Carbon Tetrachloride-induced Liver Injury Research Articles

The Ameliorative Effects of Gadolinium Chloride on Carbon Tetrachloride Induced Liver Injury in Albino Rats through Suppressing Tgfb1 Expression

The Ameliorative Effects of Gadolinium Chloride on Carbon Tetrachloride Induced Liver Injury in Albino Rats through Suppressing Tgfb1 Expression

Liver-targeted delivery of asiatic acid nanostructured lipid carrier for the treatment of liver fibrosis

Liver fibrosis is a major global health concern. Management of chronic liver disease is severely restricted in clinics due to ineffective treatment approaches. However, a lack of targeted therapy may aggravate this condition. Asiatic acid (AA), a pentacyclic triterpenoid acid, can effectively protect the liver from hepatic disorders. However, the pharmaceutical application of AA is limited by low oral bioavailability and poor targeting efficiency. This study synthesized a novel liver-targeting material from PEG-SA, chemically linked to ursodeoxycholic acid (UA), and utilized it to modify AA nanostructured lipid carriers (UP-AA-NLC) with enhanced targeting and improved efficacy. The formulation of UP-AA-NLC was optimized via the Box–Behnken Experimental Design (BBD) and characterized by size, zeta potential, TEM, DSC, and XRD. Furthermore, in vitro antifibrotic activity and proliferation of AA and NLCs were assessed in LX-2 cells. The addition of UP-AA-NLC significantly stimulated the TGF-beta1-induced expression of α-SMA, FN1, and Col I α1. In vivo near-infrared fluorescence imaging and distribution trials in rats demonstrated that UP-AA-NLC could significantly improve oral absorption and liver-targeting efficiency. Oral UP-AA-NLC greatly alleviated carbon tetrachloride-induced liver injury and fibrosis in rats in a dosage-dependent manner, as reflected by serum biochemical parameters (AST, ALT, and ALB), histopathological features (H&E and Masson staining), and antioxidant activity parameters (SOD and MDA). Also, treatment with UP-AA-NLC lowered liver hydroxyproline levels, demonstrating a reduction of collagen accumulation in the fibrotic liver. Collectively, optimized UP-AA-NLC has potential application prospects in liver-targeted therapy and holds great promise as a drug delivery system for treating liver diseases.

Potential therapeutic effects of pomegranate peel on carbon tetrachloride induced liver injury in rat

