Neuroprotective Efficacy of Lycopene in a Thioacetamide-Induced Model of Hepatic Encephalopathy: An Experimental Rat Study

Neuroprotective Efficacy of Lycopene in a Thioacetamide-Induced Model of Hepatic Encephalopathy: An Experimental Rat Study | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Neuroprotective Efficacy of Lycopene in a Thioacetamide-Induced Model of Hepatic Encephalopathy: An Experimental Rat Study Erdal Tekin, Ersan Gursoy, Fatma Tortum, Mehmet Nuri Koçak, Ufuk Okkay, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6819328/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Objective Lycopene has known antioxidant, anti-inflammatory, anti-proliferative, and neuroprotective effects. This is the first study to evaluate the potential protective role of lycopene against hepatic encephalopathy. Methods An experimental hepatic encephalopathy model was established by intraperitoneal administration of thioacetamide to rats. A total of 24 rats were randomized into four groups: healthy group, control group, low-dose (50 mg/kg) lycopene group, and high-dose (100 mg/kg) lycopene group. The locomotor activity test was measured on the first day of the study to determine baseline measurements and again at the end of the study to determine changes. Blood samples were collected, and liver, brain, and lungs were removed and weighed. Results Except for stereotypic movements (p > 0.05), all other final locomotor activity tests were statistically significant (p 0.05), but IL-6, IL-10 and IL-1ꞵ analysis results were significant between control and healthy and high-dose lycopene groups (p < 0.05). AST, ALT, total bilirubin, direct bilirubin and ammonia parameters were significantly lower in the low-dose lycopene group compared to the control and high-dose lycopene groups (p < 0.05). Conclusion Lycopene administration had hepatoprotective and neuroprotective efficacy against HE, supported both in locomotor activity tests and at biochemical tests. Experimental study hepatic encephalopathy lycopene neuroprotection Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 1. INTRODUCTION Lycopene is a powerful antioxidant that belongs to the carotenoid family and is found in red fruits and vegetables. Lycopene can be used in the treatment of many diseases and studies have shown that it has antioxidant, anti-inflammatory, anti-proliferative, anti-cancer and other properties [ 1 , 2 ]. Lycopene is hepatoprotective against several liver diseases, including alcoholic liver disease, nonalcoholic fatty liver disease, hepatic fibrosis, ischemia/reperfusion injury, hepatocellular carcinoma, fulminant liver failure and radiation injury [ 3 – 5 ]. Lycopene is thought to be a potent neuroprotectant against oxidative stress and could be used to prevent the injury or death of nerve cells in a number of neurological diseases [ 6 ]. Animal studies have highlighted its ability to adequately cross the blood-brain barrier and to improve many neurodegenerative disorders [ 7 ], and another study found that lycopene was even neuroprotective against hypoxia-ischemic injury in rats [ 8 ]. Hepatic encephalopathy (HE) is a common complication of acute and chronic liver disease and is accompanied by neuropsychiatric disturbances. The pathogenesis of HE is multifactorial and has not yet been fully elucidated [ 9 ]. However, morphological changes in astrocytes are thought to be caused by neurotoxic substances such as ammonium and manganese. This leads to brain dysfunction, brain atrophy and brain edema as a result of disruption of the blood-brain barrier [ 10 – 12 ]. These metabolic abnormalities of HE are considered reversible. However, most patients are diagnosed after reversible brain damage has occurred, at which time neuropsychiatric disorders develop [ 12 ]. As HE has a high morbidity and mortality, new aspects need to be addressed to improve both liver and neurological disease. Lycopene may have improved HE because of its known antioxidant, anti-inflammatory, anti-proliferative and neuroprotective properties. This study aimed to evaluate the potential protective role of lycopene against HE and behavioral consequences in a rat model of thioacetamide-induced HE. 2. MATERIAL AND METHOD 2.1. Study Design This study is an experimental study and the ethics committee approval for animal experiments was obtained from the Local Ethics Committee for Animal Experiments of our university (Decision No: 28.01.2021- 2021/1–25). The handling of rats and experimental procedures were performed according to the guidelines for experimental animals. 2.2. Animals A total of 24 healthy albino male Sprague-Dawley rats weighing 250 ± 25 g were used in the study. The rats were obtained from the Medical Experimental Application and Research Center of our university. The animals were housed in groups under controlled lighting conditions (12:12 h light/dark cycle) in a well-ventilated room at 22 ± 1°C in typical plastic cages on sawdust beds. Rats were fed ad libitum throughout the study. 2.3. Materials An experimental HE model was established by intraperitoneal administration of thioacetamide (TAA) to rats. TAA is a sulfur-containing catalyst that causes liver fibrosis and hepatotoxicity by inducing activation of macrophages and secretion of various inflammatory cytokines [ 13 ]. Studies in the literature have found TAA to be a good agent in an experimentally induced HE model in rats. [ 14 – 16 ]. TAA (Sigma-Aldrich, St. Louis, MO, USA) was prepared fresh by dissolving in sterile distilled water. Lycopene 10% FS (Redivivo, DSM Nutritional Products) was prepared by dissolution in corn oil and administered to the rats by gavage. 2.4. Study Groups A total of 24 rats were randomized into four groups of six rats each: healthy group, control group, low dose lycopene group and high dose lycopene group. Group 1: Healthy group (n = 6); no treatment was administered and only weight was monitored. Group 2: Control group (n = 6); weight was monitored for 15 days and 200 mg/kg TAA was administered intraperitoneally for 3 consecutive days from day 15 to day 17. On the days of TAA administration, fluid replacement was performed with lactated ringer (10 mg/kg/day) in the morning and evening. Group 3: Low-dose lycopene group (50 mg/kg, n = 6); weight was monitored for 15 days and lycopene 50 mg/kg was administered by gavage at the same time every day until the 15th day of the experiment. From the 15th to the 17th day of the experiment, 200 mg/kg TAA was administered intraperitoneally every day for 3 consecutive days. On the days of TAA administration, fluid replacement with lactated ringer (10 mg/kg/day) was performed in the morning and evening. Group 4: High-dose lycopene group (100 mg/kg, n = 6); body weights were monitored for 15 days and lycopene 100 mg/kg was administered by gavage at the same time every day until the 15th day. From the 15th to the 17th day of the experiment, 200 mg/kg TAA was administered intraperitoneally every day for 3 consecutive days. On the days of TAA administration, fluid replacement was performed in the morning and evening with lactated ringer (10 mg/kg/day). 