Dose Of CCl4 Research Articles

Evaluation of the therapeutic potential of novel nanoparticle formulations of glutathione and virgin coconut oil in an experimental model of carbon tetrachloride-induced liver failure.

Acute liver failure (ALF) is a critical condition characterized by rapid liver dysfunction, leading to high mortality rates. Current treatments are limited, primarily supportive, and often require liver transplantation. This study investigates the potential of a novel nanoparticle formulation of glutathione (GSH) and virgin coconut oil (VCO) alone and in combination to enhance therapeutic outcomes in a rat model of ALF induced by orogastric carbon tetrachloride (CCl4). The study employed adult male Albino rats divided into ten groups, with ALF induced via a single oral dose of CCl4. Various treatment regimens were administered over seven days, including conventional and nanoparticle forms of GSH and VCO and their combinations. The efficacy of treatments was evaluated through biochemical analysis of liver function markers, oxidative stress indicators, inflammatory biomarkers, and histopathological examinations. Nanoparticles were synthesized using established methods, and characterization techniques were employed to ensure their quality and properties. The nanoparticle formulations significantly improved liver function, as indicated by reduced serum levels of alanine aminotransferase and aspartate aminotransferase, alongside decreased oxidative stress markers such as malondialdehyde. Furthermore, they reduced tumor necrosis factor alpha and interleukin-1 beta inflammatory markers. Histological analysis revealed reduced hepatocellular necrosis and inflammation in treated groups compared to controls. Also, decreased nuclear factor-kappa B was detected by immunohistochemical analysis. The findings show that the nanoparticle mixture of GSH and VCO effectively reduces liver damage in ALF. This suggests a promising drug-based approach for improving liver regeneration and protection. This innovative strategy may pave the way for new therapeutic interventions in the management of ALF.

BIOCHEMICAL AND HISTOLOGICAL STUDY OF THREE DIFFERENT MULTIHERBAL COMPOSITIONS ON HEPATOXICITY INDUCED BY CARBON TETRA CHLORIDE (CCL4) IN ALBINO RATS

Objective: The present study aimed to evaluate the hepatoprotective activity of three different types of multiherbal formulations in comparison to standard drug (Silymarin). Methods: Five hepatoprotective plants were chosen to make three different kinds of formulations, as mentioned in the materials and methods. These five selected herbs were chosen based on previous researches or reviews on hepatoprotective herbs. For the present study, two different groups of rats were made: the control group and the experimental group. The experimental group was further divided into five subgroups. Hepatotoxicity was induced by a single oral dose (1.5 ml/kg) of CCl4. After this, all rats were treated with different formulations and standard drug (silymarin) according to their groups. Samples for biochemical and histopathological (liver sample) examinations were collected on the fixed schedules (07th, 14th, and 21st d). Results: Biochemical parameters like SGPT, SGOT, and ALP were elevated due to CCl4-induced hepatotoxicity, and after treatment, they were recouped to normal significantly (P<0.001) by the treatment of formulation-I. On the other hand, decreased serum proteins like total, albumin, and globulin were increased to be normal (P<0.01) with the treatment of formulation-I. The histopathological study also supported the final results of the biochemical analysis. The damaged hepatocellular structures were repaired by the F-I formulation better than other formulations. Conclusion: The final results suggested that Formulation-I is a better healer than other formulations (F-II and F-III). It was thus concluded in the present study that formulation-I has better hepatoprotective activity than formulation-II and formulation-III.

Levisticum officinale extract protects against CCl4-induced hepatotoxicity through anti-inflammatory, anti-fibrotic, and antioxidant properties in rats