Potential therapeutic effects of pomegranate peel on carbon tetrachloride induced liver injury in rat Abstract: Pomegranate (Punica granatum L.) peel, a by-product of juice processing industries was reported to contain a series of bioactive compounds. This study was undertaken to investigate the putative antioxidant activity of the dried pomegranate peel and their extract clinically in carbon tetrachloride (CCl4) induced liver damage in male rats was used. Intraperitoneal administration of CCl4 (2 ml/kg) to rats for 4 days resulted in significantly elevated (p≤0.05) serum levels of liver enzymes compared to controls. In consequence, significant elevation (p≤0.05) of malondialdehyde (MDA) and a lowered level of reduced glutathione (GSH) were observed. When rats with CCl4-induced hepatotoxicity were treated with the dried grape leaves and their water extract, the serum GOT, GPT and ALP levels reverted to near normal. Concurrently, the hepatic concentration of GSH was significantly increased (p≤0.05) and that of MDA significantly (p≤0.05) lowered when compared to CCl4 exposed untreated rats. Histopathological studies confirmed the hepatoprotective effect profounded by the extracts. These results suggest that pomegranate peel extract is able to significantly alleviate the hepatotoxicity induced by CCl4 and can protect liver against some pathological diseases. Keywords: pomegranate peel; liver fibrosis; hepatoprotective; antifibrotic effect; histopathology. التأثيرات العلاجية المحتملة لقشر الرمان على رباعي کلوريد کربون الکبد المستحث في الفئران د. هبة ابراهيم غمري قسم الاقتصاد المنزلى - کلية الاقتصاد المنزلى جامعة الملک خالد ، المملکة العربية السعودية تعتبر قشور الرمان منتج ثانوي من صناعات معالجة العصير والتى تحتوي على عدد کبير من المرکبات النشطة حيويا. وقد أجريت هذه الدراسة للبحث في النشاط المضاد للأکسدة المتوقع لقشر الرمان المجفف وتأثيراته العلاجية على أضرار الکبد المستحثة برابع کلوريد الکربون (CCl4) في ذکور الفئران. وقد أدت المعالجة بـ CCl4 (2مل / کغ) للفئران لمدة 4 أيام الى مستويات الدم المرتفعة (p≤0.05) من أنزيمات الکبد مقارنة مع الکنترول. ونتيجة لذلک ، لوحظ ارتفاع کبير (p≤0.05) بمرکب المالونالدهيد MDA (مؤشر أکسدة الدهون) ومستوى منخفض من الجلوتاثيون (GSH). وعندما عولجت الفئران المصابة بالسمية الکبدية الناتجة عن CCl4 بقشور الرمان المجففة ومستخلصاتها المائية ، عادت مستويات إنزيمات الکبد ( GOT وGPT و ALP ) إلى ما يقرب من المعدل الطبيعي. في الوقت نفسه وازدادت GSH بشکل ملحوظ ترکيز الکبد p≤0.05 وانخفضت مستويات MDA بشکل کبير (p≤0.05) بالمقارنة مع بالمجموعة المعاملة بـ CCl4. کما أکدت الدراسات التشريحية النسيجية أن کبد الأمعاء تأثرت إيجابيا عند المعاملة بالمستخلصات. وتشير هذه النتائج إلى أن مستخلص قشور الرمان قادر على التخفيف بشکل کبير من التسمم الکبدي الناجم عن CCl4 ، کما أن له تأثير وقائى کبير على الکبد من الإصابة ببعض الأضرار المرضية. الکلماتالمفتاحية: قشر الرمان - تليف الکبد – الکبد - تأثير المضاد الحيوي - التشريح المرضي.

Mitochondrial transplantation therapy inhibit carbon tetrachloride-induced liver injury through scavenging free radicals and protecting hepatocytes.

Carbon tetrachloride (CCl4)‐induced liver injury is predominantly caused by free radicals, in which mitochondrial function of hepatocytes is impaired, accompanying with the production of ROS and decreased ATP energy supply in animals intoxicated with CCl4. Here we explored a novel therapeutic approach, mitochondrial transplantation therapy, for treating the liver injury. The results showed that mitochondria entered hepatocytes through macropinocytosis pathway, and thereby cell viability was recovered in a concentration‐dependent manner. Mitochondrial therapy could increase ATP supply and reduce free radical damage. In liver injury model of mice, mitochondrial therapy significantly improved liver function and prevented tissue fibrogenesis. Transcriptomic data revealed that mitochondrial unfold protein response (UPRmt), a protective transcriptional response of mitochondria‐to‐nuclear retrograde signaling, would be triggered after mitochondrial administration. Then the anti‐oxidant genes were up‐regulated to scavenge free radicals. The mitochondrial function was rehabilitated through the transcriptional activation of respiratory chain enzyme and mitophage‐associated genes. The protective response re‐balanced the cellular homeostasis, and eventually enhanced stress resistance that is linked to cell survival. The efficacy of mitochondrial transplantation therapy in the animals would suggest a novel approach for treating liver injury caused by toxins.

