# Introduction lcoholism and other serious health issues are brought on by excessive alcohol usage, including alcoholic liver damage (ALD). Alcoholism has been linked to several illnesses, and it is currently one of the most challenging health issues with substantial medical, social, and economic repercussions. (Pari and Karthikesan, 2001;. Alcohol abuse leads to significant illnesses such as hyperglycemia, cirrhosis, cardiovascular disease, pancreatic inflammation, and alcoholic fatty liver. (Ponnappa et al., 2000). Oxidative stress is one of the elements that are crucial in numerous pathways of alcohol-induced harm. The creation of ROS in our bodies is abnormally increased by our unhealthy eating habits and our way of life (smoking, drinking, obesity, and strenuous activity). When organisms experience oxidative stress brought on by free radical damage, antioxidants aid in coping. Antioxidant defenses come from two different sources: the diet, which includes fruits and vegetables, which are rich in vitamins C and E, carotenoids, ubiquinone, polyphenols, and lipoic acid. The other is endogenous and is made up of proteins, enzymes, or tiny molecules suchas glutathione, uric acid, superoxide dismutase, and glutathione peroxidase (ferritin, transferrin, etc.). Additionally, some elements that are significant cofactors include selenium, copper, and zinc. (Pincemail et al., 2009). We were particularly interested in the plant Moringa oleifera and the edible fungus Pleurotus ostreatus because they contain significant antioxidant content. The plant Moringa oleifera Lam. (Moringaceae), also known as Nebeday in Senegal, is of Indian ancestry and is now common throughout Asia and Africa. The leaves are utilized in traditional African medicine and are commonly consumed as a legume. They are an excellent source of protein (19-35% dry matter) (Kane et al., 2017;Makkar et al., (1996); Abou-Elezz et al., (2012) and are rich in metabolizable energy (2273-2978kcal/kg DM) (Makkar et al., (1996); Olugbemi et al., 2010). They A 11.2% dry Matter). The South African ecotype of the plant has been observed to contain 19.3% crude protein (Moyo., 2011). Traditional Chinese medicine uses M. oleifera leaves to treat diabetes, headaches, fever, and malnourishment (Ndong et al., 2007;Kerrarho., 1994). Preview studies have shown the health and nutritional interest of edible mushrooms (Zhang et al., 2016;Alam et al., 2008;Pornariya and Kanok, 2009). P. ostreatus has been demonstrated for its antitumor effects, antioxidant properties, antihyperlipidemic effects and antidiabetic effects, (Zhang et al., 2016;Abrams et al., 2011;Alam et al., 2008;Elmastas et al., 2007;Jayakumar et al., 2007;Jayakumar et al., 2006). A daily intake of 15 g of dried oyster mushrooms would have an anti-hyperlipidemic effect on the subjects, it would also cover up to 50% of the recommended daily intakes of macronutrients and minerals, according to research on Pleurotus ostreatus nutritional value and antihyperlipidemic effects on HIV-positive individuals taking ARVs (Abrams et al., 2011, Alam et al., 2008;Manzi et al., 2001;Alam et al., 2009;Manzi et al., 1999, Kane et al., 2017). Given the rich nutrient, phytochemical, and organoleptic potential of M.oleifera and P.ostreatus, we designed this study to determine the antihyperlipidemic effect of Moringa oleifera leaves and Pleurotus ostreatus in Wistar rats stressed by a combination of Ethanolparacetamol. In this paper, wewill code the dietary supplement by FMP16. # II. # Materials and Methods # a) Plant Material and Preparation of the mixture of Leaves from M. oleifera and P. ostreatus The fresh leaves of M. oleifera were harvested at the botanical garden of the University Cheikh Anta Diop (UCAD) of Dakar, Senegal and identified at the botanical department (UCAD).The leaves were cleaned immediately after harvest, cut into small pieces, and dried in the shade for two weeks. The dried material was ground into a powder using a manual homogenizer. P. ostreatus were obtained by cultivation at the biotechnological laboratory of the University Cheikh