Pa ge 1 Pa ge 49 American Journal of Multidisciplinary Research and Innovation (AJMRI) Hepatoprotective Activity of Plant Extracts in Non-Human Models: The Toxicants Solvents and Diseased Models-A Review Dongsogo Julius1*, Larbie Christopher2, Idrissu Abdul Mumeen3, Abera Ataanya Daniel4, Daniel Tuurisoe5 Volume 2 Issue 6, Year 2023 ISSN: 2158-8155 (Online), 2832-4854 (Print) DOI: https://doi.org/10.54536/ajmri.v2i6.672 https://journals.e-palli.com/home/index.php/ajmri Article Information ABSTRACT Received: October 15, 2023 Accepted: November 18, 2023 Published: November 23, 2023 The search for an ideal hepatic regenerative agent has led to the modelling of several forms of liver disorders. This involves the usage of various substances in in-vitro and in-vivo systems that are injurious to the human liver in in-vitro and in-vivo systems and then challenged with plants parts extracted with organic and inorganic solvents. Reported view works on hepatotoxicity have been focused on the plant species, this review therefore focused on the toxicants, the solvents for extraction and models used in inducing the hepatotoxicity. Google search, Elsevier and PubMed databases were searched for primary articles from 2005-2022 on hepatotoxicity. In all, 206 articles were retrieved, of which 46 were rejected and 165 were included for the analysis. Authors reported 55 toxicants used in inducing 10 in-vitro and in-vitro models and challenged with 94 different medicinal plants species extracted using 11 different organic and inorganic solvents. Comparatively, tetrachloromethane (CCL4) was the most frequent toxicant 55 of 94 (58.5%) reported plant species while ethanol 25 (45.5%) was the most frequently used solvent for extracting plant phytoconstituents. For the models used in inducing hepatotoxicity, Wistar albino rats 46 of the 55 (83.6%) reported toxicants was the most predominant. The liver is prone to many available toxicants however, phytohepatoregeneration is possible with many medicinal plants which can easily be extracted with common solvents. Keywords Phytohepatoregeneration, Toxicants, Hepatotoxicity, Hepatic INTRODUCTION The human liver carries majority of metabolic activities necessary for maintaining homeostasis, cell growth, and repairing degraded, worn-out cells and tissues (Jamuna et al., 2018; Eghba et al., 2019; Elzwi, 2019). The liver is located in the lower quadrant below the diaphragm (Madrigal-Santillan et al., 2014; Al-Snafi et al., 2019). The liver is also responsible for synthesizing essential amino acids, fatty acids, vitamins, bile, immunoglobulins which are necessary for proteins synthesis, membrane and immune protection against diseases and infections (Elmansi et al., 2017; Maha, 2019; Koubaa et al., 2020). The liver also detoxifies harmful products of digestion and respiration such as xenobiotics, toxins and drugs into harmless metabolites for excretion by the kidney and skin (Onojia et al., 2019; Olajide et al., 2020). Bile produced by the liver also emulsifies fats from food, making it easier for lipase action during digestion (Partel et al., 2019). The hepatic portal vein connects the liver to the heart where it receives oxygenated blood from the heart and in turn supply blood with glucose, vitamins, proteins and nutrients necessary to the heart (Usunomena et al., 2015; Sani et al., 2020; Uchenna et al., 2021). The numerous and multifaceted functions of the liver make it a target for many toxins and pathogens which often results in hepatic diseases such as hepatitis, hepatoma, hepatomegaly, fibrosis, hepatocellular carcinoma, hepatosteatosis and hepatobiliary disorders (Akpanyung et al., 2019; Akharaiyu and Okafor, 2021). Therapeutic drugs such as paracetamol, tramadol, rampicin, Adriamycin, caffein as well as environmental pollutants such as carbon tetrachloride produced charged intermediate metabolites which in excess above the body antioxidant capacity results in oxidative stress (Ige et al., 2017; Ajiboyea et al., 2018; Aly et al., 2020). Sustained oxidative stress initiate chain hepatocyte peroxidation of membrane lipids, oxidize sulfhydryl group containing proteins of enzymes and alkylate hepatic cells DNA resulting loss of membrane permeability, inhibition of sensitive enzymes action and necrosis (Elzwi, 2019; Mohammed et al., 2020). Though the idiopathic phase of all liver diseases is cirrhosis or carcinoma, the pathophysiology depends on the toxicant’s nature or infectious agent’s nature (Zahra et al., 2012; Sadashiva et al., 2019). Pathogenic microorganisms such as hepatitis B and C viruses usually cause insertional mutagenesis of their genome into hepatic cell DNA (Okaiyeto et al., 2018; Okoro, 2020). This mutagenesis results in the translation and transcription of untargeted proteins resulting in undesirable physiological actions (Okaiyeto et al., 2018; Okoro, 