2.5. Locomotor Activity Test The locomotor activity test was measured with an open-field activity imager system (May Act 508) on the first day of the study to determine baseline measurements and again on day 18 at the end of the study to determine changes. The basis of the system used to measure locomotor activity is a square quadrilateral with infrared light sources on each side. A square Plexiglas cage is placed inside this quadrilateral. When the animal makes a movement inside the cage, it interrupts the communication between the opposite infrared sensors and this is recorded by a recorder connected to the device according to the shape of the movement made by the subject. With the help of this system, horizontal, vertical and ambulatory activities of rats are recorded. Horizontal movement is the movement of the experimental animal in place without any displacement or standing movement. Vertical movement is a standing movement and is detected by vertical sensors on the bands. Ambulatory movement is any displacement movement of the animal in the cage other than standing. Horizontal and vertical activities give an idea of the animal's stereotypical movements and aggressiveness. All three activities can be evaluated separately or the sum of the three can be expressed as total locomotor activity. Resting times and distance traveled were measured and recorded. All rats were placed individually in a transparent cage twice, once at the beginning and once at the end of the study, and recorded for 10 minutes to determine whether locomotor activity decreased or increased. The flow chart of the study is shown in Fig. 1 . 2.6. Study Protocol The same preoperative preparation, anesthesia and surgical technique were used for each group. All rats were euthanized under ketamine/xylazine anesthesia when the study was completed. Blood samples were collected for biochemical and hematological analysis. Liver, brain and lungs of all rats were removed, weighed and compared to investigate possible infection and inflammation in the organs. 2.7. Biochemical analysis Blood samples (5 cc) were collected from all rats, clotted at room temperature, then placed in a yellow-capped gel tube and centrifuged at 3000 rpm for 10 minutes. Serum samples were placed in Eppendorf tubes and analyzed for biochemical parameters. Additionally, blood samples of 3cc were collected from all rats and placed in EDTA-containing hemogram tubes according to the cold chain rules, and ammonium analysis was performed immediately after plasma separation. AU 5800 Beckman coulter autoanalyzer® (USA) and standard kits were used to measure serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin, direct bilirubin and plasma ammonium levels. Bioassay Technology Laboratory (BT LAB®, Shanghai, Korain) rat kits were used for ELISA (enzyme-linked immunosorbent assay) analysis of interleukin (IL) 1beta, IL-1RA, IL-4, IL-6 and IL-10 levels. 2.8. Statistical Analysis In our study, statistical analyses were performed with SPSS 25.0 (IBM Corp., NY, USA). Categorical data were presented as percentage and frequency and numerical data as mean and standard deviation. Kolmogorov-Smirnov test was used for normal distribution evaluation. Mann Whitney U test and Kruskal Wallis variance test were used for statistical analysis between groups. Dunnett T3 was used for post-hoc analysis of the data. Statistical significance was taken as p < 0.05 in the whole study. 3. RESULTS 3.1. Weights of the rat and its organs The mean weights of all groups that were weighed every three days within the scope of our study are shown in Fig. 2 a. The mean weights of the rats included in the study were 277.67 ± 9.32 and there was no statistically significant difference (p = 0.172). After euthanasia, the organs of the rats were removed and the weighed organ weights are shown in Fig. 2 b. Accordingly, there was no significant difference in organ weights between groups (lung, liver and brain p values were 0.254, 0.371, 0.903, respectively). 3.2. Locomotor activity test results Locomotor activity tests were performed in all rats included in the study both at the beginning and at the end of the study. Basal locomotor tests are given in Fig. 3 . Accordingly, when the control group was compared with the other groups, stereotypic movement and resting time duration were higher in the control group, while other locomotor activity movements were lower. While there was no statistical significance between the groups in stereotypic, ambulatory and vertical locomotor activity tests (p > 0.05), there was statistical significance in other locomotor activities (p 0.05), other locomotor activity tests were statistically significant (p < 0.05). Ambulatory, vertical, and horizontal movement were higher in healthy rats and the healthy group was statistically significant compared to the other groups (p < 0.05). Resting time was less in healthy rats and statistically significant (p < 0.05). 3.3. Biochemical analysis results The results of interleukin analysis in the blood samples of the rats taken at the end of the study are given in Fig. 5 . Accordingly, there was no statistical significance in IL-4 and IL-1RA analysis results (p > 0.05). IL-6, IL-10 and IL-1ꞵ analysis results were significant and as a result of the post-hoc analysis, there was statistical significance between the control group and the healthy and high dose lycopene group (p < 0.05). Biochemical analysis results of the blood samples of the rats taken at the end of the study are given in Fig. 6 . Accordingly, there was statistical significance in the analysis results (p < 0.05). It was determined that AST, ALT, total bilirubin, direct bilirubin and ammonia parameters were lower in the low dose lycopene group compared to the control and high dose lycopene group, which was statistically significant (p < 0.05). 4. DISCUSSION In this study, weight and organ weights of HE-induced rats were calculated and locomotor activity tests and biochemical analyses were performed for behavioral defects. As a result of this study, it was determined that the weight of weight increased in the low dose lycopene group, while liver weight increased in the control group. Locomotor activity tests generally decreased but total distance traveled increased in both groups treated with lycopene. In addition, it was determined that IL-6, IL10, IL-1Beta, ALT, AST and ammonia levels decreased in both groups treated with lycopene compared to the control group. These results showed that lycopene has a protective effect against THA-induced HE. In HE, excessive amounts of ammonia cross the blood-brain barrier and disrupt brain metabolism and astrocyte structure, leading to cognitive and motor symptoms. With hyperammonemia, neuroinflammation occurs, contributing to intellectual disability [ 10 , 17 , 18 ]. Lycopene has the ability to counteract neuroinflammation, proteinopathies, apoptosis, synaptic dysfunction and brain edema, as well as ameliorating oxidative stress. In the literature, these neuroprotective properties of lycopene have been emphasized to ameliorate cognitive deficits, locomotor disorders, dementia, seizures, anxiety and behavioral abnormalities including depression-like behaviors associated with neurological disorders [ 19 , 20 ]. In our study, there was deterioration in the locomotor activity