Activation of Hepatic Stellate Cells Requires Dissociation of E-Cadherin–Containing Adherens Junctions with Hepatocytes

Hepatic stellate cells (HSCs) are resident mesenchymal cells in the space of Disse interposed between liver sinusoidal endothelial cells and hepatocytes. Thorn-like microprojections, or spines, project out from the cell surface of HSCs, crossing the space of Disse, to establish adherens junctions with neighboring hepatocytes. Although HSC activation is initiated largely from stimulation by adjacent cells, isolated HSCs also activate spontaneously in primary culture on plastic. Therefore, other unknown HSC-initiating factors apart from paracrine stimuli may promote activation. The dissociation of adherens junctions between HSCs and hepatocytes as an activating signal for HSCs was explored, establishing epithelial cadherin (E-cadherin) as an adhesion molecule linking hepatocytes and HSCs. Invivo, following carbon tetrachloride-induced liver injury, HSCs lost their spines and dissociated from adherens junctions in the early stages of injury, and were subsequently activated along with an increase in YAP/TAZ expression. After abrogation of liver injury, HSCs reconstructed their spines and adherens junctions. Invitro, reconstitution of E-cadherin-containing adherens junctions by forced E-cadherin expression quiesced HSCs and suppressed TAZ expression. Additionally, increase of TAZ expression leading to the activation of HSCs by autocrine stimulation of transforming growth factor-β, was revealed as a mechanism of spontaneous activation. Thus, we have uncovered a critical event required for HSC activation through enhanced TAZ-mediated mechanotransduction after the loss of adherens junctions between HSCs and hepatocytes.

Apolipoprotein A4 regulates the immune response in carbon tetrachloride-induced chronic liver injury in mice

This article explores the role of ApoA4 in a CCl4-induced chronic liver injury (CLI) mouse model. C57BL/6J mice (WT) and ApoA4 knock-out (KO) mice were divided into CCl4 CLI (WT-CCl4 and KO-CCl4) and olive oil solvent control groups (WT-Veh and KO-Veh). Some of the KO-CCl4 mice were additionally treated with recombinant mouse ApoA4 and primary mouse T lymphocyte injections. After 6 weeks, histological analyses, biochemical and superoxide dismutase (SOD) and malondialdehyde (MDA) assays, flow cytometry of immune cells and qRT-PCR analyses were performed. KO mice after treatment with CCl4 showed reduced hepatic SOD and enhanced serum MDA activities leading to worsening liver injury and fibrosis compared with WT-CCl4, accompanied by enhanced hepatic alpha smooth muscle actin (α-SMA), tissue inhibitor of metalloproteinases-1 (TIMP-1) and collagen type I alpha 1 chain (COL1A1) transcriptions, elevated macrophage M1 levels, enhanced tumor necrosis factor-alpha (TNF-α), Interleukin 6 (IL-6) and C-C Motif Chemokine Ligand 5 (CCL5), but reduced Interleukin 10 (IL-10), monocyte chemotactic protein 1 (MCP-1), C-C Motif Chemokine Receptor 2 (CCR2), C-X3-C Motif Chemokine Receptor 1 (CX3CR1) and C-X-C Motif Chemokine Ligand 9 (CXCL9) transcription, as well as reduced CD3+, CD4+ and CD8+ T cell percentages in hepatic tissue, blood cells and spleen. In addition, CD11b+CD115+, CD11b+/Ly6Chigh, CD11b+/LyC6- and CD11b+/Ly6Cint cells were enhanced, which partly reversed by ApoA4 protein and T cell injections. In conclusion, we propose that ApoA4 might be involved in liver protection via inhibiting fibrotic mediators and inflammatory cytokines, suppression of pro-inflammatory hepatic M1 cell invasion and regulation of CD8+ T and CD4+ T lymphocytes.

Qing-Yu-Mu, an Herbal Formula, Reduces Hepatic Oxidative Stress in Rats Fed a High-Frying Oil Diet and Ameliorates Carbon Tetrachloride-Induced Liver Injury.