2020). The infected hepatocytes also display the viral surface antigens resulting in ‘’unself ’’ recognition by the body immune surveillance systems (Senti et al., 2016; Okoro, 2020). Chronic hepatic 1 Department of Biochemistry and Molecular Biology, Faculty of Biosciences, University for Development Studies, Tamale, Ghana 2 Department of Biochemistry and Biotechnology, Faculty of Biosciences, Kwame Nkrumah University of Science and Technology, Kumasi, Ghana 3 Department of Biochemistry, Faculty of Biosciences, University for Development Studies, Tamale, Ghana 4 Department of Medical Laboratory Technology, Kumasi Technical University, Kumasi, Ghana 5 Laboratory Department, Tamale Teaching Hospital, Tamale, Ghana * Corresponding author’s e-mail: jdongsogo@gmail.com Pa ge 50 https://journals.e-palli.com/home/index.php/ajmri Am. J. Multidis. Res. Innov. 2(6) 49-56, 2023 infection initiates the expression of proinflammatory and pro-fibrogenic cytokines which cause necrosis, apoptosis and autophagy of infected hepatocytes (Hassan et al., 2015; Oyibo et al., 2020; Nwaogu et al., 2022). Sustained fibrosis result in replacement of degraded hepatocytes with hepatic stellate cells containing extracellular matrix proteins such as collagen and alpha muscle actions which finally lead to cirrhosis (Usmani et al., 2019; Acheampong et al., 2021). Heavy metals such mercury (Hg), Cadmium (Cd), Arsenic (Ar) and lead (Pb) also react with cellular lipids, carbohydrates and proteins to produce reactive oxygen, nitrogen, superoxide, peroxide and molecular oxygen species beyond the body glutathione and antioxidants enzymes (glutathione peroxidase, superoxide dismutase, catalase) capacity (Chinnula et al., 2018; Ogunmoloye et al., 2022). The loss of antioxidant/free radical balance results in oxidative stress (Dkhil et al., 2013; Iwo et al., 2017). In oxidative stress, the transcription factor nuclear factor kappa Beta NF-kB) is activated in the hepatocyte cytosol and then translocated into the nucleus of the hepatocytes where it causes the gene overexpression of inflammatory cytokines including transforming growth factor 1(TGF-1), tumor necrosis factor-alpha (TNF- alpha), interleukins 1 and 8 (Madrigal-Santillan et al., 2014; Lee et al.,2017; Hashem et al., 2019; Ezzat et al., 2020). The profibrogenic cytokines also induce the expression of apoptotic mediators such as caspases-9 and antimitotic cyclins which induced apoptosis of hepatocytes (Immih et al.,2022). Diagnostic and prognostic indicators of liver diseases in clinical and experimental settings rely on liver function markers such as transaminases (alanine transaminase and glutamine transaminase), proteins (globulins and albumins) and pigments (bilirubins) (Olatosin et al., 2014; Mirazi and Karami, 2016; Syvenia et al., 2018). Serum alkaline phosphatase levels indicate membrane disruption (Ashraf et al., 2018). Inflammatory and fibrogenic cytokines TNF-ᾳ, transforming growth factor (TGF), nuclear factor kappa B (NF-kB), inducible nitric oxide synthase, cyclooxygenase-2 (COX-2) and interleukins 1,6,8,10 are indicated by PCR quantification of their mRNA levels (Rehab et al., 2016; Ramirez-Marroquin et al., 2019; Salih et al., 2022) as markers of inflammation and fibrogenesis. Oxidative stress is indicated by reduced glutathione level (GSH) and the enzymes glutathione peroxidase, catalase and superoxide dismutase while peroxidation of lipids and proteins are measured using malonaldehyde (MDA), thiobarbituric reactive substances (TBARS), myeloperoxidase (MPO), p-carbonyl proteins, 4-HNE proteins and paraoxonase (Achuba et al., 2019). Mitotic inhibition is indicated by p-cyclins while bax and bcl-2 proteins indicate apoptosis levels (Ameaka et al., 2021). Hepatic steatosis is indicated by mRNA expression of ᵞ-PPAR and CAT-1 while epithelial-mesenchymal transition (EMT) indicate progression from fibrosis to carcinoma (Elmansi et al., 2017; Ghareeb et al., 2019). Plant phytochemicals offers promising alternative for the discovery of ideal hepatorestorative agent to replace pharmaceutical drugs which are unavailable, expensive and have harmful side effects (Ameaka et al., 2021). The quest to identify this liver drug have led to the modelling of liver diseases termed hepatotoxicity in animals such as mice, rats and tissues and then challenged with plant parts such as root, leaves, bark, wood, fruit, flowers, puds, sap, fibre and twigs are extracted with organic and inorganic solvents such as alcohols, petroleum ether, water (Sarfo- Antwi et al., 2018; Tokofai et al., 2020). Hepatotoxicity activity of several medicinal plants have been reported, objective of this review to produce a one- stop collection of these toxicants, their diseased models and solvents. MATERIALS AND METHODS Google scholar, Elsevier and PubMed databases are searched for journals