tests of the other groups compared to the healthy group in animal models of HE. Locomotor activity test results in both groups treated with lycopene were better than the control group. This shows that lycopene improves locomotor activity tests in HE. In addition, in our study, ammonia levels were found to be lower in the lycopene -treated groups compared to the untreated group. This shows us that leukopenia can treat high ammonia levels in HE. Thus, we think that lycopene can be used in the treatment of cognitive and cognitive functions caused by high ammonia levels. HE occurs as a result of the interaction of many pathophysiologic factors in brain metabolism such as damage in blood-brain barrier permeability, inflammation and oxidative stress [ 18 , 21 , 22 ]. Most of the drugs used to treat HE also aim to reduce blood serum ammonia and suppress neurotoxin production [ 10 , 23 ]. In addition, inflammation and oxidative stress accelerate the progression of HE [ 24 ]. Antioxidant and anti-inflammatory drugs have been reported to have protective effects on the brain and liver [ 25 , 26 ]. There are no studies investigating the efficacy of lycopene, which has antioxidant and anti-inflammatory properties, in HE. In this study, we aimed to investigate this efficacy and this is the first study in the literature. As the results of our study showed that locomotor activity test results improved, it was concluded that lycopene improves cognitive function damage in HE. In the literature, it has been emphasized that lycopene has hepatoprotective properties against liver diseases and improves memory and cognition abilities of rodents in animal experiments [ 4 , 7 ]. In the HE model; it was determined that lycopene administered orally for 15 days had hepatoprotective and cognitive enhancing effect in THA-induced HE and this is the first study showing this effect. In the literature, there are studies reporting that liver enzyme levels of rats are impaired in THA-induced HE. In these studies, it was emphasized that ALT and AST levels increased and ammonia concentration increased. These studies show the severity of liver damage [ 27 – 31 ]. Similar to the studies in the literature, hepatic enzyme activity and ammonia level increased in our study. In addition, liver damage was attenuated with leukopenia treatment. Cognitive and neurological impairments detected in HE models were associated with increased ammonia levels, oxidative stress and pro-inflammatory cytokines that exacerbate inflammation. Increased oxidative stress also increases the production of inflammatory mediators [ 32 , 33 ]. Okkay U et al. found that IL-1Beta increased and IL-10 decreased with TAA application in their study [ 27 ]. In our study, IL-4, IL-6, IL-10, IL-1Beta and IL-1RA increased. IL-6, IL-10 and IL-1Beta increased less in the leukopenia-treated groups compared to the TAA-only group. This showed that lycopene is effective in the treatment of HE. 4.1. Limitation In our study, initial and final tests of locomotor activity were performed and compared to investigate the neurological efficacy of lycopene against HE, and blood ammonium levels and organ weights of the brain, liver and lungs were studied to assess organ protection. The results of this study reflect the promising effects of lycopene on HE. However, immunohistochemical analysis and molecular changes at the cellular level were not performed to determine whether the protective effect seen in our study was at the hepatic level, the neurological level, or both. In addition, since our study is an animal experimental study, human studies are needed to determine whether lycopene has a protective effect clinically. Because of these limitations, further studies are needed. 5. CONCLUSION To investigate the neurological efficacy of lycopene against HE, the locomotor activity tests and as a marker of neurological impairment blood ammonium levels and organ weights of the brain, liver and lungs were studied. Our locomotor activity test results in TAA-induced HE were associated with hyperammonemia, oxidative stress and inflammatory reactions in accordance with previous studies. Within the scope of our study, for the first time in the literature, whether lycopene administration is protective against the development of HE was investigated by animal experiment. According to the data obtained, it was determined that lycopene administration had hepatoprotective and neuroprotective efficacy against HE both in locomotor activity tests and at the liver level. Declarations Conflict of Interest: No conflict of interest to declare. Funding: None. Author Contribution Erdal Tekin: Conceptualization, original draft, methodology, formal analysis, writing, review, editing, software. Ersan Gursoy: Conceptualization, original draft, methodology, formal analysis, writing, review, editing. Fatma Tortum: Original draft, methodology, formal analysis, writing, review, editing. Mehmet Nuri Koçak: Methodology, formal analysis, writing, review, editing. Ufuk Okkay: Methodology, formal analysis, writing, review. Cemil Bayram: Methodology, review and editing. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6819328","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":471451628,"identity":"3b0b3526-abce-41dc-b852-83c07aff637f","order_by":0,"name":"Erdal Tekin","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABCUlEQVRIiWNgGAWjYHACA4YHQFKCmYHxAZjPDMQ8hLQkADFQCzOIBdciQVgLAwObBFgLAwEt/NKHN35IYPgjJ9nOe6zi5w+7xO3sQBe+bWOoM2/ArkWyL61YAmiLsTQzX9rNnoTkxJ3NDMyGc9sYJGQO4HDVGR4DkJbEecw8Zjd4EpgTNxxmYJPmBWrB5TL7MzzGP4Ba6kFaCv8k1IO0sP/Gp8WAh8cMZEuCNFALM0/CYbAtzPi0SJxhK7NIMDA2nNnMYywtk3bceMNhxmbJOeckJGfgCrEe5s03PlTIyUucP2P48Y1NteyG84cPfnhTZsOPL2JAUYMMGBsY8MfkKBgFo2AUjAJCAADKAUqL2yvK8wAAAABJRU5ErkJggg==","orcid":"","institution":"Atatürk University","correspondingAuthor":true,"prefix":"","firstName":"Erdal","middleName":"","lastName":"Tekin","suffix":""},{"id":471451629,"identity":"547b6f30-4932-4433-838c-8163f46892fe","order_by":1,"name":"Ersan Gursoy","email":"","orcid":"","institution":"Erzincan Binali Yıldırım University","correspondingAuthor":false,"prefix":"","firstName":"Ersan","middleName":"","lastName":"Gursoy","suffix":""},{"id":471451631,"identity":"a53fec2c-9382-46ca-bb7c-53f2317cf6c7","order_by":2,"name":"Fatma Tortum","email":"","orcid":"","institution":"Atatürk University","correspondingAuthor":false,"prefix":"","firstName":"Fatma","middleName":"","lastName":"Tortum","suffix":""},{"id":471451632,"identity":"e3d50dea-0527-4904-bad8-c52d7d3958f8","order_by":3,"name":"Mehmet Nuri Koçak","email":"","orcid":"","institution":"Atatürk University","correspondingAuthor":false,"prefix":"","firstName":"Mehmet","middleName":"Nuri","lastName":"Koçak","suffix":""},{"id":471451633,"identity":"5dcb68a8-df84-44fd-88a0-a3479ae66ce3","order_by":4,"name":"Ufuk Okkay","email":"","orcid":"","institution":"Atatürk University","correspondingAuthor":false,"prefix":"","firstName":"Ufuk","middleName":"","lastName":"Okkay","suffix":""},{"id":471451634,"identity":"7fd6cc11-35b3-4ec6-ada6-0891262803c0","order_by":5,"name":"Cemil Bayram","email":"","orcid":"","institution":"Atatürk