Objective: To investigate the hepatoprotective effects of Levisticum officinale extract on CCl4-induced hepatotoxicity. Methods: Different doses of Levisticum officinale extract were given orally to rats for 10 days, then rats received a single dose of CCl4 (2.5 mL/kg, 50% v/v in liquid paraffin). Biochemical and histopathological assays were performed to assess the effects of the extract on liver function and architecture. Moreover, antioxidant and oxidative markers as well as inflammatory and fibrotic indicators were measured. Results: Pretreatment with Levisticum officinale extract significantly mitigated CCl4-induced damage to liver structure, improved serum levels of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, urea, total bilirubin, and total protein, enhanced glutathione content and superoxide dismutase and catalase activities in the liver, as well as decreased plasma and hepatic malondialdehyde levels. Immunohistochemical results demonstrated that the extract reduced Ki-67 and α-SMA expression and Masson’s trichrome staining revealed decreased liver collagen in rats treated with Levisticum officinale extract. Moreover, Levisticum officinale extract markedly decreased the gene expressions of TNF-α, 1L-6, TGF-β1, MCP-1, and COX-2. Conclusions: Levisticum officinale extract exerts hepatoprotective effects on CCl4-induced hepatotoxicity through antioxidant, anti-inflammatory, and anti-fibrotic activities.

Abstract 7443: Investigating the impact of ferroptosis in a high-fat and high-oxidative stress hepatic steatosis environment

Abstract An iron-dependent mechanism of cell death, ferroptosis, occurs with intracellular iron accumulation and is associated with lipid peroxidation. Previous studies have shown that lipid peroxidation-induced ROS leads to DNA damage and promotes the progression of steatosis to advanced HCC. Therefore, we aim to study ferroptosis in a cell model with hepatocytes co-cultured with ACM (adipocyte culture medium) and low dose H2O2 for a high-fat/high ROS environment in vitro; an animal model with high-fat content and accompanying low dose carbon tetrachloride in vivo to observe the possibility of hepatocyte steatosis due to ferroptosis. Animal model: Mice were fed a Western diet and provided with high fructose/glucose-containing sugar drinking water (WFSD) for 7 months while receiving a monthly extra-low dose of CCl4. Tissue staining: Liver sections were performed to observe histological abnormalities in each group. Cell model: Pre-adipocytes, 3T3L1 were differentiated by adding IBMX, Dexamethasone, and Insulin (10 μg/ml). After differentiation, ACM was collected. The XTT assay was used to examine the relationship between cell death and ferroptosis by a ferroptosis inducer (Erastin) in high-fat and high-ROS environments.qPCR: To detect the expression of iron transport-related genes.Western Blot: To ascertain protein levels associated with ferroptosis. Mitochondrial assay: Mitochondrial membrane potential (MMP), mitochondrial dynamics-related proteins, and fluorescence mitochondrial tracking and staining. Ferritin, GPX4, IRP2, and ferroportin were upregulated when ACM co-culture with the hepatocytic AML12 cell line, and it also increased the sensitivity of ROS-related cell cytotoxicity, which was related to ferroptosis by the inducer, Erastin, in the high-fat/high-ROS cell environment. However, these results can be suppressed by UAMC3203, a ferroptosis inhibitor. As for the animal model with high-fat and carbon tetrachloride accompanied experiments, the mice had higher weight, Alanine Aminotransferase (ALT) levels, and developed tumors. Their liver sections showed lipid accumulation, histological fibrosis, and iron accumulation in both the WD and WD+CCl4 groups, which corresponds to the IRP1 and IRP2 upregulated in both groups by qPCR result. Our research findings demonstrate that ferroptosis correlates with high-fat and high-ROS-induced hepatocellular damage. The related mitochondria dynamics changes in liver tissue and high-fat and/or high ROS provide evidence for ferroptosis-related hepatic steatosis. We have established the effect of ferroptosis on hepatic steatosis in vivo and in vitro in nonalcoholic steatosis hepatitis (NASH). These results may enable the development of precise health prevention strategies by inhibiting ferroptosis to prevent the occurrence of liver diseases including hepatic steatosis and HCC. Citation Format: Chi-Sheng (Wayne) Chen, Shu-Chi Wang. Investigating the impact of ferroptosis in a high-fat and high-oxidative stress hepatic steatosis environment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7443.