from 2005-2022 on hepatotoxicity using the key words; hepatotoxicity, botanical name of plants and the toxicants. Inclusion into the study is the availability of the plant species, substance for inducing the toxicity, extraction solvent and animal/tissues used in the toxicity. Review articles and journals earlier than 2005 were excluded. Quality checks on the articles downloaded were performed using standard protocols. Statistical Analysis The information was extracted on Microsoft Excel version 2020 (Microsoft Incorporated, New York) and reported as graphs and tables. Discrete data are reported in percentages. RESULTS AND DISCUSSION Number of Journals Retrieved from Databases In all, 206 peer reviewed articles were downloaded of which fewer 41 (19.9%) were rejected while majority 165 (80.1%) were included in the review. Toxicants Challenged with the Plant Species In all, 55 toxicants have been reported against 94 plant species. Of these, CCL4 is the most reported toxicant used against 55 plant species, paracetamol/acetaminophen followed with 27 plant species being tested against it. Ethanol (50-100%) has been tested against 12 plants while thioacetamide and dimethylnitosamine are both tested against 7 plants. Galactosamine has been test against 6 plants, with alloxan and lead 5 plants, followed by doxorubicin/Adriamycin and cisplatin 4 plants. Cadmium, aflatoxin B1, Cyclophosphamide, Diclofenac have been tested against 3 plants each while Aluminum chloride, Streptozotocin, 2-acetylaminofluorene (2-AAF), bleomycin (BLM), 7-12, Dimethylbenzanthracene, potassium dichromate/ bromate (K2Cr2O7), Hydralazine, ochratoxin A, Rifampin, dimethyl sulfoxide (DMSO) Petroleum has each been challenged against 2 plant species. Lesser studied toxicants include Bromobenzene, Paraben, phosphamide, carbendazim, malathion, Snake Venom, Pa ge 51 https://journals.e-palli.com/home/index.php/ajmri Am. J. Multidis. Res. Innov. 2(6) 49-56, 2023 Table 1: Plant species tested against the hepatotoxicants Toxicant Plant Species Tested Against References Carbon tetrachloride (CCL4) Azadirachta indica, P. niruri, Phyllanthus lawii, Phyllanthus acidus, Phyllanthus debilis, Vernonia amygdalina, Bidens Pilosa, Taraxacum officinale, Zingibar officinale, Solanum torvum, Syz ygium aromaticum, Allium sativum, Carica papaya, Aloe vera, Khaya senegalensis, Piper guineense, Moringa oleifera, Curcuma longa, Arctium lappa, Brassica rap, Bryonia dioica, Bryophyllum calycinum, Convolvulus arvensis, Caesalpinia crista, Canna indica, Capparis spinosa, Capsella bursa- pastoris, Hyoscyamus Species, Hibiscus rosa-sinensis, Helianthus annuus, Glycyrrhiza glabra, Galium verum, Galium aparine, Fumaria parviflora, Fumaria officinalis, Foeniculum vulgare, Euphorbia hirta, Eupatorium cannabinum, Ephedra foliate, Cuscuta planiflora, Cupressus sempervirens, Coriandrum sativum, Cordia myxa, Clitoria ternatea, Cichorium intybus, Sida acuta, Achillea millefolium, Picrorhiza kurroa, Capparis spinosa, Cicer arietinum, Chenopodium album, Celosia cristata, Zanthoxylum zanthoxyloides Momoh et al., 2018, Bigoniya and Singh, 2014, 2017, Ezzat et al., 2020, Okroro, 2018, Ajiboyea et al., 2018, Ashraf et al., 2018, Chinala et al., 2018, Lee et al., 2017, Enogieru et al., 2015, Abbas et al., 2017, Shyama et al., 2020, Zahra et al., 2012, Akbari et al., 2019, Akbarizare et al., 2021, Ezzat et al., 2020, Lee et al., 2017, Choudhary et al., 2014, Ghareeb et al., 2019, Elmansi et al., 2017, Acheampong et al., 2021 Paracetamol/ Acetaminophen Azadirachta indica, Phyllanthus niruri, Phyllanthus emblica, Phyllanthus niruri, V. amygdalina, Taraxcum officinale, Zingibar officinale, Solanum torvum, Allium sativum, Allium cepa, Moringa oleifera, Momordica charantia, Calotropis procera, Juniperus communis, Hibiscus sabdariffa, Hibiscus cannabinus, Clerodendron inerme, Parkia biglobosa, Saponaria officinalis, Solanum indicum, Maytenus emerginata, Eclipta alba, Aloe vera, Aegle mameloes, Spathodea campanulate, Ficus exasperata Ezzat et al., 2020, Momoh et al., 2015, Usmani et al., 2019, Ige et al., 2017, Iwo et al., 2017, Koubaa et al., 2020, Kermani et al., 2020, Farghali et al., 2015, Mahdi et al., 2019, Momoh et al., 2018, Muhammad et al., 2014, Moracles-Gonzalez, 2014 Ethanol Azadirachta indica, Phyllanthus amarus V. amygalina, Curcuma longa, Phyllanthus niruri, Syz ygium aromaticum, Moringa oleifera, Agrimonia eupatoria, Alhagi maurorum, Anchusa strigose, Zingibar officinale Choudhary et al., 2014, Svenia et al., 2018, Ameaka et al., 2021, Singh et al., 2012, Innih et al., 2022, Maha, 2019, Hashem et al., 2019, Kermani et al., 2020 Thioacetamide Phyllanthus niruri, Taraxacum officinale, Zingibar officinale, Curcuma longa, Allium sativum,Moringa oleifera, Crotalaria juncea Shyama et al., 2020, Sani et al., 2020, Rehab et al., 2016, Sadashiva et al., 