Qing-Yu-Mu (QYM) is an herbal formula used to prevent and treat liver disease in Taiwan. In this study, the hepatoprotective effects of QYM were evaluated in two experimental models. First, rats were fed a high-frying oil (FO) diet containing 1.25% QYM for 5 weeks to investigate effects of QYM on hepatic oxidative stress and antioxidant enzyme activities. Then, protective effects of QYM on carbon tetrachloride (CCl4)-induced chronic liver injury were evaluated. Results show that QYM treatment reduced FO diet-induced hepatic lipid peroxidation and reactive oxygen species levels and increased glutathione (GSH) S-transferase activity. A higher reduced GSH/oxidized GSH (GSSG) ratio was observed after QYM treatment. Furthermore, QYM ameliorated CCl4-induced liver injury by reducing the activity of plasma alanine aminotransferase and histological lesions in the liver. QYM also increased the level of hepatic GSH and activities of GSH peroxidase and superoxide dismutase. Finally, chlorogenic acid, chrysophanol, and apigenin were found to be present in relative abundance in QYM. Results show that QYM may exhibit a hepatoprotective effect by reducing oxidative stress and increasing antioxidant activity in the liver.

Carbon Dots from Paeoniae Radix Alba Carbonisata: Hepatoprotective Effect.

IntroductionThe charcoal processed product of Paeoniae Radix Alba (PRA), PRA Carbonisata (PRAC), has long been used for its hepatoprotective effects. However, the material basis and mechanism of action of PRAC remain unclear.AimTo explore the hepatoprotective effects of Paeoniae Radix Alba Carbonisata-derived carbon dots (PRAC-CDs).MethodsPRAC-CDs were characterized using transmission electron microscopy, high-resolution transmission electron microscopy, ultraviolet, fluorescence, Fourier transform infrared and X-ray photoelectron spectroscopy, X-ray diffraction, and high-performance liquid chromatography. The hepatoprotective effect of PRAC-CDs was evaluated and confirmed using the classic carbon tetrachloride acute liver injury model.ResultsPRAC-CDs averaged 1.0–2.4 nm in size and exhibited a quantum yield of 5.34% at a maximum excitation wavelength of 320 nm and emission at 411 nm. PRAC-CDs can reduce the ALT and AST levels of mice with carbon tetrachloride-induced acute liver injury and have a mitigating effect on the rise in TBA and TBIL. More interestingly, PRAC-CDs can significantly reduce MDA and increase SOD levels, demonstrating that PRAC-CDs can improve the body’s ability to scavenge oxygen free radicals and inhibit free radical-induced liver cell lipid peroxidation, thereby preventing liver cell damage.ConclusionThese results demonstrate the remarkable hepatoprotective effects of PRAC-CDs against carbon tetrachloride-induced acute liver injury, which provide new insights into potential biomedical and healthcare applications of CDs.

Morphological changes in parenchymal organs of laboratory animals in acute effect of carbon tetrachloride

Introduction. Among noninfectious diseases, one of the leading places belongs to pathologies caused by the action of industrial toxicants. In this regard, the search for drugs for the prevention and treatment of various intoxications is one of the priority tasks of public health. Treatment of intoxication with drugs with antioxidant and antihypoxic activity is more effective. The aim of this study was to investigate morphological changes in parenchymal organs (liver, kidneys, and pancreas) under acute exposure to high doses of carbon tetrachloride and to evaluate the effectiveness of their correction with a new composition of oxymethyluracil with ascorbic acid. Material and methods. The composition of 5-hydroxy-6-methyluracil with ascorbic acid was first synthesized at the Ufa Institute of Chemistry, Ufa Federal Research Center, RAS. The preventive effect of the new drug was studied in comparison with “Heptor” on the carbon tetrachloride-induced liver injury model. Morphological studies of the liver, pancreas, and kidneys of laboratory animals were carried out. Results. Studies have shown the prophylactic administration of new composition of 5-hydroxy-6-methyluracil with ascorbic acid to have a protective effect on the structure of parenchymal organs in acute carbon tetrachloride intoxication, comparable to the drug “Heptor” (possibly superior). However, reparative properties were observed only in the drug “Heptor”. Conclusion. Comparative estimation of morphological changes in parenchymal organs under acute exposure to high doses of carbon tetrachloride indicates the protective effect of prophylactic administration of the composition of 5-hydroxy-6-methyluracil with ascorbic acid, comparable to the drug “Heptor” (possibly superior).