University","correspondingAuthor":false,"prefix":"","firstName":"Cemil","middleName":"","lastName":"Bayram","suffix":""},{"id":471451635,"identity":"1d369ea9-2d40-47c6-8915-3e7eb8fb46e2","order_by":6,"name":"Zekai Halici","email":"","orcid":"","institution":"Atatürk University","correspondingAuthor":false,"prefix":"","firstName":"Zekai","middleName":"","lastName":"Halici","suffix":""},{"id":471451636,"identity":"31d17ead-e2bf-4a3d-b2aa-67805a7abcd8","order_by":7,"name":"Muhammet Celik","email":"","orcid":"","institution":"Atatürk University","correspondingAuthor":false,"prefix":"","firstName":"Muhammet","middleName":"","lastName":"Celik","suffix":""},{"id":471451637,"identity":"f6bb2045-d419-4919-8466-338a34c4520d","order_by":8,"name":"Ahmet Kızıltunç","email":"","orcid":"","institution":"Atatürk University","correspondingAuthor":false,"prefix":"","firstName":"Ahmet","middleName":"","lastName":"Kızıltunç","suffix":""}],"badges":[],"createdAt":"2025-06-04 10:23:22","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6819328/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6819328/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":84708816,"identity":"97634d41-06d9-4abb-8996-e6716a16dabd","added_by":"auto","created_at":"2025-06-16 12:55:17","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":214625,"visible":true,"origin":"","legend":"\u003cp\u003eExperimental procedure flow of the study.\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-6819328/v1/fb2aa000da0c306cefd8406a.png"},{"id":84708797,"identity":"5ff8a726-2735-4e5d-998a-8813fba341b2","added_by":"auto","created_at":"2025-06-16 12:55:17","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":21959,"visible":true,"origin":"","legend":"\u003cp\u003eComparison of weights (2a) and tissue weights (2b) of rats included in the study according to groups (Note: Weighing weights are given as mean±SD (n=6). Kruskal-Wallis test was used to compare the groups (p\u0026gt;0.05)).\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-6819328/v1/ae4e0c4fb02a54f46824046d.png"},{"id":84709195,"identity":"318319a0-2d08-40e6-ae49-506e10b67c72","added_by":"auto","created_at":"2025-06-16 13:03:17","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":36732,"visible":true,"origin":"","legend":"\u003cp\u003eBasal locomotor activity test results of rats (Note: Data are expressed as mean ± SD. a; p\u0026gt;0.05, b; p\u0026lt;0.05 vs. control).\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-6819328/v1/12be1c5c21fe296cab9640d9.png"},{"id":84709194,"identity":"844d35ac-f73f-41e3-b918-949afe4bd86b","added_by":"auto","created_at":"2025-06-16 13:03:17","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":34011,"visible":true,"origin":"","legend":"\u003cp\u003eFinal locomotor activity test results of rats (Note: Data are expressed as mean ± SD. a; p\u0026gt;0.05, c; p\u0026lt;0.05 vs. healthy).\u003c/p\u003e","description":"","filename":"floatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-6819328/v1/d554c7b03f966608389f82fe.png"},{"id":84709193,"identity":"25c8b087-4431-410b-b4e3-3e1d61b982e3","added_by":"auto","created_at":"2025-06-16 13:03:17","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":27564,"visible":true,"origin":"","legend":"\u003cp\u003eInterleukin analysis results of the groups\u003c/p\u003e","description":"","filename":"floatimage5.png","url":"https://assets-eu.researchsquare.com/files/rs-6819328/v1/31c56df88adb0b81f04c782f.png"},{"id":84708825,"identity":"6672b575-f88d-410c-99d0-e5c7b27422bc","added_by":"auto","created_at":"2025-06-16 12:55:17","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":22002,"visible":true,"origin":"","legend":"\u003cp\u003eBiochemical analysis results of the groups\u003c/p\u003e","description":"","filename":"floatimage6.png","url":"https://assets-eu.researchsquare.com/files/rs-6819328/v1/721437f2286e9bcb94f1b3b9.png"},{"id":86531101,"identity":"9bd2b575-13ce-4fa1-84bc-ec089f1a23a2","added_by":"auto","created_at":"2025-07-11 17:01:43","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":944164,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6819328/v1/c04f82d4-88ca-4269-8a64-d2d1996ec574.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Neuroprotective Efficacy of Lycopene in a Thioacetamide-Induced Model of Hepatic Encephalopathy: An Experimental Rat Study","fulltext":[{"header":"1. INTRODUCTION","content":"\u003cp\u003eLycopene is a powerful antioxidant that belongs to the carotenoid family and is found in red fruits and vegetables. Lycopene can be used in the treatment of many diseases and studies have shown that it has antioxidant, anti-inflammatory, anti-proliferative, anti-cancer and other properties [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Lycopene is hepatoprotective against several liver diseases, including alcoholic liver disease, nonalcoholic fatty liver disease, hepatic fibrosis, ischemia/reperfusion injury, hepatocellular carcinoma, fulminant liver failure and radiation injury [\u003cspan additionalcitationids=\"CR4\" citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Lycopene is thought to be a potent neuroprotectant against oxidative stress and could be used to prevent the injury or death of nerve cells in a number of neurological diseases [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Animal studies have highlighted its ability to adequately cross the blood-brain barrier and to improve many neurodegenerative disorders [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e], and another study found that lycopene was even neuroprotective against hypoxia-ischemic injury in rats [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eHepatic encephalopathy (HE) is a common complication of acute and chronic liver disease and is accompanied by neuropsychiatric disturbances. The pathogenesis of HE is multifactorial and has not yet been fully elucidated [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. However, morphological changes in astrocytes are thought to be caused by neurotoxic substances such as ammonium and manganese. This leads to brain dysfunction, brain atrophy and brain edema as a result of disruption of the blood-brain barrier [\u003cspan additionalcitationids=\"CR11\" citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. These metabolic abnormalities of HE are considered reversible. However, most patients are diagnosed after reversible brain damage has occurred, at which time neuropsychiatric disorders develop [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eAs HE has a high morbidity and mortality, new aspects need to be addressed to improve both liver and neurological disease. Lycopene may have improved HE because of its known antioxidant, anti-inflammatory, anti-proliferative and neuroprotective properties. This study aimed to evaluate the potential protective role of lycopene against HE and behavioral consequences in a rat model of thioacetamide-induced HE.\u003c/p\u003e"},{"header":"2. MATERIAL AND METHOD","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1. Study Design\u003c/h2\u003e \u003cp\u003e This study is an experimental study and the ethics committee approval for animal experiments was obtained from the Local Ethics Committee for Animal Experiments of our university (Decision No: 28.01.2021- 2021/1\u0026ndash;25). The handling of rats and experimental procedures were performed according to the guidelines for experimental animals.