2019, Madrigal- Santillan et al., 2014, Chinnala et al., 2018 Galactosamine Phyllanthus maderaspatensis, Bidens Pilosa, Taraxacum officinale, Curcuma longa, Brassica nigra, Cistanche tubulosa Abbas et al., 2017, Zahra et al., 2012, Muhammad et al., 2015, Shyama et al., 2020 Dimethylnitrosamine (DMN) Vernonia amygdalina, Zingiber officinale, Astragalus hamosus, Bauhinia variegate, Cynodon dactylon, Citrullus colocynthis, Spathodea companulata Mahdi et al., 2019, Oyibo et al., 2021, Momoh et al., 2018, Uchenna et al., 2021, Nwaogu et al., 2022 Alloxan Azadirachta indica, Vernonia amygdalina, Curcuma longa, Moringa oleifera, Paullinia pinnata Momoh et al., 2015, Momoh et al., 2018, Koubaa et al., 2020, Salih et al., 2022 Lead Phyllanthus fraternus, Allium sativum, Zingiber officinale, Curcuma longa, Moringa oleifera Kermani et al., 2020, Eghba et al., 2019 Cisplatin Azadirachta indica, Cuminum cyminum, Phyllanthus emblicus, Silymarin Iwo et al., 2017, Tokofai et al., 2021, Ogunmoloye et al., 2022 Cadmium Trema orientalis, Solanum torvum, Syz ygium aromaticum Innih et al., 2022, Olajide et al., 2020 Pa ge 52 https://journals.e-palli.com/home/index.php/ajmri Am. J. Multidis. Res. Innov. 2(6) 49-56, 2023 Aflatoxin B1 Phyllanthus amarus, Calendula officinalis, Parkia biglobosa Farghali et al., 2015, Ajiboye et al., 2014 Cyclophosphamide Phyllanthus fraternus, Hypericum triquetrifoliu, Eucalyptus globulus Usmani et al., 2019, Ghareeb et al., 2019, Ige et al., 2017 Doxorubicin/ Adriamycin Zingiber officinale, Curcuma Longa, Coconut Oil, Vernonia amygdalina Lai et al., 2021, Ezzat et al., 2020, Ige et al., 2017 Diclofenac Curcuma longa, Glycyrrhiza glabra, Moringa oleifera Ashraf et al., 2018, Hassan et al., 2015, Sarfo-Antwi et al., 2018 Streptozotocin Moringa oleifera, Vernonia amygdalina Ajiboyea et al., 2018, Enogieru et al., 2015 Bleomycin (BLM) Curcuma longa, Juglans regia Elzwi, 2019, Yankah et al., 2019 7-12, Dimethylbenzanthracene Turmeric, Garlic Choudhary et al., 2014, Chinnala et al., 2018, Potassium dichromate/ bromate (K2Cr2O7) Moringa oleifera, Allium cepa Elmansi et al., 2017, Akbarizare et al., 2020, Hydralazine Syz ygium aromaticum, Heliotrpium undulatum Patel et al., 2019, Lee et al., 2017 Ochratoxin A Allium sativum, Withania somnifera Jamuna et al., 2018 Rifampin Azadirachta indica, Carthmus tinctorius Akbari et al., 2019, Ezzat et al., 2020 Dimethyl sulfoxide (DMSO) Citrus limon, Citrus aurantifolia Dkhil et al., 2013, Acheampong et al., 2021, Petroleum Vernonia amygdalina, Daucus carota Zahra et al., 2012, Achuba et al., 2020 Eimeria papillate, DMBA, acetylaminofluorene (2-AAF), tert-butyl hydroperoxide (t-BH), PCB, Theobromine, Permethrin, sodium dichromate, nitrobenzene (NB), Diazinon, mercuric chloride, Acrylamide, Methotrexate, N, N-dimethylformamide (DMF) which have been tested against 1 plant species. Table 1 indicates the toxicants and the plant species tested against. Figure 1: Solvents used in extracting phytochemicals tested against toxicants Solvents Used for Extraction Plants In the accepted articles, authors used 11 chemicals to induce hepatotoxicity including Ethanol, Aqueous, Methanol, Phenol, ethylacetate, Petroleum ether, chloroform, hexane, Oil, fermented form, acetone to extract the plant phytochemicals. In 25 toxicants used to induce liver diseases, ethanol was the solvent used for extracting the plant parts while methanol was the solvent in 16 toxicants tested and distilled water (13 toxicants) as indicated in figure 1. In-Vitro and In-Vivo Models Liver Toxicity were Induced with the Toxicants Hepatotoxicity was induced in 10 in-vitro and in-vivo models including Wister Albino rats, Sprague-Dawley rats, HepG2 cell culture line, mice, rabbits, clone-9 cells, broiler chicken, goat hepatocytes, guinea pigs, Swiss albino rats and then tested against the extracted plant phytochemicals. Hepatoxicity was induce in Wistar albino rats using 46 different toxicants followed by Sprague- Dawley rats and Hep G2 cell cultures with 6 different toxicants each. Toxicity of hepatocytes was also reported in mice models using 4 different toxicants while 3 separate toxicants were used to induced liver diseases in Swiss albino rats. Rabbits, clone-9 cells, broiler chicken, Goat Pa ge 53 https://journals.e-palli.com/home/index.php/ajmri Am. J. Multidis. Res. Innov. 