Kadsura heteroclita stem ethanol extract protects against carbon tetrachloride-induced liver injury in mice via suppression of oxidative stress, inflammation, and apoptosis

Ethnopharmacological relevanceKadsura heteroclita stem (KHS) is a well-known hepatoprotective Tujia ethnomedicine (folk named Xuetong), has long been used for the prevention and treatment of hepatitis and liver diseases.Aim of the study: To explore the protective effects of KHS against carbon tetrachloride (CCl4)-induced liver injury and the underlying mechanism, particularly antioxidative, anti-inflammatory, and anti-apoptotic potentials. Materials and methodsThe acute toxicity of KHS was measured by the method of maximum tolerated dose (MTD). Liver injury in mice was induced by intraperitoneal injection of 25% carbon tetrachloride (olive oil solubilization) 2 times every week. After modeling, mice in KHS groups were treated with KHS at 100, 200, 400 mg/kg/d, mice in positive control group were treated with bifendate (30 mg/kg/d), and mice in normal and model groups were given ultrapure water. After 4 weeks of treatment, blood of mice was taken from the orbital venous plexus before mice euthanized, the liver, spleen, and thymus of mice were weighed by dissecting the abdominal cavity after mice euthanized. Moreover, the liver of mice was selected for histological examination. The alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in mice serum were measured using the automatic biochemical analyzer. The levels of superoxide dismutase (SOD), myeloperoxidase (MPO), malondialdehyde (MDA), glutathione peroxidase (GPX-2), tumor necrosis factor (TNF-α), interleukin-6 (IL-6), interleukin-10 (IL-10), Bcl-2-associated X (Bax), B-cell lymphoma-2 (Bcl-2), Caspase-3, and Caspase-8 in mice liver were measured by Elisa kits. Furthermore, the protein expression of Bcl-2 and Bax in mice liver tissue was detected by Western blot. ResultsThe MTD of KHS was determined to be 26 g/kg in both sexes of mice. Treatment with KHS dose-dependently protected the liver and other main organs against CCl4-induced liver injury in mice. The ALT and AST levels in mice liver were significantly reduced after treatment with KHS at the dose of 100, 200, and 400 mg/kg. In addition, the liver histopathological analyses revealed that KHS markedly alleviated inflammatory cell infiltration, hepatic fibrosis, hepatocyte ballooning, necrosis and severe apoptosis of hepatocytes induced by CCl4. Further assay indicated that KHS significantly suppressed the production of MDA and MPO, while markedly increased the level of SOD and GPx-2. The TNF-α and IL-6 level in mice liver tissue were decreased by KHS, whereas the IL-10 level was increased. KHS also inhibited hepatocyte apoptosis by significantly reducing the expression of Bax, Caspase-3, Caspase-8, as well as increasing the expression of Bcl-2. Besides, the Western blot results strongly demonstrated that KHS inhibited hepatocyte apoptosis, as evidenced by reducing the expression of Bax protein and increasing the expression of Bcl-2 protein in liver injury tissues. ConclusionsThis research firstly clarified that KHS has a significant protective effect against CCl4-induced liver injury, which might be closely related to alleviating oxidative stress, reducing inflammatory response, and inhibiting hepatocyte apoptosis.