Anta Diop of Dakar. The Moringa oleifera and Pleurotus ostreatus powders were combined in a 2:1 ratio to create the dietary supplement. The mixture was created following Kane et al, instructions (2017). The combination was dissolved in 0.01% starch paste before being fed to the rats. # b) Animals and grouping Wistar rats strain to weigh 150 to 200 g were obtained from the Animal House of the National Institute of Youth and Sports in Yaounde. They were placed in plastic cages under standard laboratory conditions (temperature 20 to 30°C, relative air humidity 45 to 55%, and 12/12h light/dark cycle). The rats were fed with a basal diet and water ad libitum. The feed was a standard rat chow composed of carbohydrates (52%), protein (22%), fat (6.5%), water (12%), ash (6%), and fiber (4.5%). Every two days for 21 days, between 10:00 and 11:00 am, before the mixture administration of Moringa oleifera and Pleurotus ostreatus in proportion 2:1, made as reported by Kane et al., the amount of food and water ingested by each group of rats as well as body weights were recorded (2017). The experiments were performed during the day (09am-03pm). # c) Experimental design Thirty (30) adult male and female Wistar rats weighing 150 to 200 g were separated into five groups of six after two weeks of acclimatization: -Group 1 (TG): a stress-free control group that consumed only their regular diet of water, food, and vehicle (starch paste) once daily for 21 days, -Group 2 (TP): a control group that received paracetamol 12 hours after ethanol administration, was supplied in five sequential doses of 2 g. kg-1 using an orogastric tube to stress the group. For 21 days, they consumed the standard diet of water and food at their leisure in addition to the vehicle starch paste, -Group 3 (D1P): a group that received 500 mg/kg of FMP16 and was stressed by ethanol in five sequential doses of 2 g. kg -1 , administered through an orogastric tube; then received paracetamol 12 h after the last dose of ethanol. They received the standard diet (water and food ab-libitum) and the vehicle starch paste once a day for 21 days, -Group 4 (D2P): a group that received 1000 mg/kg of FMP16 and was stressed by ethanol in five sequential doses of 2 g. kg -1 , administered through an orogastric tube; then received paracetamol 12 h after the last amount of ethanol. They received the normal diet (water and food ab-libitum) and the vehicle starch paste once a day for 21 days, -Group 5 (D3P): a group that received 1500 mg/kg of FMP16 and was stressed by ethanol infive sequential doses of 2 g. kg -1 , administered through an orogastric tube; then received paracetamol 12 h after the last dose of ethanol. They received the standard diet (water and food ab-libitum) and the vehicle starch paste once a day for 21 days.The rats were given full access to food and water, and were on 12-hour light cycle each day (dark 12h-12h light). They were force-fed FMP16 using a gastroesophageal catheter and weighed every day. They fasted for the entire day before the animal sacrifice. On the 23rd day, the rats were given a night of rest before being slaughtered (while sedated with ether) by having their jugular veins cut. Organs such the liver, kidneys, brain, and testicles were collected along with blood. The liver, which was exclusively used in this processing day. It was then rinsed with ice-cold saline (0.9% NaCl) to eliminate any remaining blood. # d) Determination of the biochemicals parameters in liver -Preparation of liver supernates Prior to biochemicals analysis, each liver sample was homogenized using a Poterproctor placed on ice and 10% homogenate was prepared using the KCL buffer solution (1.15%). The homogenates were centrifuged at 3000 rpm for 30 min at 4 ° C to collect the supernatant used for analysis. The supernatant of each sample was aliquoted in 1. 