2(6) 49-56, 2023 hepatocytes, Guinea pigs were induced with liver diseases using a single toxicant each as indicated in Figure 2. DISCUSSION The review indicates that CCL4 is most prevalent toxicant 55/94 plant species (58.5%) with ethanol being the most solvents 25/55 toxicants (45.5%) and Wistar albino rats being the commonest 46/55 toxicants (83.6%) in-vivo hepatotoxicity inducing model. Despite being a known respiratory inhibitor as it binds and oxidise heme in haemoglobin (Talluri et al., 2018), CCL4 now banned is one of earliest discovered toxicant (Enogieru et al., 2015; Innih et al., 2022). CCL4 is less expensive and can be obtained easily for study purposes. Due to its availability in the environment from fridges and industrial chemicals, CCL4 hepatotoxicity also represent a real-time risk of liver diseases. Phytochemical compounds are organic and therefore dissolve on the principle of polar substance dissolve in polar solvents (Senti et al., 2016). Comparatively, ethanol comes next to phenol in terms of polarity and will therefore yield high phytochemicals. Attenuating hepatotoxicity induced by oxidative stress has been dose-dependent (Oboma et al., 2018, Momoh et al., 2018). Therefore, the higher the phytochemical yield, the more effective oxidative stress remediation. In a comparative study, Mahdi et al. 2019, reported that hydro-ethanolic had better hepatotoxicity than same concentration of methanol, chloroform and ether account of solvent polarity. It is also reported that, moderate ethanol concentrations 50-70% had better hepatoprotective activity that lesser concentration and higher concentrations (Shah et al., 2016). At lower concentration, phytochemical yield is less while at higher concentration, cellular parts and toxic antinutrients are extracted which pose further hepatic harm. Wistar albino rats are more adaptable to environmental conditions with less case fatalities during experimentation. They rapidly progenerate making them less expensive and available for research purposes (Ajiboyea et al., 2018). This adaptability to human weather conditions also make it suitable to real human hepatic conditions. CONCLUSION Many toxicants have been reported to induce liver diseases in various models, however, several medicinal plants have been reported to ameliorate these induced hepatic diseases. Medicinal plants therefore present hope for the discovery of ideal hepatorestorative agents that will be less expensive largely due to their ability to dissolve in common solvents like ethanol. REFERENCES Abbas, N., Naz, M., Alyousef, L., Ahmed, E. S., Begum, A. (2017). Comparative study of hepatoprotective effect produced by cuminum cyminum, fruits of Phyllanthus emblicus and Silymarin against cisplatin- induced hepatotoxicity. International Journal of Pharmaceutical Sciences and Research, 8(5), 2026-2032. Acheampong, D. O., Baffour, I. K., Atsu Barku, V. Y., Addo, J. K., Essuman, M. A., & Boye, A. (2021). Zanthoxylum zanthoxyloides alkaloidal extract improves CCl4- induced hepatocellular carcinoma-like phenotypes in rats. Evidence-Based Complementary and Alternative Medicine, 2021. https://doi.org/10.1155/2021/3804379 Achuba, F. I. and Ichipi-Ifukar, P. C. (2019). Protective effect of Vernonia amygdalina methanolic extract against hepatocellular damage induced by petroleum contaminated diet in male rats. Iraq Journal of Science, 61(11), 2820-2830. Ajiboyea,T. O., Adeleyeb, A. O., Salaub, A. K., Ojewuyia,O.B., Adigunc, N.S., Saheed S., Taofik, O. S. (2018). Phenolic extract of Parkia biglobosa fruit pulp stalls aflatoxin B1 – mediated oxidative rout in the liver of male rats. Brazilian Journal of Pharmacognosy, 24, 668-676. Akbari, A., Nasiri,K., Heydari,M., Nimrouzi, M., Afsar, T. (2019). Ameliorating Potential of Ginger (Zingiber officinale Roscoe) Extract on Liver Function and Oxidative Stress Induced by Ethanol in Male Rats. Zahedan Journal of Research and Medical Science, 21(2), e86464. Akbarizare, M., Ofoghi, H., Hadizadeh, M., Moazami, N. (2020). In vitro assessment of the cytotoxic effects of secondary metabolites from Spirulina platensis on hepatocellular carcinoma. Egyptian Liver Journal, 10, 11-21. Figure 2: In-vitro and in-vivo models used to model hepatotoxicity Pa ge 54 https://journals.e-palli.com/home/index.php/ajmri Am. J. Multidis. Res. Innov. 2(6) 49-56, 2023 Akharaiyi, A. C., Okafor, A. C. (2021). Effect of Spathodea campanulata Ethanol Leaf Extract on Hematology and Liver Function of Salmonella infected and Paracetamol-induced Swiss Albino Mice. FABAD Journal of Pharmaceutical Science, 46(3), 251-260. Akpanyung, E. O., Bassey, U. E., Udofia, E. K., Effiong, G. S. (2019). Effect of Ethanol Leaf Extract of Vernonia amygdalina on Some Indices of Liver, Kidney Function and Lipid Profile in Theobromine Intoxicated Male Albino Wistar Rats. Journal of Food and Nutrition Sciences, 6(4), 106-114. Al-Snafi, A. E., Mousa, H. N., Majid, W. J. (2019). Medicinal plants possessed hepatoprotective activity. IOSR Journal of Pharmacy, 9(8), 26-56. Aly, O., Abouelfad, D. M., Shaker, O. G., Hegazy, G. A., Fayez, A.M., Zaki, H.H. (2020). Hepatoprotective effect of Moringa oleifera extract on TNF-α and TGF-β expression in acetaminophen-induced liver fibrosis in rats. Egyptian Journal of Medical Human Genetics, 21, 