Early detection of liver injuries by the Serum enhanced binding test sensitive to albumin post-transcriptional modifications

Early and sensitive biomarkers of liver dysfunction and drug-induced liver injury (DILI) are still needed, both for patient care and drug development. We developed the Serum Enhanced Binding (SEB) test to reveal post-transcriptional modifications (PTMs) of human serum albumin resulting from hepatocyte dysfunctions and further evaluated its performance in an animal model. The SEB test consists in spiking serum ex-vivo with ligands having specific binding sites related to the most relevant albumin PTMs and measuring their unbound fraction. To explore the hypothesis that albumin PTMs occur early during liver injury and can also be detected by the SEB test, we induced hepatotoxicity in male albino Wistar rats by administering high daily doses of ethanol and CCl4 over several days. Blood was collected for characterization and quantification of albumin isoforms by high-resolution mass spectrometry, for classical biochemical analyses as well as to apply the SEB test. In the exposed rats, the appearance of albumin isoforms paralleled the positivity of the SEB test ligands and histological injuries. These were observed as early as D3 in the Ethanol and CCl4 groups, whereas the classical liver tests (ALT, AST, PAL) significantly increased only at D7. The behavior of several ligands was supported by structural and molecular simulation analysis. The SEB test and albumin isoforms revealed hepatocyte damage early, before the current biochemical biomarkers. The SEB test should be easier to implement in the clinics than albumin isoform profiling.

Influence of omega- 3 fatty acids, soya isoflavones and their combination for abrogating carbon tetrachloride hazards in male rats.

Studies have shown that carbon tetrachloride (CCl4) induces hepatic and renal damage arising from oxidative stress. The present study was undertaken to examine the effect of omega-3 fatty acids and/or soya isoflavones on CCl4 induced toxicity in male albino rat liver and kidney. For this purpose, 42 rats were divided as follows: group 1, rats serves as the control without any treatment; group 2, rats were administered a single dose of CCl4 intraperitoneally (1 mg/kg b. wt.); group 3, rats were supplemented daily with omega-300 orally (400 mg/kg b. wt.); group 4, rats were supplemented daily with pro-S orally (50 mg/kg b. wt.); group 5, rats were supplemented daily with omega-300 orally for four weeks, then after 24 hours treated with a single dose of CCl4 at the same tested doses. group 6, rats were supplemented daily with pro- S orally for four weeks, then after 24 hours treated with a single dose of CCl4 at the same tested doses; group 7, rats were supplemented daily with an oral combination of omega-300 and pro-S orally for four weeks, then after 24 hours treated with a single dose of CCl4 at the same tested doses. Results showed that CCl4 administration induces hepatic damage indicated by a significant increase in the activities of alkaline phosphatase (ALP), aspartate aminotransferase (AST) and Aalanine aminotransferase (ALT) enzymes and glucose level, with a significant increase in malondialdehyde (MDA) and nitric oxide (NO) levels and a significant decrease of reduced glutathione (GSH) level in liver tissue. Also, CCl4 toxicity induce renal damage manifested in a significant increase in serum urea, creatinine, uric acid, and oxidative stress of kidney tissue reflected by increase of MDA, NO and the decrease of GSH levels. The pre-treatment with omega-3 fatty acids and/or soya isoflavones revealed ameliorative effect against deleterious effects of CCl4 toxicity on hepatic and renal tissues and all tested parameters. Results of the current study revealed also that the pre-treatment with omega-3 fatty acids and/or soya isoflavones to rats improved liver and kidney function and produced high antioxidant activity.

In Vivo and In Silico Studies of the Hepatoprotective Activity of Tert-Butylhydroquinone.

Tert-butylhydroquinone (TBHQ) is a synthetic food antioxidant with biological activities, but little is known about its pharmacological benefits in liver disease. Therefore, this work aimed to evaluate TBHQ during acute liver damage induced by CCl4 (24 h) or BDL (48 h) in Wistar rats. It was found that pretreatment with TBHQ prevents 50% of mortality induced by a lethal dose of CCl4 (4 g/kg, i.p.), and 80% of BDL+TBHQ rats survived, while only 50% of the BDL group survived. Serum markers of liver damage and macroscopic and microscopic (H&E staining) observations suggest that TBHQ protects from both hepatocellular necrosis caused by the sublethal dose of CCl4 (1.6 g/kg, i.p.), as well as necrosis/ductal proliferation caused by BDL. Additionally, online databases identified 49 potential protein targets for TBHQ. Finally, a biological target candidate (Keap1) was evaluated in a proof-of-concept in silico molecular docking assay, resulting in an interaction energy of -5.5491 kcal/mol, which was higher than RA839 and lower than monoethyl fumarate (compounds known to bind to Keap1). These findings suggest that TBHQ increases the survival of animals subjected to CCl4 intoxication or BDL, presumably by reducing hepatocellular damage, probably due to the interaction of TBHQ with Keap1.