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2. Animals\u003c/h2\u003e \u003cp\u003eA total of 24 healthy albino male Sprague-Dawley rats weighing 250\u0026thinsp;\u0026plusmn;\u0026thinsp;25 g were used in the study. The rats were obtained from the Medical Experimental Application and Research Center of our university. The animals were housed in groups under controlled lighting conditions (12:12 h light/dark cycle) in a well-ventilated room at 22\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C in typical plastic cages on sawdust beds. Rats were fed ad libitum throughout the study.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3. Materials\u003c/h2\u003e \u003cp\u003eAn experimental HE model was established by intraperitoneal administration of thioacetamide (TAA) to rats. TAA is a sulfur-containing catalyst that causes liver fibrosis and hepatotoxicity by inducing activation of macrophages and secretion of various inflammatory cytokines [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Studies in the literature have found TAA to be a good agent in an experimentally induced HE model in rats. [\u003cspan additionalcitationids=\"CR15\" citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. TAA (Sigma-Aldrich, St. Louis, MO, USA) was prepared fresh by dissolving in sterile distilled water. Lycopene 10% FS (Redivivo, DSM Nutritional Products) was prepared by dissolution in corn oil and administered to the rats by gavage.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4. Study Groups\u003c/h2\u003e \u003cp\u003eA total of 24 rats were randomized into four groups of six rats each: healthy group, control group, low dose lycopene group and high dose lycopene group.\u003c/p\u003e \u003cp\u003eGroup 1: Healthy group (n\u0026thinsp;=\u0026thinsp;6); no treatment was administered and only weight was monitored.\u003c/p\u003e \u003cp\u003eGroup 2: Control group (n\u0026thinsp;=\u0026thinsp;6); weight was monitored for 15 days and 200 mg/kg TAA was administered intraperitoneally for 3 consecutive days from day 15 to day 17. On the days of TAA administration, fluid replacement was performed with lactated ringer (10 mg/kg/day) in the morning and evening.\u003c/p\u003e \u003cp\u003eGroup 3: Low-dose lycopene group (50 mg/kg, n\u0026thinsp;=\u0026thinsp;6); weight was monitored for 15 days and lycopene 50 mg/kg was administered by gavage at the same time every day until the 15th day of the experiment. From the 15th to the 17th day of the experiment, 200 mg/kg TAA was administered intraperitoneally every day for 3 consecutive days. On the days of TAA administration, fluid replacement with lactated ringer (10 mg/kg/day) was performed in the morning and evening.\u003c/p\u003e \u003cp\u003eGroup 4: High-dose lycopene group (100 mg/kg, n\u0026thinsp;=\u0026thinsp;6); body weights were monitored for 15 days and lycopene 100 mg/kg was administered by gavage at the same time every day until the 15th day. From the 15th to the 17th day of the experiment, 200 mg/kg TAA was administered intraperitoneally every day for 3 consecutive days. On the days of TAA administration, fluid replacement was performed in the morning and evening with lactated ringer (10 mg/kg/day).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.5. Locomotor Activity Test\u003c/h2\u003e \u003cp\u003eThe locomotor activity test was measured with an open-field activity imager system (May Act 508) on the first day of the study to determine baseline measurements and again on day 18 at the end of the study to determine changes.\u003c/p\u003e \u003cp\u003eThe basis of the system used to measure locomotor activity is a square quadrilateral with infrared light sources on each side. A square Plexiglas cage is placed inside this quadrilateral. When the animal makes a movement inside the cage, it interrupts the communication between the opposite infrared sensors and this is recorded by a recorder connected to the device according to the shape of the movement made by the subject. With the help of this system, horizontal, vertical and ambulatory activities of rats are recorded. Horizontal movement is the movement of the experimental animal in place without any displacement or standing movement. Vertical movement is a standing movement and is detected by vertical sensors on the bands. Ambulatory movement is any displacement movement of the animal in the cage other than standing. Horizontal and vertical activities give an idea of the animal's stereotypical movements and aggressiveness. All three activities can be evaluated separately or the sum of the three can be expressed as total locomotor activity. Resting times and distance traveled were measured and recorded. All rats were placed individually in a transparent cage twice, once at the beginning and once at the end of the study, and recorded for 10 minutes to determine whether locomotor activity decreased or increased. The flow chart of the study is shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e2.6. Study Protocol\u003c/h2\u003e \u003cp\u003eThe same preoperative preparation, anesthesia and surgical technique were used for each group. All rats were euthanized under ketamine/xylazine anesthesia when the study was completed. Blood samples were collected for biochemical and hematological analysis. Liver, brain and lungs of all rats were removed, weighed and compared to investigate possible infection and inflammation in the organs.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e2.7. Biochemical analysis\u003c/h2\u003e \u003cp\u003eBlood samples (5 cc) were collected from all rats, clotted at room temperature, then placed in a yellow-capped gel tube and centrifuged at 3000 rpm for 10 minutes. Serum samples were placed in Eppendorf tubes and analyzed for biochemical parameters. Additionally, blood samples of 3cc were collected from all rats and placed in EDTA-containing hemogram tubes according to the cold chain rules, and ammonium analysis was performed immediately after plasma separation.\u003c/p\u003e \u003cp\u003eAU 5800 Beckman coulter autoanalyzer\u0026reg; (USA) and standard kits were used to measure serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin, direct bilirubin and plasma ammonium levels. Bioassay Technology Laboratory (BT LAB\u0026reg;, Shanghai, Korain) rat kits were used for ELISA (enzyme-linked immunosorbent assay) analysis of interleukin (IL) 1beta, IL-1RA, IL-4, IL-6 and IL-10 levels.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e2.8. Statistical Analysis\u003c/h2\u003e \u003cp\u003eIn our study, statistical analyses were performed with SPSS 25.0 (IBM Corp., NY, USA). Categorical data were presented as percentage and frequency and numerical data as mean and standard deviation. Kolmogorov-Smirnov test was used for normal distribution evaluation. Mann Whitney U test and Kruskal Wallis variance test were used for statistical analysis between groups. Dunnett T3 was used for post-hoc analysis of the data. Statistical significance was taken as p\u0026thinsp;\u0026lt;\u0026thinsp;0.05 in the whole study.