Anwulignan Alleviates Carbon Tetrachloride-Induced Acute Liver Injury in Mice

Acute liver injury is a serious threat to human health. Complementary therapy including a traditional Chinese herb has been used for the prevention and treatment of liver injuries. Schisandrae sphenantherae fructus (Schisandra) is the mature dry fruit of Schisandra sphenanthera Rehd. et Wils. Wuzhi capsule, a preparation containing Schisandra and its main component anwulignan, is used to treat hepatitis and hepatic insufficiency caused by viruses and drugs in the clinic. However, to date, there has been little study to reveal the effect of anwulignan in the protection of the liver. Therefore, in this study, we hypothesized that anwulignan could protect carbon tetrachloride (CCl4)-induced acute liver injury in mice. Anwulignan was shown to reduce significantly the liver index, decrease liver histopathological injury, decrease the serum level of aspartate aminotransferase and alanine aminotransferase, increase the activities of superoxide dismutase (SOD) and glutathione peroxidase, reduce liver malondialdehyde content, and downregulate the expression levels of interleukin (IL)-6, IL-1β, and tumor necrosis factor-α in the liver tissue, as well as the protein expression levels of receptor-interacting serine/threonine-protein kinase 1 (RIPK1), RIPK3, and phosphorylated mixed lineage kinase domain-like protein. All these results suggest that anwulignan can alleviate the CCl4-induced acute liver injury in mice, which may be related to its antioxidant, anti-inflammation, and inhibition of liver cell necroptosis effects.

Pecan pericarp extract protects against carbon tetrachloride-induced liver injury through oxidative mechanism in rats.

The purpose of this study was to quantify the proanthocyanidin content of pecan (Carya illinoinensis) pericarp extract (PPE) and to assess its useful impacts against carbon tetrachloride (CCl4)-induced hepatotoxicity. Rats were randomly divided into four groups: Group 1: received intraperitoneal injection of saline solution, Group 2: was injected with PPE (25mg/kg body weight) for 10 consecutive days, Group 3: received CCl4 (0.5ml/kg, subcutaneous injection), Group 4: was coadministred with PPE+CCl4. The CCl4 was administered every 3days during 10days. Results revealed the presence of a high amount of total proanthocyanidins in the PPE (81.01±0.21mg TAE.g-1DW). CCl4 injection induced significant reductions in hepatic antioxidants but increased hepatic lipid peroxidation (LPO) as well as serum injury biomarkers. However, cotreatment with PPE significantly (P<0.05) inverted CCl4-induced increase in plasma alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and lactate dehydrogenase activities, respectively to 74%, 77%, 60%, and 82% compared with CCl4 group. No significant toxic effects were observed following treatment with plant extract alone. PPE cotreatment also decreased significant (P<0.05) the hepatic malondialdehyde formation (21%) and enhanced the liver catalase activity (107%) in CCl4-intoxicated rats. The histopathological examination showed inflammatory infiltration and degenerative changes in the hepatic tissue following CCl4 injection. The hepatoprotective activity of PPE against CCl4 exposure was supported by the maintenance of structural integrity of liver histopathology. In conclusion, the current study illustrated that PPE pretreatment significantly improved all examined parameters, restored the hepatic architecture and successfully alleviates oxidative damage induced by CCl4 intoxication.

Effect of fermented red radish (Raphanus sativus L) in carbon tetrachloride-induced liver injury in rats

Effect of fermented red radish (Raphanus sativus L) in carbon tetrachloride-induced liver injury in rats

Mycelia Extract from Spent Ganoderma Lucidum Bagasse Substrate Ameliorates Carbon Tetrachloride-Induced Acute Liver Injury in Mice Through Inhibiting Oxidative Stress

Sugarcane bagasse has been utilized by Ganoderma lucidum in high value, but this also produces tons of spent substrate. For comprehensive exploitation of sugarcane bagasse, this study was to evaluate the potential of mycelia extract from spent Ganoderma substrate (MESGS). In vivo assays, MESGS could alleviate acute liver injury induced by carbon tetrachloride in male Institute of Cancer Research (ICR) mice without toxicity. Analysis revealed that MESGS could effectively relieve the histopathological lesions of liver tissue. There are significant tendencies to descend for the levels of aspartate aminotransferase and alanine aminotransferase with the increasing doses of MESGS. Similarly, plasma creatinine levels of mice treating with MESGS were restored to 11.2~13.24 µmol/L which approached to 9.48 µmol/L of the normal control group. MESGS could mitigate symptoms of liver index raising and splenomegaly. It could be known that after acute liver injury, the mice with MESGS -treating had less side effects on liver, kidney and spleen. Meanwhile, the decreased activity of superoxide dismutase and increased malonaldehyde level were inhibited by MESGS treatment. In vitro assays, MESGS also exhibited strong antioxidant in a dose dependent manner by radicals scavenging activity with 1,1-Diphenyl-2-picrylhydrazyl, hydroxyl and 3-ethylbenzthiazoline-6-sulfonic acid diammonium salt. Furthermore, MESGS is rich in polysaccharide (22.13%) and polyphenolics (2.02%). These findings demonstrated that the MESGS displayed certain hepatoprotective effect on the acute liver injury by inhibiting oxidative stress. Therefore, the MESGS could be beneficial for drug development against chemical injury. It provides research basis for further improving high value conversion of sugarcane bagasse.