Antioxidant and Protective Effect of Ocimum basilicum Seeds Extract on Renal Toxicity Induced by Carbon Tetrachloride in Rats.

Basil (Ocimum basilicum L.) is distributed worldwide and used in the food, pharmaceutical, and cosmetic industries. Most applications are for the herb basil, recently the basil seeds have also been used commercially; however, little is known about the nutritional and functional properties of the seeds. The present study aimed to investigate a possible protective effect of the methanol extract of O. basilicum seeds (MEOB), based on its phytochemical content, against kidney toxicity induced by CCl4 in adult rats. A single dose of CCl4 was used to induce oxidative stress in rats, which was demonstrated by a significant rise of serum enzyme markers. MEOB was administrated for 15 consecutive days (200 mg/kg body weight) to Wistar rats before CCl4 treatment and the effects on serum urea, creatinine, and uric acid, as well as the kidney superoxide dismutase, catalase, glutathione peroxidase, and glutathione activity and thiobarbituric acid reactive substances and protein carbonyl (PCO) levels were evaluated. In addition, histopathological examinations of kidneys were performed. In the positive control group, CCl4 induced an increase in serum biochemical parameters and triggered oxidative stress in the kidney. MEOB (200 mg/kg BW) resulted in significant reduction of CCl4-elevated levels of kidney markers, urea and creatinine, and a significant increase of uric acid compared with the CCl4-only group. In addition, MEOB pretreatment resulted in a significant reduction in lipid peroxidation and PCO levels in renal tissue compared with CCl4-exposed group. MEOB definitely could prevent the development of pathological changes in the kidneys. Overall, we conclude that MEOB is effective in protecting renal function from CCl4 toxicity.

Hepatoprotective effect of Indonesian propolis from Apis mellifera in carbon tetrachloride (CCl4) induced liver injury in mice

Abstract Propolis has been reported to have a broad spectrum of biological activities. The objective of this study was to investigate the hepatoprotective effect of propolis on liver injury induced by carbon tetrachloride (CCl4). Twenty-five adult mice were randomly divided into five groups of five. The normal group received distilled water; positive control group was given CCl4 at 2.8 mL/kg BW orally, other groups were given the same dose of CCl4, followed by oral propolis at 25, 50, and 100 mg/kg BW, respectively, for 14 days. On the 15th day, the mice were sacrificed for the measurements of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total protein, and albumin, as well as examination of liver histology. The data were analyzed using a one-way ANOVA and, subsequently, by Duncan’s Multiple Range Test. The results demonstrated that propolis had a hepatoprotective effect as shown by significant improvement of the biochemical parameters (p <0.05), which was confirmed by the liver histological analytical results. The most potent hepatoprotective effect was found after the dose of 100 mg/kg BW.

FREE RADICAL SCAVENGING ACTIVITY OF PLEUROTUS OSTREATUS AGAINST CCL4-INDUCED HEPATIC DAMAGE IN WISTER RATS

Objective: The present study was aimed to evaluate the anti-oxidant effect of Pleurotus ostreatus using in vitro and in vivo methodology. Methods: Methanolic extract of Pleurotus ostreatus (MEPO) was prepared by cold maceration technique. Rats weighing 150-200 g were divided into 6 groups of six each (n=6). Group I served as control (0.9% Nacl), group II was considered as the positive control, administered with CCl4 dissolved in olive oil (intraperitoneal injection at 0.5 mg/kg body weight), group III was given with CCl4+standard drug sylimarin (100 mg/kg body weight, p. o.), group IV, V and VI were administered with CCl4+MEPO at 200, 400 and 800 mg/kg orally for a period of 21 d. The liver injury was induced by the administration of CCl4 intraperitoneally with a single dose of CCl4 (0.5 mg/kg body weight) as a 1:1 (v/v) solution in olive oil. Results: In the Hydroxyl free radical scavenging assay, the IC50 values of the test extract was found to be 72.1±1.68 as compared with the standard drug (IC50-66.3±1.02), which was considered as significant (***p<0.001). In the reducing power assay, all the absorbance values were comparable to standard ascorbic acid and showed maximum reducing ability at the concentration of 500 µg/ml. In the in vivo anti-oxidant study, administration of MEPO to injury-induced rats at the dose of 400 and 800 mg/kg, significantly increased (**p<0.001) the levels of SOD and CAT, with a significant reduction in the MDA levels. Conclusion: The current study explored a potential source for anti-oxidants in the methanolic extract of Pleurotus ostreatus, thus seems to be protective to liver in CCl4-induced hepatic injury.