\u003c/p\u003e \u003c/div\u003e"},{"header":"3. RESULTS","content":"\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e3.1. Weights of the rat and its organs\u003c/h2\u003e \u003cp\u003eThe mean weights of all groups that were weighed every three days within the scope of our study are shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e2\u003c/span\u003ea. The mean weights of the rats included in the study were 277.67\u0026thinsp;\u0026plusmn;\u0026thinsp;9.32 and there was no statistically significant difference (p\u0026thinsp;=\u0026thinsp;0.172). After euthanasia, the organs of the rats were removed and the weighed organ weights are shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e2\u003c/span\u003eb. Accordingly, there was no significant difference in organ weights between groups (lung, liver and brain p values were 0.254, 0.371, 0.903, respectively).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003e3.2. Locomotor activity test results\u003c/h2\u003e \u003cp\u003eLocomotor activity tests were performed in all rats included in the study both at the beginning and at the end of the study. Basal locomotor tests are given in Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e3\u003c/span\u003e. Accordingly, when the control group was compared with the other groups, stereotypic movement and resting time duration were higher in the control group, while other locomotor activity movements were lower. While there was no statistical significance between the groups in stereotypic, ambulatory and vertical locomotor activity tests (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05), there was statistical significance in other locomotor activities (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e \u003cp\u003eThe final locomotor tests of the rats are given in Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e4\u003c/span\u003e. According to this, while there was no statistical significance in stereotypic movements (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05), other locomotor activity tests were statistically significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Ambulatory, vertical, and horizontal movement were higher in healthy rats and the healthy group was statistically significant compared to the other groups (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Resting time was less in healthy rats and statistically significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003e3.3. Biochemical analysis results\u003c/h2\u003e \u003cp\u003eThe results of interleukin analysis in the blood samples of the rats taken at the end of the study are given in Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e5\u003c/span\u003e. Accordingly, there was no statistical significance in IL-4 and IL-1RA analysis results (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05). IL-6, IL-10 and IL-1ꞵ analysis results were significant and as a result of the post-hoc analysis, there was statistical significance between the control group and the healthy and high dose lycopene group (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e \u003cp\u003eBiochemical analysis results of the blood samples of the rats taken at the end of the study are given in Fig.\u0026nbsp;\u003cspan refid=\"Fig9\" class=\"InternalRef\"\u003e6\u003c/span\u003e. Accordingly, there was statistical significance in the analysis results (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). It was determined that AST, ALT, total bilirubin, direct bilirubin and ammonia parameters were lower in the low dose lycopene group compared to the control and high dose lycopene group, which was statistically significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e \u003c/div\u003e"},{"header":"4. DISCUSSION","content":"\u003cp\u003eIn this study, weight and organ weights of HE-induced rats were calculated and locomotor activity tests and biochemical analyses were performed for behavioral defects. As a result of this study, it was determined that the weight of weight increased in the low dose lycopene group, while liver weight increased in the control group. Locomotor activity tests generally decreased but total distance traveled increased in both groups treated with lycopene. In addition, it was determined that IL-6, IL10, IL-1Beta, ALT, AST and ammonia levels decreased in both groups treated with lycopene compared to the control group. These results showed that lycopene has a protective effect against THA-induced HE.\u003c/p\u003e \u003cp\u003eIn HE, excessive amounts of ammonia cross the blood-brain barrier and disrupt brain metabolism and astrocyte structure, leading to cognitive and motor symptoms. With hyperammonemia, neuroinflammation occurs, contributing to intellectual disability [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. Lycopene has the ability to counteract neuroinflammation, proteinopathies, apoptosis, synaptic dysfunction and brain edema, as well as ameliorating oxidative stress. In the literature, these neuroprotective properties of lycopene have been emphasized to ameliorate cognitive deficits, locomotor disorders, dementia, seizures, anxiety and behavioral abnormalities including depression-like behaviors associated with neurological disorders [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. In our study, there was deterioration in the locomotor activity tests of the other groups compared to the healthy group in animal models of HE. Locomotor activity test results in both groups treated with lycopene were better than the control group. This shows that lycopene improves locomotor activity tests in HE. In addition, in our study, ammonia levels were found to be lower in the lycopene -treated groups compared to the untreated group. This shows us that leukopenia can treat high ammonia levels in HE. Thus, we think that lycopene can be used in the treatment of cognitive and cognitive functions caused by high ammonia levels.\u003c/p\u003e \u003cp\u003eHE occurs as a result of the interaction of many pathophysiologic factors in brain metabolism such as damage in blood-brain barrier permeability, inflammation and oxidative stress [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e, \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. Most of the drugs used to treat HE also aim to reduce blood serum ammonia and suppress neurotoxin production [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. In addition, inflammation and oxidative stress accelerate the progression of HE [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. Antioxidant and anti-inflammatory drugs have been reported to have protective effects on the brain and liver [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. There are no studies investigating the efficacy of lycopene, which has antioxidant and anti-inflammatory properties, in HE. In this study, we aimed to investigate this efficacy and this is the first study in the literature. As the results of our study showed that locomotor activity test results improved, it was concluded that lycopene improves cognitive function damage in HE. In the literature, it has been emphasized that lycopene has hepatoprotective properties against liver diseases and improves memory and cognition abilities of rodents in animal experiments [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. In the HE model; it was determined that lycopene administered orally for 15 days had hepatoprotective and cognitive enhancing effect in THA-induced HE and this is the first study showing this effect.