5 # e) Statistical Analysis IBM SPSS Statistics 20 software was used for statistical analysis and data processing. P-values less than 0.05 were regarded as significant in the statistical analysis, which was conducted using one-way analysis of variance (ANOVA) and Bonferroni's post-test for multiple comparisons. The results are presented as the mean and standard deviation (SD). # III. # Results and Discussion Results have shown that no significant difference was observed in final body weights (155-173g) (Table1). Body weight gain ranged between 0.8 and 19 g for the four treatment groups. A decrease of 10% in the weight of D3N group was observed. These results corroborate those of Alam et al., (2011Alam et al., ( , 2009) ) who found that a diet enriched with Pleurotus ostreatus decreases the body weight of animals. However, Bobek et al., (1998) have shown that it does not affectthe weight as well as Schneider et al., (2011) who worked on humans with a daily dose of 30 g of dried oyster mushrooms, found that this does not affect anthropometric data. Bénissan et al., 2012, showed that the daily intake of 30 g of Moringa oleifera leaves improves nutritional recovery in children suffering from malnutrition. Hanaa et al., (2014), showed that a dose of 600 mg/kg of Moringa oleifera lowers the body mass index in obese subjects. Furthermore, the mixture of these species at a high dose of 1500 mg / kg, would explain the weight loss. This result was in contrast with those of Osman et al. (2012), who reported up to 14% changes in body weight of rats given M.oleifera extract for 21 days, attributing these changes to the rich nutrient quality of the extract. Results also have shown no significant difference in the amount of protein in the liver (figure1). Regarding lipid peroxidation (figure 2), results show no significant difference in the concentration of peroxidized lipids between the groups except between the unstressed control group (TG) and the 1500 mg/kg dose group where the concentration was 34% higher. These results are not in agreement with those of Mladenovic et al, (2013); Patere et al, (2011); Johnsen et al, (2007). The effect of FMP16 on oxidative stress enzymes such as catalase and cellular glutathione was also studied. Our results showed an increase in catalase activity of 87%, 85%, 90%, 82% respectively for the TG, D1P, D2P and D3P groups compared to the TP group (intoxicated and untreated). Also, catalase activity of the 1000 mg/kg dose (D2P) was 35% and 43% higher respectively compared to the 500 and 1500 mg/kg doses (figure 3 and 4). These results corroborate those of Lamou et ). The markers that are used to determine toxicity are usually transaminases (ALAT and ASAT), whose high concentration in the extracellular medium is synonymous with an alteration of the cells. In this study, our results showed that ALAT and ASAT activities were decreased in the FMP16 groups (Table 2). Thus, ALAT activity decreased by 29% in the 1000 mg/kg dose group compared to the intoxicated control. ASAT activity was higher in the intoxicated group (TP) by 28% and 26% compared to the 1000 and 1500 mg/kg doses. Compared to the results of the studies by Adedapo et al, (2009) and Alam et al, (2011), who instead found that a dose of 1600 mg/kg of Moringa oleifera leaves rather increased ALAT and ASAT activity; and on the other hand that a diet supplemented with 5% Pleurotus rather decreased transaminase activities, we could think that this explains the fact that FMP16 rather tends to regulate their activities due to the antagonistic effect that these two species have. Furthermore, results showed that the administration of FMP16 did not cause any significant difference in albumin and testosterone levels (Table 2). Regarding creatinine, FMP16 administration decreased creatinine levels in the treated groups (D1P, D2P, D3P) compared to the untreated and stressed group (TP) (Table 2). These results corroborate those of Sirag, (2009), Adedapo et al,(2009), Kane et al, 2022 who showed the protective effect of Pleurotus ostreatus and Moringa oleifera on kidney damage. Our results on lipid metabolism in rats revealed a significant decrease in total cholesterol in the 500, 1000 and 1500 mg/kg dose groups (figure 5). There was a 28%, 39%, 30% and 38% difference in TP, D1P, D2P and D3P compared to TG. In addition, a