69-78. Ameaka, F. N., Tembe, E., Tchadji, M. V. E., Bayaga, H., Dobgima, J. F., Eustace, B. B., Njinkio, B. N., Tabi, Y. O., Ngameni, B., Fokunang, C. (2021). Phytochemical Characterization, Hepatoprotective Activity on Alcohol-Induced Toxicity of the Aqueous Extract of Curcuma longa (Zingiberaceae) in Wistar Rats. Journal of Complementary and Alternative Medical Research, 16(4), 134-149. Ashraf, E., Aboubakr, M., Ibrahim, S., Abdelhamid, Y. (2018). Protective effect f Szygium aromaticum (clve) oil against acrylamide-induced hepatic, renal and testicular damage in albino rats. International journal of Pharmacology and Toxicology, 6(1), 12-17. Bigoniah, P. and Singh, C. S. (2014). Hepatoprotective activity of a standardized poly-herbal liver formulation. International Journal of Pharmaceutical Science and Research, 5(10), 4209-4218. Chinnala, K. M., Jayagar, P. P., Motta, G., Adusumilli, R. C., Elsani, M. M. (2018). Evaluation of hepatoprotective activity of Allium sativum ethanolic extract in thioacetamide-induced hepatotoxicity in albino Wistar rats. American Journal of Research in Medical Sciences, 3(2), 48–53. Choudhary, V., Bibin, B. A., Lahkar, M. (2014). Hepatoprotective effect of Azadirachta indica in alcohol induced liver damage. World Journal of Pharmaceutical Research, 3(4), 1913-1925. Dkhil, M. A., Al-Quraishy, S., Aref, A. M., Othman, M. S., El-Deib, K. M., & Abdel Moneim, A. E. (2013). The potential role of Azadirachta indica treatment on cisplatin-induced hepatotoxicity and oxidative stress in female rats. Oxidative Medicine and Cellular Longevity, 2013. http://dx.doi.org/10.1155/2013/741817. Eghba, M. A., Anoush, M., Ghoreyshi, A., Heidari, R.(2019). The Cytoprotective Effects of Allium cepa Methanolic Extract in Freshly Isolated Hepatocytes. Trends in Pharmaceutical Sciences, 5(4), 207-216. Elmansi, A. M., El-Karef, A. A., El-Shishtawy, M. M., Eissa, L. A. (2017). Hepatoprotective Effect of Curcumin on Hepatocellular Carcinoma Through Autophagic and Apoptic Pathways. Annals of Hepatology, 16(4), 607-618. Elzwi, S. (2019). Effect of Zingiber officinale (Ginger) Extract on Acetaminophen- Induced Hepatotoxicity in Mice. Pharmaceutical Methods, 10(1), 27-30. Enogieru, A. B; Charles, Y. O; Omoruyi, S. I.; Momodu, O. I; Ezeuko, V. C. (2015). Stem Bark Extracts of Ficus exasperata protects the Liver against Paracetamol induced toxicity in Wistar Rats. Journal of Applied Science and Environmental Management, 19(1), 155-159. Ezzat, M. I., Okba, M. M., Ahmed, S. H., El-Banna, H. A., Prince, A., Mohamed, S. O. (2020). In-depth hepatoprotective mechanistic study of Phyllanthus niruri: In vitro and in vivo studies and its chemical characterization. PLoS ONE, 15(1), e0226185 https://doi.org/10.1371/journal.pone.0226185 Farghali, H., Canová, N. K., Zakhari, S. (2015). Hepatoprotective properties of extensively studied medicinal plant active constituents: Possible common mechanisms. Pharmaceutical Biology, 53(6), 781-791, https://doi.org/10.3109/13880209.2014.950387. Ghareeb, M. A., Sobeh, M., El-Maadawy, W. H., Mohammed,W. S., Khalil, H., Botros, S., Wink, M. (2019). Chemical Profiling of Polyphenolics in Eucalyptus globulus and Evaluation of Its Hepato– Renal Protective Potential Against Cyclophosphamide Induced Toxicity in Mice. Antioxidants, 8, 415; https:// doi.org/10.3390/antiox8090415. Hashem, M. M, Salama, M. M., Mohammed, F. F., Tohamy, A. F., El Deeb, K. S. (2019). Metabolic profile and hepatoprotective effect of Aeschynomene elaphroxylon (Guill. & Perr.). PLoS ONE, 14(1), e0210576.https:// doi.org/10.1371/journal. pone.0210576. Hassan F., Nikolina K. C., Samir Z. (2015). Hepatoprotective properties of extensively studied medicinal plant active constituents: Possible common mechanisms. Pharmaceutical Biology, 53, 6, 781-791, https://doi.org/10.3109/13880209.2014.950387. Ige, S. F., Adeniyi, J. M., Iyalla, G. O. (2017). Allium cepa Mitigates Aluminum Chloride-Induced Hepatotoxicity in Male Wistar Rats. Journal of Biomedical Sciences, 6(4), 27-37. Innih, S. O., Agu, K. C., Eze, I. G. (2022). Immunomodulatory and hepatoprotective properties of Solanum torvum (Turkey berry). Sahel Medical Journal, 21(1), 154-160. Iwo, M. I., Sjahlim, S. L., Rahmawati, F. S. (2017). Effect of Vernonia amygdalina Del. Leaf Ethanolic Extract on Intoxicated Male Wistar Rats Liver. Scientia Pharmactica, 85, 16. https://doi.org/10.3390/ scipharm85020016 Jamuna, G., Sharma, A. K., Manimaran, A., and Palanisamy Sankar, P. (2018). Hepatoprotective effects of Allium sativum and Withania somnifera on ochratoxin A-induced toxicity in rats. Journal of Pharmacognosy and Phytochemistry, 7(3), 2675-2680. Pa ge 55 https://journals.e-palli.com/home/index.php/ajmri Am. J. Multidis. Res. Innov. 2(6) 49-56, 2023 Kermani, N. M., Aldaek, A. M., Abushofa, F. A, Jaat, F. G. (2020). Protective Effect of Allium cepa L.