\u003c/p\u003e \u003cp\u003eIn the literature, there are studies reporting that liver enzyme levels of rats are impaired in THA-induced HE. In these studies, it was emphasized that ALT and AST levels increased and ammonia concentration increased. These studies show the severity of liver damage [\u003cspan additionalcitationids=\"CR28 CR29 CR30\" citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. Similar to the studies in the literature, hepatic enzyme activity and ammonia level increased in our study. In addition, liver damage was attenuated with leukopenia treatment.\u003c/p\u003e \u003cp\u003eCognitive and neurological impairments detected in HE models were associated with increased ammonia levels, oxidative stress and pro-inflammatory cytokines that exacerbate inflammation. Increased oxidative stress also increases the production of inflammatory mediators [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. Okkay U et al. found that IL-1Beta increased and IL-10 decreased with TAA application in their study [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. In our study, IL-4, IL-6, IL-10, IL-1Beta and IL-1RA increased. IL-6, IL-10 and IL-1Beta increased less in the leukopenia-treated groups compared to the TAA-only group. This showed that lycopene is effective in the treatment of HE.\u003c/p\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003e4.1. Limitation\u003c/h2\u003e \u003cp\u003eIn our study, initial and final tests of locomotor activity were performed and compared to investigate the neurological efficacy of lycopene against HE, and blood ammonium levels and organ weights of the brain, liver and lungs were studied to assess organ protection. The results of this study reflect the promising effects of lycopene on HE. However, immunohistochemical analysis and molecular changes at the cellular level were not performed to determine whether the protective effect seen in our study was at the hepatic level, the neurological level, or both. In addition, since our study is an animal experimental study, human studies are needed to determine whether lycopene has a protective effect clinically. Because of these limitations, further studies are needed.\u003c/p\u003e \u003c/div\u003e"},{"header":"5. CONCLUSION","content":"\u003cp\u003eTo investigate the neurological efficacy of lycopene against HE, the locomotor activity tests and as a marker of neurological impairment blood ammonium levels and organ weights of the brain, liver and lungs were studied. Our locomotor activity test results in TAA-induced HE were associated with hyperammonemia, oxidative stress and inflammatory reactions in accordance with previous studies. Within the scope of our study, for the first time in the literature, whether lycopene administration is protective against the development of HE was investigated by animal experiment. According to the data obtained, it was determined that lycopene administration had hepatoprotective and neuroprotective efficacy against HE both in locomotor activity tests and at the liver level.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eConflict of Interest:\u003c/h2\u003e\n\u003cp\u003eNo conflict of interest to declare.\u003c/p\u003e\n\u003ch2\u003eFunding:\u003c/h2\u003e\n\u003cp\u003eNone.\u003c/p\u003e\n\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\n\u003cp\u003eErdal Tekin: Conceptualization, original draft, methodology, formal analysis, writing, review, editing, software. Ersan Gursoy: Conceptualization, original draft, methodology, formal analysis, writing, review, editing. Fatma Tortum: Original draft, methodology, formal analysis, writing, review, editing. Mehmet Nuri Ko\u0026ccedil;ak: Methodology, formal analysis, writing, review, editing. Ufuk Okkay: Methodology, formal analysis, writing, review. Cemil Bayram: Methodology, review and editing. Zekai Halici: Methodology, formal analysis, writing, review, editing and original draft. Muhammet \u0026Ccedil;elik: Writing, original draft, software and editing. Ahmet Kızıltun\u0026ccedil;: original draft, software and editing.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eStice CP, Xia H, Wang XD: Tomato lycopene prevention of alcoholic fatty liver disease and hepatocellular carcinoma development. \u003cem\u003eChronic Dis Transl Med \u003c/em\u003e2018, 4(4):211-224.\u003c/li\u003e\n\u003cli\u003eIbrahim IM, Althagafy HS, Abd-Alhameed EK, Al-Thubiani WS, Hassanein EHM: Promising hepatoprotective effects of lycopene in different liver diseases. \u003cem\u003eLife Sci \u003c/em\u003e2022, 310:121131.\u003c/li\u003e\n\u003cli\u003eRao AV, Rao LG: Carotenoids and human health. \u003cem\u003ePharmacological Research \u003c/em\u003e2007, 55(3):207-216.\u003c/li\u003e\n\u003cli\u003eChen D, Huang C, Chen Z: A review for the pharmacological effect of lycopene in central nervous system disorders. \u003cem\u003eBiomed Pharmacother \u003c/em\u003e2019, 111:791-801.\u003c/li\u003e\n\u003cli\u003eIcel E, Icel A, U\u0026ccedil;ak T, Karakurt Y, Elpeze B, Keskin \u0026Ccedil;imen F, S\u0026uuml;leyman H: The effects of lycopene on alloxan induced diabetic optic neuropathy. \u003cem\u003eCutaneous and Ocular Toxicology \u003c/em\u003e2019, 38(1):88-92.\u003c/li\u003e\n\u003cli\u003eOguz E, Terzioglu Bebitoglu B, Acet G, Hodzic A, Hatiboglu N, Ada S: Effect of lycopene on As(2)O(3) induced oxidative stress in SH-SY5Y cells. \u003cem\u003eMol Biol Rep \u003c/em\u003e2021, 48(4):3205-3212.\u003c/li\u003e\n\u003cli\u003ePaul R, Mazumder MK, Nath J, Deb S, Paul S, Bhattacharya P, Borah A: Lycopene - A pleiotropic neuroprotective nutraceutical: Deciphering its therapeutic potentials in broad spectrum neurological disorders. \u003cem\u003eNeurochem Int \u003c/em\u003e2020, 140:104823.\u003c/li\u003e\n\u003cli\u003eFu C, Zheng Y, Zhu J, Chen B, Lin W, Lin K, Zhu J, Chen S, Li P, Fu X\u003cem\u003e et al\u003c/em\u003e: Lycopene Exerts Neuroprotective Effects After Hypoxic-Ischemic Brain Injury in Neonatal Rats via the Nuclear Factor Erythroid-2 Related Factor 2/Nuclear Factor-\u0026kappa;-Gene Binding Pathway. \u003cem\u003eFront Pharmacol \u003c/em\u003e2020, 11:585898.\u003c/li\u003e\n\u003cli\u003eOchoa-Sanchez R, Tamnanloo F, Rose CF: Hepatic Encephalopathy: From Metabolic to Neurodegenerative. \u003cem\u003eNeurochemical Research \u003c/em\u003e2021, 46(10):2612-2625.\u003c/li\u003e\n\u003cli\u003eRudler M, Weiss N, Bouzbib C, Thabut D: Diagnosis and Management of Hepatic Encephalopathy. \u003cem\u003eClin Liver Dis \u003c/em\u003e2021, 25(2):393-417.\u003c/li\u003e\n\u003cli\u003eWeiss N, Jalan R, Thabut D: Understanding hepatic encephalopathy. \u003cem\u003eIntensive Care Med \u003c/em\u003e2018, 44(2):231-234.\u003c/li\u003e\n\u003cli\u003eBraissant O, McLin VA, Cudalbu C: Ammonia toxicity to the brain. \u003cem\u003eJournal of Inherited Metabolic Disease \u003c/em\u003e2013, 36(4):595-612.\u003c/li\u003e\n\u003cli\u003eRavichandra A, Schwabe RF: Mouse Models of LiverliversFibrosis. 