Byakangelicin protects against carbon tetrachloride-induced liver injury and fibrosis in mice.

Liver fibrosis is a disease caused by long‐term damage that is related to a number of factors. The current research on the treatment of liver fibrosis mainly focuses on the activation of hepatic stellate cell, in addition to protecting liver cells. byakangelicin has certain anti‐inflammatory ability, but its effect on liver fibrosis is unclear. This study aims to explore whether byakangelicin plays a role in the development of liver fibrosis and to explore its mechanism. We determined that byakangelicin has a certain ability to resist fibrosis and reduce liver cell damage in a model of carbon tetrachloride–induced liver fibrosis in mice. Thereafter, we performed further verification in vitro. The signalling pathways of two important pro‐fibrotic cytokines, transforming growth factor‐β and platelet‐derived growth factor, were studied. Results showed that byakangelicin can inhibit related pathways. According to the hepatoprotective effect of byakangelicin observed in animal experiments, we studied the effect of byakangelicin on 4‐HNE–induced hepatocyte (HepG2) apoptosis and explored its related pathways. The results showed that byakangelicin could attenuate 4‐HNE–induced hepatocyte apoptosis via inhibiting ASK‐1/JNK signalling. In conclusion, byakangelicin could improve carbon tetrachloride–induced liver fibrosis and liver injury by inhibiting hepatic stellate cell proliferation and activation and suppressing hepatocyte apoptosis.

Investigation of benfotiamine’s protective effects on liver tissue in experimental carbon tetrachloride induced liver injury

Purpose: In this study, we aimed to investigate the protective effects of benfotiamine on experimental liver injury caused by carbon tetrachloride (CCl4).Materials and Methods: In this study, 30 male Wistar albino rats were used. Rats were equally divided into 5 groups. No application was made to control group. The CCl4 group was injected i.p with1ml/kg CCl4:olive oil (1:2) mixture on the 1st and 8th days, and the CCl4+benfotiamine group was treated i.p with 1 ml/kg CCl4: olive oil (1:2) mixture twice on the 1st and 8th days and orally with 70 mg/kg/day benfotiamine. To the benfotiamine group, 70 mg/kg/day benfotiamine was given orally for 14 days. To the olive oil group, 2 ml/kg olive oil was given i.p. on 1st and 8th days. Finally, rats were decapitated. Liver tissues were removed and paraffin blocks were prepared. Tissues were stored at –80 oC for malonaldeyhde (MDA) assay.Results: There were no significant differences between the control, benfotiamine and olive oil groups. Compared with the control group, there was a significant increase in MDA, apoptosis and bax immunoreactivity in CCl4 group. Compared with the CCl4 group, there was a significant decrease in MDA, apoptosis and bax immunoreactivity in the CCl4+benfotiamine group.Conclusion: CCl4 increases MDA, apoptosis and bax immunoreactivity, and benfotiamine, given as treatment, reduces these parameters.