(onion) Against Potassium Bromate-Induced Hematological, Biochemical and Histopathological Alterations in Rats. International Journal of Innovative Science and Research Technology, 5(11), 201-211. Koubaa, F. G., Chaâbane, M., Choura, B., Mouna Turki,M., Makni-Ayadi,F., El Feki, A.(2020). Hepatoprotective Effects of Taraxacum officinale Root Extract on Permethrin-induced Liver Toxicity in Adult Mice. Pharmaceutical and Biomedical Research, 6(3), 223-236. http://dx.doi.org/10.18502/ pbr. v6i3.4649. Lai, C-C., Zhou, X., Wang, H-K., Lin, Y-C., Lin, H-Y., Wang, T-D., Liu, B-L. (2021). Vernonia amygdalina extract induces apoptosis and inhibit epithelial- mesenchymal transition in hep 3B cells through the inhibition of P13K/Akt signaling pathway. International Journal of Applied Science and Engineering, 19(1), 86-95. Lee, C-H., Lee, H-Y., Chung, H-Y., Kim, H-W., Chae, H-J. (2017). Protective effect of Curcuma longa extract on CCL4-induced acute hepatic stress. BMC Research Notes, 10,77. Madrigal-Santillan, E., Madrigal-Bujaider, E., Alveraz- Gonalez, I., Sumaya-Martinez, M.T., Gutierrez- Salinas, J., Bautista, M., Morales-Gonzalez, A., Garcia-Luna, M, G-R., Aguilar-Faisal, Moracles- Gonzalez, J.A. (2014). Review of some natural plants with hepatoprotective effects. World Journal of Gastroenterology, 20(40), 14787-14807. Maha, A. A. (2019). Evaluation of Hepatoprotective Activity of Neem Extract in Rifampin Induced Acute Hepatic Failure in Rats. International Journal of Pharmaceutical Research & Allied Sciences, 8(3), 29-36. Mahdi, M. T., Mawahib, G. A., Karima, F. A. (2019). Hepatoprotective and nephroprotective effects of the aqueous extract of turmeric (curcuma longa) in rifampicin and isoniazid-induced hepatotoxicity and nephrotoxicity in rats. Asian Journal of Pharmaceutical and Clinical Research, 12(3), 293-298. Mirazi N., Karami, Z. (2016). The protective effect of hydroalcoholic extract from rhizome of Zingiber officinale L. on carbon tetrachloride-induced hepatic injury in male rat. Journal of Kashan University of Medical Sciences, 20(4), 297-305. Mohamed A. L., Nabil, M. T., Mahdy, A. K., Abd El- Wahab A. M., and Raghda, I. G. (2020). Ginger (Zingiber officinale) potentiate paracetamol induced chronic hepatotoxicity in Rats. Journal of Medicinal Plants Research, 7(42), 3164-3170. Momoh, J., Long, O, O., Damazio, O. A., Eleyowo, O. O. (2015). Hepatoprotective Effect of Ethanolic Leaf Extract of Vernonia amygdalina and Azadirachta indica against Acetaminophen-Induced Hepatotoxicity in Sprague-Dawley Male Albino Rats. American Journal of Pharmacological Sciences, 3(3), 79-86. Momoh, O. J., Olaniyi, A. M., Aderele, O. R. (2018). Experimental and Mathematical model for the hepatoprotective effect of methanolic extract of Moringa oleifera in Sprague-Dawley albino rats. Journal of Advances in Medicine and Medical Research, 26(5), 1-14. Muhammad, I. V., Alhassan, A. J., Wudil, A. M., Jarummi, I. K. (2015). Toxicological and protective effects of aqueous stem bark of Khaya senegalensis (ASBEKS) on liver of experimental rats. British Journal of Applied Science and Technology, 6, 600-605. Nwaogu, J., Sanusi, M., Ahmed Jega, A., Anka, S.S.(2022). Hepatoprotective Effect of Parkia Biglobosa Husk Methanol Extract Against Carbon Tetrachloride (CCL4) Induced Liver Damage in Albino Rats. International Journal of Innovative Science and Research Technology, 7(2), 1073-1083. Oboma, Y., Beredugu, S., Okara, P. N., Tumuno-Omie, F. A., Ibiang, O. E. (2018). Protective effect of combined extracts of Allium sativum and Zingibar officinale against lead acetate induced hepatotoxicity and testicular damage in Rattus norvegitus. MOJ Anatomy and Physiology, 5(5), 45-55. Ogunmoloye, T., Falusi, O. O., Oderinde, F. (2022). Sida acuta leaf extract attenuates oxidants-induced model of nephrotoxicity and hepatotoxicity. Clinical Phytoscience, 8, 5. Okaiyeto K, Nwodo UU, Mabinya LV, Okoh AI. (2018). A review on some medicinal plants with hepatoprotective effects. Phamacognosy Reviews, 12, 186-199. Okoro, I. O. (2020). Hepatoprotective effect of hydroethanolic extracts of Allium sativum and Carica papaya on CCL-4 induced liver damage in rats. Nigerian Journal of Pure and Applied Sciences, 33, 3639- 3645. Olajide, J. E., Sanni, M., Achimugu, J. O., Suleiman, M. S., Jegede , E. R., Sheneni, V. D.(2020). Effect of methanol extract of Trema orientalis leaf on some biochemical and histopathological indices of wistar albino rats with cadmium-induced hepatotoxicity. Scientific African, 10 (2020), e00568 Olatosin T. M., Akinduko D. S., Uche C. Z., Bardi J. (2014). Effects of Moringa oleifera Seed Oil on Acetaminophen-Induced Oxidative Stress and Liver Damage in Wistar Albino Rats. IOSR Journal of Pharmacy and Biological Sciences, 9(2), 53-59. Onoja, S. O., Daniel-Igwe, G., Ejiofor, E., Mbakwe, C. C., Ugochuhwu, S. O., Maxwell I. Ezeja, M., Yusuf, N. O, Asuzu, I.U.