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Toxicol \u003c/em\u003e2021, 35(5):e22735.\u003c/li\u003e\n\u003cli\u003eOng JP, Aggarwal A, Krieger D, Easley KA, Karafa MT, Van Lente F, Arroliga AC, Mullen KD: Correlation between ammonia levels and the severity of hepatic encephalopathy. \u003cem\u003eAm J Med \u003c/em\u003e2003, 114(3):188-193.\u003c/li\u003e\n\u003cli\u003eCrowe-White KM, Phillips TA, Ellis AC: Lycopene and cognitive function. \u003cem\u003eJournal of nutritional science \u003c/em\u003e2019, 8:e20.\u003c/li\u003e\n\u003cli\u003eZhu N-W, Yin X-L, Lin R, Fan X-L, Chen S-J, Zhu Y-M, Zhao X-Z: Possible mechanisms of lycopene amelioration of learning and memory impairment in rats with vascular dementia. \u003cem\u003eNeural Regeneration Research \u003c/em\u003e2020, 15(2):332.\u003c/li\u003e\n\u003cli\u003eHadjihambi A, Arias N, Sheikh M, Jalan R: Hepatic encephalopathy: a critical current review. \u003cem\u003eHepatology international \u003c/em\u003e2018, 12:135-147.\u003c/li\u003e\n\u003cli\u003eOja SS, Saransaari P, Korpi ER: Neurotoxicity of ammonia. \u003cem\u003eNeurochemical research \u003c/em\u003e2017, 42:713-720.\u003c/li\u003e\n\u003cli\u003eMoran S, L\u0026oacute;pez-S\u0026aacute;nchez M, Milke-Garc\u0026iacute;a MDP, Rodr\u0026iacute;guez-Leal G: Current approach to treatment of minimal hepatic encephalopathy in patients with liver cirrhosis. \u003cem\u003eWorld J Gastroenterol \u003c/em\u003e2021, 27(22):3050-3063.\u003c/li\u003e\n\u003cli\u003eManzhalii E, Virchenko O, Falalyeyeva T, Moiseienko V, Nykula T, Kondratiuk V, Savchuk O, Beregova T, Stremmel W: Hepatic encephalopathy aggravated by systemic inflammation. \u003cem\u003eDigestive Diseases \u003c/em\u003e2019, 37(6):509-517.\u003c/li\u003e\n\u003cli\u003eG\u0026uuml;ven M, Aras AB, Topaloğlu N, \u0026Ouml;zkan A, Şen HM, Kalkan Y, Okuyucu A, Akbal A, G\u0026ouml;kmen F, Coşar M: The protective effect of syringic acid on ischemia injury in rat brain. \u003cem\u003eTurkish journal of medical sciences \u003c/em\u003e2015, 45(1):233-240.\u003c/li\u003e\n\u003cli\u003eSingh S, Murad MH, Chandar AK, Bongiorno CM, Singal AK, Atkinson SR, Thursz MR, Loomba R, Shah VH: Comparative Effectiveness of Pharmacological Interventions for Severe Alcoholic Hepatitis: A Systematic Review and Network Meta-analysis. \u003cem\u003eGastroenterology \u003c/em\u003e2015, 149(4):958-970.e912.\u003c/li\u003e\n\u003cli\u003eOkkay U, Ferah Okkay I, Cicek B, Aydin IC, Ozkaraca M: Hepatoprotective and neuroprotective effect of taxifolin on hepatic encephalopathy in rats. \u003cem\u003eMetabolic Brain Disease \u003c/em\u003e2022, 37(5):1541-1556.\u003c/li\u003e\n\u003cli\u003eYogalakshmi B, Viswanathan P, Anuradha CV: Investigation of antioxidant, anti-inflammatory and DNA-protective properties of eugenol in thioacetamide-induced liver injury in rats. \u003cem\u003eToxicology \u003c/em\u003e2010, 268(3):204-212.\u003c/li\u003e\n\u003cli\u003eFerah Okkay I, Okkay U, Gundogdu OL, Bayram C, Mendil AS, Ertugrul MS, Hacimuftuoglu A: Syringic acid protects against thioacetamide-induced hepatic encephalopathy: Behavioral, biochemical, and molecular evidence. \u003cem\u003eNeurosci Lett \u003c/em\u003e2022, 769:136385.\u003c/li\u003e\n\u003cli\u003eKhodir AE, Said E: Nifuroxazide attenuates experimentally-induced hepatic encephalopathy and the associated hyperammonemia and cJNK/caspase-8/TRAIL activation in rats. \u003cem\u003eLife Sci \u003c/em\u003e2020, 252:117610.\u003c/li\u003e\n\u003cli\u003eBayramoglu G, Bayramoglu A, Altuner Y, Uyanoglu M, Colak S: The effects of lycopene on hepatic ischemia/reperfusion injury in rats. \u003cem\u003eCytotechnology \u003c/em\u003e2015, 67(3):487-491.\u003c/li\u003e\n\u003cli\u003eLu L, Wu C, Lu B-j, Xie D, Wang Z, Azami NLB, An Y-t, Wang H-j, Ye G, Sun M-y: BabaoDan cures hepatic encephalopathy by decreasing ammonia levels and alleviating inflammation in rats. \u003cem\u003eJournal of ethnopharmacology \u003c/em\u003e2020, 249:112301.\u003c/li\u003e\n\u003cli\u003eHernandez-Rabaza V, Cabrera-Pastor A, Taoro-Gonzalez L, Gonzalez-Usano A, Agusti A, Balzano T, Llansola M, Felipo V: Neuroinflammation increases GABAergic tone and impairs cognitive and motor function in hyperammonemia by increasing GAT-3 membrane expression. Reversal by sulforaphane by promoting M2 polarization of microglia. \u003cem\u003eJ Neuroinflammation \u003c/em\u003e2016, 13(1):83.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Experimental study, hepatic encephalopathy, lycopene, neuroprotection","lastPublishedDoi":"10.21203/rs.3.rs-6819328/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6819328/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eObjective\u003c/h2\u003e \u003cp\u003eLycopene has known antioxidant, anti-inflammatory, anti-proliferative, and neuroprotective effects. This is the first study to evaluate the potential protective role of lycopene against hepatic encephalopathy.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e \u003cp\u003eAn experimental hepatic encephalopathy model was established by intraperitoneal administration of thioacetamide to rats. A total of 24 rats were randomized into four groups: healthy group, control group, low-dose (50 mg/kg) lycopene group, and high-dose (100 mg/kg) lycopene group. The locomotor activity test was measured on the first day of the study to determine baseline measurements and again at the end of the study to determine changes. Blood samples were collected, and liver, brain, and lungs were removed and weighed.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eExcept for stereotypic movements (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05), all other final locomotor activity tests were statistically significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). There was no statistical significance in IL-4 and IL-1RA analysis results (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05), but IL-6, IL-10 and IL-1ꞵ analysis results were significant between control and healthy and high-dose lycopene groups (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). AST, ALT, total bilirubin, direct bilirubin and ammonia parameters were significantly lower in the low-dose lycopene group compared to the control and high-dose lycopene groups (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eLycopene administration had hepatoprotective and neuroprotective efficacy against HE, supported both in locomotor activity tests and at biochemical tests.\u003c/p\u003e","manuscriptTitle":"Neuroprotective Efficacy of Lycopene in a Thioacetamide-Induced Model of Hepatic Encephalopathy: An Experimental Rat Study","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-06-16 12:55:12","doi":"10.21203/rs.3.rs-6819328/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"ba4fa5ce-9890-457f-9e3c-6e1b5909a502","owner":[],"postedDate":"June 16th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-07-11T16:53:36+00:00","versionOfRecord":[],"versionCreatedAt":"2025-06-16 12:55:12","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6819328","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6819328","identity":"rs-6819328","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}