Grewia asiatica fruit extract-based kalari cheese for enhanced storage stability and functional value

The study aimed to develop a functional cheese using Grewia asiatica fruit extract as a novel additive and demonstrated its functional value in oxidative-stress-induced [single dose of CCl4 (1 mL/kg) given intraperitoneally] male Wistar rats. The kalari cheese was developed using buffalo milk containing different levels of the fruit extract (0.0, 0.5, 0.7, and 1.0 %) and an optimum level of 0.7 % was found sensorily. The products (0.0 and 0.7 %) were stored in a refrigerator for 4 weeks and evaluated for storage quality. The extract improved the lipid (TBARS and free-fatty acids) and microbial (total plate and psychrophilic counts) stability and sensory quality of the cheese. The extract-enriched cheese-based diet ameliorated the impact of oxidative stress in the rats during a 4-week feeding trial and significantly (P < 0.05) increased the levels of endogenous antioxidant enzymes (superoxide dismutase, glutathione peroxidase and catalase) and body weight whereas significantly (P < 0.05) reduced the levels of liver marker enzymes (ALT and AST), lipid peroxidation, and relative liver and spleen weights compared to the control. The extract improved the lipid and microbial stability of the cheese and the cheese-based diet ameliorated the effect of oxidative stress in rats, indicating the functional value of the developed product.

Effects of Andrographis paniculata on Carbon Tetrachloride (CCl4)-Mediated Renal Oxidative Damage in Rats

A herbal medicinal plant known as Andrographis paniculata, or "hempedu bumi," is recognised for its numerous medicinal properties and role in promoting community health. Despite its widespread use, the potential nephroprotective effects and underlying mechanism of action of Andrographis paniculata remain unexplored. To address this gap, the present study aimed to investigate the nephroprotective effects of Andrographis paniculata against renal oxidative damage induced by carbon tetrachloride (CCl4) in rats. Sprague-Dawley rats were pre-treated with Andrographis paniculata extract via gavage (100, 200, and 300 mg/kg b.w., respectively) once daily for 14 days, followed by two doses of CCl4 (1.2 ml/kg b.w.) on the 13th and 14th days. After two weeks, rats were sacrificed, and a nephroprotective analysis was performed. CCl4 administration at a dose of 1.2 ml/kg body weight resulted in oxidative stress in the renal system, as evidenced by elevated lipid peroxidation levels (TBARS). This oxidative stress was accompanied by a significant decrease in the activities of antioxidant enzymes and a depletion in the levels of reduced glutathione (p&lt;0.05). Histopathological examination confirmed the impairment of renal function. Andrographis paniculata significantly mitigated the majority of these alterations. Based on our research, the nephroprotective advantages of Andrographis paniculata can be attributed to its ability to act as an antioxidant and scavenge free radicals.

Economic Importance of Garcinia Kola: Evidences of Shielding Outcome Against Carbon Tetrachloride-Induced Kidney Toxicity in Experimental Models

