher than both the positive control and the normal control. 5. Histological changes. The histological examination of the liver for different treatments showed the following: The normal control group showed no histopathological changes in the liver, tissues at the end of the experiment. The control group for DMSO also showed no histopathological changes in the liver, tissues at the end of the experiment. The positive control group (APAP treatment alone) showed a focal death in the liver with inflammation of the cells, in addition to hyperplasia in the superficial surface of the vertebrae, at the end of the experiment. The group treated with ethanolic extract of leaves and stem bark showed slight improvement in these histological changes, at the end of the experiment. from these results we can concluded the following: 1- The ethanolic extracts of Moringa oleifera Lam leaves and stem bark showed antioxidant activities (both in- vitro and invivo) due to the presence of active phytochemical such as phenols and flavonoids and other active compounds . 2- The ethanolic extracts of Moringa Oleifera Lam leaves and stem bark exhibits free radical scavenging and hepatoprotective and kidney protective against oxidative stress injury. |