difference of 15% and 14% of D1P and D3P compared to TP. However, there was no significant difference in HDL cholesterol levels (Table 2). The results of the Triglycerides levels (Table 2) show a difference of 47% and 41% of the 500 and 1000 mg/kg dose compared to the TG control group. There was also a 28% decrease in Triglycerides levels at the 500 mg/kg dose compared to the dose 1000 mg/kg. In most of the studies on the effects of Pleurotus ostreatus and Moringa oleifera, they found a decrease in the concentration of LDL cholesterol which is more related to cardiovascular diseases (Bobek and Galbavy, 1999; Bobek et al., 1998;Hossain et al., 2003). These results are also in agreement with those of Alam et al, (2009), Schneider et al, (2011), Chumark et al, (2008), Kane et al, (2022). Our results (figure 6) and those of previous studies suggest that FMP16 would be an excellent cholesterol-lowering agent that could be recommended for the prevention and treatment of cardiovascular diseases. IV. # Conclusion A dietary supplement of Moringa oleifera leaves and Pleurotus ostreatus in wistar rats shows that the powders of M. oleifera leaves and P. ostreatus mixture have an antihyperlipidemic effect as it significantly lowers total and LDL cholesterol levels in rats stressed by combination of ethanol and paracetamol. The dose 1000 mg/kg is most appropriate for chemically stressed animals. FMP16 would have no effect on albumin and testosterone levels. 1GROUPES Starting Body weight (g) Final Body weight (g) P-value*TG154 ± 3,34173,67 ± 9,16 a0, 02TP154,33±3,44177,33±4,84 d0,01D1P154±2,53185,67±2,86 e0,01D2P153,20±2,68168±4,140,6D3P153±3,03157±7,140, 13*ANOVA test; TG: control group; TP: stressed group ethanol+paracetamol; D1P: stressed and treated group 500 mg/kg: D2P:stressed and treated group 1000 mg/kg; D3P: stressed and treated group 1500 mg/kg; a, d, e: mean statistically different with D3Pà p <0,05 (test de Bonferoni) 2triglycerides activityGROUPS ALAT (U/I)ASAT (U/I)ALBUMINE (g/dl)TESTOS. (ng/dl)HDL-C (mg/dl)Triglyc. (mg/dl)TG32,74±7,09160,92±30,021,53±0,200,31±0,0639,07±2,1852,03±0,68TP48,48±3,32196,35±33,401,61±0,370,47±0,1238,88±1,8878,14±12,69D1P43,16±8,80209,16±25,741,80±0,100,53±0,0836,30±2,8971,02±18,92D2P34,31±4,55141,86±11,191,60±0,200,44±0,0435,08±2,5398,18±7,64D3P43,66±2,13145,86±22,201,75±0,180,43±0,0334,17±5,4787,83±21,97 Year 202217Volume XXII Issue II Version ID D D D ) L(Medical ResearchGlobal Journal of© 2022 Global JournalsThe values are expressed as mean ± SD. TG: Control group rats with food and water ad libitum, TP: stressed rats without treatment, D1P: dose of 500 mg/kg, D2P: dose of 1000 mg/kg, D3P: dose of 1500 mg/kg. a, b ,c, d statistically different mean compared to TP, D1P, D2P and D3P at p<0.05 (LSD test) e, f mean statistically different from D1P and D3P at p<0.05(LSD test) The values are expressed as mean ± SD. TG: Control group rats with food and water ad libitum, TP: stressed rats without treatment, D1P: dose of 500 mg/kg, D2P: dose of 1000 mg/kg, D3P : dose of 1500 mg/kg. ## Acknowledgements The authors would like to thank the Biotechnology Laboratory of the University Cheikh Anta Diop of Dakar, Senegal, and the Faculty of Science, the FODRUS-LAPHER-Biotech of the University of Yaoundé I, Cameroon. The project was part of a scholarship provided to Fatou Corka KANE through Africa for Innovations, Mobility, Exchange, Globalization and Quality (AFIMEGQ) program sponsored by the European Commission's EACEA program. ## Conflict of Interests The authors declare that there is no conflict of interests regarding the publication of this paper. * Combating Nutrition with Moringa oleifera AMbora GMundia SMuasya World Agroforestry Centre 2004 * The nutritional effect of Moringa oleifera fresh leaves as feed supplement on Rhode Island Red hen egg production and quality Abou-Elezz FouadMohammed LSarmiento-Franco RSantos-Ricalde JFSanchez Tropical Animal Health and Production 44 5 2012 * Anti-hyperlipidemic effects of Pleurotus ostreatus (oyster mushrooms) in HIVinfected individuals taking antiretroviral therapy D IAbrams