Introduction: The aim of the study was to evaluate the hepatoprotective effect of aqueous extract of Garcinia kola) seeds on carbon tetrachloride (CCl4) induced liver damage in adult Wistar rats. Thirty adult Wistar rats weighing between 65-145g were randomly divided into six groups of 5 rats each. Group A (control) rats were orally administered with 5ml/kg body weight of distilled water daily for 2 weeks, group B rats were administered distilled water orally daily for 2 weeks (volume per body weight) and carbon tetrachloride (CCl4) 0.4 m1/kg intraperitonially as a single application on day 14 of the experiment. Group C rats were administered 100 mg of silymarin / kg body weight once daily for 2 weeks followed by a single dose of CCl4 (0.4 m1) on day 14 of the experiment. Group D rats were administered 400 mg aqueous extract bitter kola (Garcinia kola) / kg body weight orally once daily for 2 weeks. Group E and F rats were administered 400 mg and 200 mg aqueous extract bitter kola (Garcinia kola) seed) orally once daily for 2 weeks followed by a single dose of CCl4 (0.4 m1) on day 14 respectively. At the end of the experiment. Garcinia kola seed aqueous extract caused significant decrease in the aspartate aminotransferase (AST) levels in the serum of the extract treated groups. Histopathological examinations revealed distortion of histoarchitecture such as, sinusoidal congestion, necrosis, steatosis and fibrosis was observed in group B. The administration of aqueous extract of Garcinia kola seed remarkably inhibited histoarchitectural distortion induced by CCl4 administration. Effects of the extract at dose of 200mg/kg was comparable to the reference drug. Garcinia kola aqueous seed extract showed a remarkable hepatoprotective and antioxidant activity against CCl4 induced hepatotoxicity as observed from the serum marker enzymes and antioxidant levels in liver tissues. CCl4 induced a significant rise in AST, ALT, and ALP with an increase of superoxide dismutase (SOD), catalase (CAT) and reduction lipid peroxidation (MDA). Treatment of the rats with the extract significantly (p&lt;0.05) altered serum marker enzymes and antioxidant levels to near normal compared with CCl4- treated rats (group B). The activity of the extract at dose of 400mg/kg (group F) was comparable to the standard drug confirmed by histopathological examinations of liver sections. Conclusion: Garcinia kola aqueous extract has hepatoprotective and antioxidant properties against CCl4-induced hepatotoxicity in Wistar rats.

A comparative study on the hepatoprotective effect of selenium-nanoparticles and dates flesh extract on carbon tetrachloride induced liver damage in albino rats.

Exposure to environmental pollutants such as carbon tetrachloride (CCL4) causes liver damage. This study aimed to compare the ameliorative activity of the dates flesh extract (DFE) and selenium-nanoparticles (SeNPs) on CCL4-induced hepatotoxicity and if DFE could be a useful alternative supplement. Twenty-four male albino rats were enrolled and randomly divided into four equal groups (6 rats in each group): control group received only basal diet with no medications. Group II received CCL4 in a dose of 0.5 mg/kg intraperitoneal injection twice weekly for four weeks. Group III rats were pretreated with SeNPs in a dose of 2.5 mg/kg once a day orally three times/wk for four weeks alone then combined with the previously described dose of CCL4 for another four weeks. Group IV rats were pretreated with DFE in a dose of 8 ml of the aqueous extract/kg/d orally for four weeks alone then combined with the previously described dose of CCL4 for another four weeks. The liver damage was assessed by estimation of plasma concentration of albumin and enzymes activities of alanine aminotransferase and tissue genes expression. Liver oxidation levels were assessed by measuring the tissue concentration of the malondialdehyde, superoxide dismutase, and the total glutathione. Additionally, inflammatory mediators tumour necrosis factor--α and interleukin-6 were estimated. Detecting the liver's cellular structural damage was done by histopathological and immunohistochemical examination. This study suggests that CCL4-induced liver damage in rats can be protected by administration whether the costly SeNPs or the economical DFE.

Hepatoprotektif Ekstrak Batang Bajakah (Spatholobus littoralis Hassk) Induksi Tetraclorida (CCL4)