(2019). Hypolipidemic, hepatoprotective, nephroprotective and anti-lipid peroxidation properties of a methanol extract of Paullinia pinnata root-bark, in alloxan-induced hyperglycemic rats. Current Issues in Pharmacy and Medical Sciences, 32(3), 125-129. Oyibo, A., Gbadegesin, M. A., Odunola, O. A. (2021). Ethanol extract of Vitellaria paradoxa (Gaertn, F) leaves protects against sodium arsenite - induced toxicity in male Wistar rats. Toxicology Reports, 8, 771-778. Pa ge 56 https://journals.e-palli.com/home/index.php/ajmri Am. J. Multidis. Res. Innov. 2(6) 49-56, 2023 Patel, U. D., Shah, F. C., Jain, N. K. (2019). Hepatoprotective effect of methanolic extract of Syzygium aromaticum against hydralazine induced toxicity: An In vitro study. The Pharma Innovation Journal, 8(6), 537-541. Ramírez-Marroquín, O. A., Jiménez-Arellanes, M. A. (2019). Hepato-Protective Effect from Natural Compounds, Biological Products and Medicinal Plant Extracts on Antitubercular Drug-Induced Liver Injuries: A Systematic Review. Medicinal & Aromatic Plants, 8(5), 339. Rehab, A. S., Magdy, M, M., Mahmoud, B. A., Marwa M. S. (2016). Assessment effect of Aloe vera, Azadirachta indica and Moringa oleifera aqueous extracts on carbon tetrachloride-induced hepatotoxicity in rats. International Journal of Pharmacy and Pharmaceutical Sciences, 8(4), 83-89. Sadashiva, C. T., Hussain, F. H. M., Nanjundaiah, S. (2019). Evaluation of hepatoprotective, antioxidant and cytotoxic properties of aqueous extract of turmeric rhizome (Turmesac®). Journal of Medicinal Plants Research, 13(17), 423-430. Salih, A. I., Saleh, M. H., Khalaf, S., Ayed, S. (2022). Effect of Moringa oleifera Leaves against Hepatotoxicity Induced by Bisphenol A. Archives of Razi Institute, 77(3), 1083-1089. Sani, I., Umar, R. A., Hassan, S. W., Faruq, U. Z., Bello, F., Aminu,H., Sulaiman, A. (2020). Hepatoprotective Effect of Azadirachta indica Leaf Fractionated Extracts against Snake Venom Toxicity on Albino Rats. Saudi Journal of Biomedical Research, 5(6), 112-117. Sarfo-Antwi, F. S., Larbie, C., Babatunde, D. (2018). Extracts of Ageratum Conyzoides L. Protects against Carbon Tetrachloride – Induced Toxicity in Rats through Inhibiting Oxidative Stress. Journal of Advances in Medical and Pharmaceutical Sciences, 19(2), 1-14. Sentí M, Tomás M, Fitó M, Weinbrenner T, Covas M, Sala J, Masiá R, Marrugat J. (2016). Antioxidant paraoxonase 1 activity in the metabolic syndrome. Journal of Clinical Endocrinology & Metabolism, 88, 5422- 5426. Shah, F. C. and Jain, N. K. (2016). In Vitro study on hepatoprotective effect of Phyllanthus fraternus against lead induced toxicity. UK Journal of Pharmaceutical and Biosciences, 4(2), 31-37. Shyama, K. D., Soumendra, N. K., Sampurna R. (2020). Protective action of hydroethanolic extract of Moringa oleifera flower on acetaminophen-induced hepatotoxicity in rats. International Journal of Green Pharmacy, 14(1), 98-108. Singh, I., Vetriselvan, S., Shankar, J., Gayathiri1, S., Hemah, C.,Yaashini, I. G. A. (2012). Hepatoprotective activity of aqueous extract of Curcuma longa in ethanol induced hepatotoxicity in albino Wistar rats. International Journal of Phytopharmacology, 3(3), 226-233 Svenia, P. J., Ratheesh, M., Asha, S., Krishnakumar, I. M., Sandya, S., Kumar, B. G. (2018). Hepato-protective Effect of Clove Bud Polyphenols (Syzygium aromaticum L.) (Clovinol by Modulating Alcohol Induced Oxidative Stress and Inflammation. Journal of Food Research, 7(1), 10-20. Talluri, R. M., Gummadi, V. P., Battu, R. G. (2018). Chemical composition and hepatoprotective activity of Saponaria officinalis on paracetamol-induced liver toxicity in rats. Pharmacognosy Journal, 10(6), 1196-1201. Tokofai, B. M.; Idoh, K.; Oke, O. E.; Agbonon, A. (2021). Hepatoprotective Effects of Vernonia amygdalina (Astereaceae) Extract on CCl4-Induced Liver Injury in Broiler Chickens. Animals, 11, 3371-3381. https:// doi.org/10.3390/ani11123371. Uchenna, O. M., Nwozo, S. O., Mohammed, B. A., Garba, R., Mohammed, Y. A. (2021). Spathodea campanulate (Africa Tullip tree) stem and root barks extracts ameliorate N-nitrosodiethylamine induced hepatic impairment in male rats. Biomed Natural and Applied Sciences, 1(1), 57-65. Usmani, S., Qureshi,H. J., Zaheer, A.(2019). Hepatoprotective and antioxidative effects of Allium sativum var lehsun gulabi on acetaminophen induced acute hepatitis in male albino rats. Pakistani Journal of Physiology, 15(1), 32–36. Usunomena, U., Okolie N., Eze I. G. (2015). Inhibitory Effect of Vernonia amygdalina on Dimethylnitrosamine (DMN)induced Liver Fibrosis in Rats. International Journal of Clinical Pharmacology and Toxicology, 4(4), 179-184. Zahra, K., Malik, M. A., Mughal, M. S, Arshad, M., Sohail, M. I.(2012). Hepatoprotective role of extracts of Momordica charantia in acetaminophen-induced toxicity in rabbits. The Journal of Animal and Plant Sciences, 22(2), 273-277.