PCouey B SShade MEKelly N KElias PStamets BioMed Central Complementary and Alternative Medicine 11 2011 * Safety evaluations of the aqueous extract of the leaves of Moringa oleifera in rats AAdedapo O MMogbojuri Emikpe B O Journal of Medicinal Plants Research 3 8 2009 * Anti-ulcerogenic evaluation of the methanolic extracts of some indigenous medicinal plants of Pakistan in aspirin-ulcerated rats A HAkhtar KAhmad Journal of Ethnopharmacology 46 1 1995 * Nutritional Analysis of Cultivated Mushrooms in Bangladesh -Pleurotus ostreatus, Pleurotus sajor-caju, Pleurotus florida and Calocybe indica NAlam RAmin AKhan AraI JaShim M WoongLee M SooLee T Mycobiology 36 4 2008 * Ethanomedicinal knowledge of local Arab practitioners in the Middle East region HAzaizeh SFulder KKhalil OSaid Fitoterapia 74 2003 * Potential of mushroom bioactive molecules to develop healthcare biotech products MSBadalyan Proceedings of the 8th International Conference on mushroom biology and mushroom products (ICMBMP8) the 8th International Conference on mushroom biology and mushroom products (ICMBMP8) 2014 * Effect of Lactarius piperatus fruiting body maturity stage on antioxidant activity measured by several biochemical assays LBarros PBaptista IFerreira Cfr Food Chem.Toxicol 45 2007 * Nutritional characterization of Moringa (Moringa oleifera Lam.) leaves BMoyo PJMasika AHugo VMuchenje African Journal of Biotechnology 10 60 2011 * The, café: aliments ou médicaments DTallec La Phytothérapie Européenne 43 2008 * Temur N Determination of antioxidant activity and antioxidant compounds in wild edible mushrooms MElmastas OIsildak ITurkekul J. Food Comp. Anal 20 2007 * FAO (Food and Agriculture Organisation) The State of food insecurity in the world Internationals goals 2015 hunger reduction: uneven progress 2015 * The potential of Moringa oleifera for agricultural and industrial uses HN P SFoidl KMakkar Becker The Miracle Tree: Multiple Attributes of Moringa LJFuglie Dakar, Senegal 2001 CTA/CWS * Effects of intragastric administration of five oyster components on endurance exercise performance in mice GHao CZhang WCao JHao Pharmaceutical Biology 52 6 2014 * Knowledge and use of medicinal plants by local specialists in a region of Atlantic forest in the state of Pernambuco (North Eastern Brazil) LRGazzaneo PaivaDe Lucena RF PaulinoDe Albuquerque U J. Ethnobiol and Ethomed 2005 * Diversity of plant uses in two Caicaras communities from the Atlantic forest coast NHanazaki JYTamashiro HLeitao-Filho AGegossi Brazil. Biodiversity and conservation 9 2000 * Natural products and plants as liver protecting drugs SSHanda ASharma KKChakraborti Fitoterapia 57 5 1986 * Nutrional value and antinutritional components of whole and ethanol extracted Moringa oleifera leaves HP SMakkar KBecker Animal Feed Science and Technology 63 1-4 1996 * Antioxidant activity of the oyster mushroom, Pleurotus ostreatus, on CCl4-induced liver injury in rats TJayakumar RameshE GeraldineP Food Chem. Toxicol 44 1989-1996. 2006 * Protective effect of an extract of the oyster mushroom, Pleurotus ostreatus, on antioxidants of major organs of aged rats TJayakumar PAThomas GeraldineP Exp. Gerontol 42 2007 * JFuglie CTA 177 * CWS * Introduction to Moringa family MEOlson The Miracle Tree: The Multiple Attributes of Moringa LJFuglie Wageningen, The Netherlands; Dakar, Senegal 2001 CTA * Rapport de Synthèse, Direction de la Statistique et de la Prévision JKerrharo Enquête Démographique et de Santé II (EDSII) 1992/1993,1994 La pharmacopée Africaine, plantes médicinales et toxiques 1974. Ministère de l'Economie, des Finances et du Plan du Sénégal * KaneF C LTounkara DKimassoum MGuewo-Fokeng ATahir Diop WilfredF * Nutritional value of a dietary supplement of Moringa oleifera and Pleurotus ostreatus Mbacham African Journal of Food Science 11 6 2017 * Antioxidant Property of a Dietary Supplement of Moringa oleifera Leaves and Pleurotus ostreatus in Wistar Rats Subjected to Forced Swimming Endurance Test FCKane DKimassoum SFBrice MFPaul WFMbacham 10.4236/fns.2022.135037 Food and Nutrition Sciences 13 2022 * Huque R Effect of gamma radiation on antioxidant marker and microbial safety of fresh bitter gourd (Momordica charantia L.) AKhatun AHossain MIslam AHossain KMunshi Int. J. Biosci. (IJB) 2 11 2012 * Antifatigue Properties of the Aqueous Extract of Moringa oleifera in Rats Subjected to Forced Swimming Endurance Test BLamou G STaiwe AHamadou Abene JHoulray MAtou P VTan Antioxidant Oxidative Medicine and Cellular Longevity 2016. 