This study aims to explore the benefits of bajakah (Spatholobus littoralis Hassk) through the study of its utilization as a hepatoprotector. This research method is quantitative. Bajakah stem extract was made using 96% ethanol solvent using the maceration method. Analysis was carried out using liver histology preparations to determine the HAI (Histology Activity Index) Knodel Score. The treatment group consisted of 5 groups, namely, a) negative control (K1) treated with toxic doses of CCL4; b) positive control, namely vitamin E 46.8 mg/KgBW; c) P1 (treatment 1) was 750 mg/KgBW of Bajakah stem extract; d) P2 (treatment two) was 1200 mg/KgBW of pirated stem extract; e) P3 (treatment of three) was extract of the stem of Bajakah 1500 mg/KgBB. The results showed that the post hoc LSD test results in the P1 group had a histopathological picture of damaged liver, there was a significant difference with the K and P3 groups (p &lt;0.05), but there was no significant difference when compared with the P2 group. Meanwhile, the P3 group had significant differences when compared to all groups (p &lt;0.05). In conclusion, the administration of extract of pirated stems at a dose of 1,500 mg/Kg has a significant effect on the HAI value on the average number of hepatocytes with parenchymatous degeneration and necrosis after CCL4 induced&#x0D; Keywords: Bajakah, CCL4, Hepatoprotective&#x0D;

Exploration of Perturbed Liver Fibrosis-Related Factors and Collagen Type I in Animal Model of Non-Alcoholic Fatty Liver Disease.

To determine their involvement in the onset of the disease, we investigated the changing levels of liver fibrosis-related proteins, namely, type-I collagen, α-smooth muscle actin (α-SMA), and transforming growth factor β1 and β3 (TGF-β1, β3). The four groups of Sprague-Dawley (SD) rats were involved in the study, namely, (i) normal control group, (ii) high-fat diet group (HFD), (iii) carbon tetrachloride (CCl4) group, and (iv) NAFLD group (animal model) which were chosen at random. The NAFLD model received HFD combined with subcutaneous injection of small doses of CCl4. Histopathological examination confirmed extent of liver fibrosis, while other immunological and molecular methods were used to evaluate expression and distribution of α-SMA, type I collagen TGF-β1 and TGF-β3, at both m-RNA and protein levels. In contrast to the normal control group, the NAFLD group showed moderately elevated expressions of TGF-β1, α-SMA, and type I collagen, which was proportional on temporal scale of NAFLD persistence in the model (P < 0.05). In the early phage of NAFLD, enhancement in the mRNA transcripts and, henceforth, protein expression of TGF-β3 was observed. However, these were found to be downregulated in case of liver fibrosis (P < 0.05). This NAFLD rat model shows the histopathologic changes of human NAFLD and is suitable for the study of NAFLD pathogenesis. These findings suggest that type I collagen and the liver fibrosis-related factors TGF- β1, TGF- β3, and α-SMA may be significant contributors to NAFLD. Although NAFLD model is previously demonstrated by other researchers, our study is novel in terms of exploration of involvement of fibrosis-related factors and in particular aforementioned proteins at the early stage of NAFLD vis-à-vis dynamics of type-I collagen distribution.

Nicotinic acid attenuates experimental non-alcoholic steatohepatitis by inhibiting the NLRP3 inflammasome/pyroptosis pathway

Non-alcoholic steatohepatitis (NASH) is a global public health concern that may progress into fibrosis, cirrhosis, and liver cancer, with limited curative treatment options. While the nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome is closely linked to NASH progression, nicotinic acid (NA), a vitamin used for the treatment of dyslipidemia, is an emerging pharmaceutical treatment for hepatic steatosis and fibrosis. Here, we investigated pharmacological effects of NA on experimental NASH and whether NLRP3 inflammasome/pyroptosis inhibition is an associated mechanism of action. Rats were fed a high-fat sucrose diet supplemented with cholesterol and a low dose of CCl4. NA significantly reduced inflammation by decreasing the protein levels of tumor necrosis factor-alpha and nuclear factor kappa B. Moreover, NA inhibited the formation of NLRP3- apoptosis-associated speck-like protein containing caspase recruitment domain-Caspase-1, decreasing interleukin-1beta, interleukin-18, and gasdermin D protein. In addition, NA reduced tumor growth factor-beta, alpha-smooth muscle actin, and hepatic levels of collagen-1, consequently decreasing extracellular matrix synthesis. Our results indicate that NA can inhibit NASH progression and encourage further basic and clinical studies on the use of NA for the treatment of human NASH.