2015 * Ethno-diversity within current ethnopharmocology as part of Israeli traditional medicine-A review LevE J. Ethodbiol and Ethnomed 2006 * Microsomal acetaldehyde oxidation is negligible in the presence of ethanol CSLieber YSWu KSSalmela Alcoholism 8 1994 * In vitro antioxidant activity and in vivo antifatigue effect of loach (Misgurnus anguillicaudatus) peptides prepared by papain digestion LJYou MMZhao JMRegenstein JYRen Food Chemistry 124 1 2011 * Introduction to the multiple uses of Moringa (7-10) LJFuglie The Miracle Tree : The Multiple Attributes of Moringa, L * Nutrition naturelle sous les tropiques LJFuglie L'arbre de la Vie : Les Multiples Usages du Moringa, L. J. Fuglie CTAetCWS, Dakar, Senegal 2002 * Bioactivity guided fractionation for anti-fatigue property of Acanthopanax senticosus L.-ZHuang B.-KHuang QYe L.-PQin Journal of Ethnopharmacology 133 1 2011 * Valeur nutritionnelle du Moringa oleifera, ´etude de la biodisponibilité du fer, effet del'enrichissement dedivers plats traditionnels senegalais avec la poudre des feuilles MNdong SWade NDossou ATGuiro RDGning African Journal of Food, Agriculture, Nutrition and Development 7 3 2007 * Evaluation of nutritional quality of moringa (Moringa oleifera Lam) leaves as an alternative protein source for Nile tilapia (Oreochromisniloticus L) NRichter PSiddhuraju KBecker Aquaculture 217 1-4 2003 * Okorie DO Chemical composition and nutritional value of mature and young fruiting-bodies of Pleurotus Pulmonarius produced on Andropogon gayanus straw and Khaya ivorensiss Sawdust ICOkwulehie JudithU IOSR J. Pharm. Biol. Sci 2014 * The effect of Moringa oleifera leaves on blood parameters and body weights of albino rats and rabbits HMOsman MEShayoub EMBabiker Jordan Journal of Biological Sciences 5 3 2012 * Protective role of caffeic acid against alcohol-induced biochemical changes in rats LPari KKarthikesan Fundam. Clin. Pharmacol 21 2007 * Ethnobotnical approaches of traditional medicine studies: Some experiences from Asia SJPei Pharmaceuticalbiology 2001 * Modeling alcohol's effect on organs in animal models BCPonnappa ERubin Alcohol. Res. Health 24 2000 * Amino acids and antioxidant properties of the oyster mushrooms, Pleurotus ostreatus and Pleurotus sajorcaju CPornariya OIKanok Sci. Asia 35 2009 * Antioxidant activity of proteins and peptides RJElias SSKellerby EADecker Critical Reviews in Food Science and Nutrition 48 5 2008 * Ethnobotany of Caicaras of the Atlantic forest Coast (Brazil) SCRossato HLeitao-Filho AGegossi Economic botany 53 1999 * Antioxidants and exercise SKPowers KHamilton Clinicsin Sports Medicine 18 3 1999 * Preventive effect of aqueous leaf extract of Alternanthera sessilis L. on CCl4 induced hepatic damage in albino mice ASivaraj PVinothkumar SPalani KDevi EKElumalai BSenthilkumar Int.J.Pharmagenesis 1 2 2010 * Hepatoprotective and antioxidant properties of Coccinia grandis aqueous leaf extract on ethanol induced liver toxicity in albino rats ASivaraj PVinothkumar KSathiyaraj KDevi BSenthilkumar J.Pharmacy.Res 3 3 2010 * Effect of Moringa (Moringa oleifera) inclusion in cassava based diets fed tobroilerchickens TSOlugbemi SKMutayoba FPLekule International Journal of Poultry Science 9 4 2010 * Hepatoprotective and antioxidant properties of aqueous rhizome extracts of Picrorhiza kurroa on CCl4 induced liver toxicity in albino rats PVinothkumar ASivaraj KDevi BSenthilkumar J.Pharmacy. Res 3 6 2010 * WHO (World Health Organisation) Report on the global situation of non-communicable diseases 2010 * Anti-diabetic effect of polysaccharides from Pleurotus ostreatus in streptozotocininduced diabetic rats YZhang HTao HZhou YZhang JinGYang Y Int. J. Biol. Macromole 83 2016