Assessment of active constituents and total flavonoids of Ammi majus plant extracts on immunological and kidney protective activities in mice

The Ammi majus (Kella) plant is frequently used to cure various health issues. This study aimed to investigate the most important phytochemical compounds of Kella seeds and evaluate the effect of its aqueous and alcoholic extracts on CCl4-induced nephrotoxicity in mice using a count of white blood cells (WBC) test and some biochemical markers for renal functions besides the histopathological study of kidney tissue. According to secondary metabolite analysis, the seeds contained tannins, alkaloids, saponins, flavonoids, and polyphenols at varying levels depending on the solvent utilized. Regarding total flavonoids, the alcoholic extract showed the highest concentration (193.2±25.7 mg/g) compared to the aqueous extract (176.3±19.2 mg/g). The WBC count test in the in vivo study showed an improved immune system by increasing the number of WBCs in mice treated with plant extracts and reducing the toxic effects of the CCl4 compound compared with a negative control group. In the positive control group (treated mice with CCl4), the biochemical analysis and histological study of the kidney sections revealed that CCl4 causes nephrotoxicity through increasing urea and creatinine concentrations in the blood and lowering total protein content, as well as the formation of necrotic tissue in the kidney sections. In contrast, these markers decreased in groups of mice treated with plant extracts, particularly alcoholic extract. In conclusion, the A. majus plant can improve kidney function during CCl4 doses. Consequently, it could be a promising treatment for nephrotoxicity caused by certain drugs. Keywords: Ammi majus; kidney markers; nephrotoxicity

Hepatotoxicity Induced by Carbon Tetrachloride in Experimental Model

The present study is the first attempt to evaluate the hepatotoxicity induced by carbon tetrachloride (CCl4) in experimental model. It poses a significant hazard to one's health. It is also one of the leading sources of toxicity in critical organs such as the lungs, kidneys, liver, and brain. Objective: To assess the hepatotoxicity of carbon tetra chloride in albino rats. Methods: The research was conducted at the Department of Zoology, University of Okara. The experiment was conducted at the animal home of the Department of Zoology, University of Okara. There were two groups created: a control group and an experimental group. The experimental group was treated with CCl4. The rats were fed 30% diluted carbon tetrachloride with normal saline as a control group to test the harmful effect on the liver profile. This was accomplished through a 12-day trial. Sampling or dissection was done after 12 days. Rats were dissected, and their liver was punctured to obtain a blood sample and organ collection. After sampling was taken by puncturing the Rats' liver, the samples were examined by a machine called Micro-Lab 300. Results: Histopathological studies also proved that the liver of rats was damaged. The hepatotoxic dose of CCl4 also raised the serum AST, ALP, ALT, and bilirubin levels. Total levels of AST, ALP, ALT, and Bilirubin were higher than usual, indicating that CCl4 has a toxic effect on the liver profile of rats. Conclusions: This study suggested that CCl4 induced toxicity in rat liver.

Efficacy of Saudi propolis and bee pollen in the reduction of oxidative stress induced with CCl4 in a testis mice model

Low testosterone levels are caused by alcoholism, cigarette smoking, and exposure to toxic chemicals. This work focused on investigating the activities of propolis (PE) and bee pollen (BPE) extracts in reducing the oxidative stress of carbon tetrachloride (CCl4) in male mice models. The 48 male Swiss Albino mice weighed 27.5 ± 2.5 g and were divided into: Group1: Control (-) received distilled water only through oral intubation; Group 2: Control (- -) received corn oil by intraperitoneal (i.p.) injection once a day; Group 3: Control (+) received a sublethal dose of CCL4 intraperitoneally the end of the experiment. Group 4: Stander treated with silymarin at a daily dose of 200 mg/kg orally. Group 5: The mice were given 8.4 mg/ kg bw of (PE) orally. Group 6: The mice were given (BPE) extract (140 mg/kg bw) orally. After five consecutive days of treatment, all mice had testis injury in all groups except G1& G2, by a single i.p injection of CCL4 at a dose of 0.5 mL/kg (bw; 20% v/v in corn oil). The result showed a significant increase in luteinizing, follicle-stimulating, and testosterone hormone levels in the serum and semen parameters in the groups treated with PE and BPE. The histological results showed the greatest improvements in testis structures in the BPE group, which was confirmed using (Bcl-2; immunohistochemistry). These results suggest an important role of the antioxidative effects of PE and BPE in the attenuation of CCl4 oxidative stress.