Structural characterization of phosphorylated Pleurotus ostreatus polysaccharide and its hepatoprotective effect on carbon tetrachloride-induced liver injury in mice

This study aimed to explore the basic structural features of phosphorylated Pleurotus ostreatus polysaccharide (PPOP) and study the protective effect of PPOP on liver injury induced by carbon tetrachloride in male Kunming mice. The phosphorylated polysaccharide was prepared from the natural polysaccharide extracted from Pleurotus ostreatus (POP). The structures of PPOP and POP were characterized by FT-IR, ESEM spectroscopy, and Congo red test. Chemical composition analysis revealed that PPOP was mainly composed of rhamnose, galacturonic acid, and xylose in a molar ratio of 0.10: 1.98: 1.00. Structural analysis indicated that PPOP had multi-strand structure and the absorption peaks of PO and P-O-C. Furthermore, animal experiments showed that the hepatoprotective effect of PPOP against liver injury was reflected by decreasing the levels of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, total cholesterol, trilaurin, and low-density lipoprotein cholesterol in the serum, increasing the content of high-density lipoprotein cholesterol and albumin in blood, reducing the content of malondialdehyde and promoting the activity of antioxidant enzymes in liver. PPOP exhibited stronger hepatoprotective effect and antioxidant activity in vivo than POP. The final results indicated that PPOP could be used in the treatment of chemical-induced hepatotoxicity based on the above biological research.

Hepatoprotection of pine nut polysaccharide via NRF2/ARE/MKP1/JNK signaling pathways against carbon tetrachloride-induced liver injury in mice

Previously, we obtained a purified polysaccharide (PNP40c-1) from Pinus koraiensis pine nut and reported its protective effect on carbon tetrachloride (CCl4)-induced liver injury in vitro. The object of this study is to investigate its hepatoprotective activity in vivo and elucidate the mechanism underlying the hepatoprotection. PNP40c-1 effectively prevented the accumulation of serum liver injury biomarkers including alanine aminotransferase, aspartate aminotransferase, alkaline phpsphatase and total bilirubin stimulated by CCl4. The pathological changes in PNP40c-1-treated mice livers were also markedly ameliorated. Results showed that PNP40c-1 suppressed the production of reactive oxygen species (ROS) and lipid peroxidation, upregulated Nrf2/ARE pathway and enhanced the antioxidant capacity of hepatocytes. Furthermore, the reaction between Nrf2 and ARE promoted the generation of Mkp1, which inhibited the activation of JNK induced by CCl4, and suppressed hepatocytes apoptosis by regulating the protein expression of Bax, cleaved-Caspase-3 and Bcl2, exerting hepatoprotective activity. Taken together, upregulation of Nrf2/ARE pathway and suppression of JNK activation via Nrf2/ARE/Mkp1/JNK signaling pathways are the main mechanisms underlying the hepatoprotective effect of PNP40c-1 against CCl4-induced mice liver injury. These results indicated that PNP40c-1 has potential to serve as a hepatoprotective agent against chemical induced hepatotoxicity.

Anoectochilus roxburghii polysaccharide prevents carbon tetrachloride-induced liver injury in mice by metabolomic analysis

Anoectochilus roxburghii (Wall.) Lindl, a traditional Chinese medicine, is used for the effective treatment of liver disease in China. Anoectochilus roxburghii polysaccharide (ARPT) is an important constituent of Anoectochilus roxburghii. ARPT exerts a hepatoprotective effect and contributes directly to the therapeutic benefit of Anoectochilus roxburghii. However, the hepatoprotective mechanism of ARPT requires further elucidation. The present study was designed to assess the effects and underlying mechanism of ARPT when used to pretreat carbon tetrachloride (CCl4)-induced liver injury in mice. Mice were randomly divided into three groups: control group (no ARPT treatment or liver injury), model group (liver injury induced with CCl4), and the ARPT group (ARPT pretreatment followed by liver injury). A metabolomic method, based on liquid chromatography combined with mass spectrometry (LC-MS) and pattern recognition analysis, was applied. The data were analyzed with principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA), to determine differentiating metabolites in the serum and liver tissue between the experimental groups. The PCA and OPLS-DA scores plots of the serum and liver tissue samples based on the LC-MS data showed a clear separation between the control and liver injury model group, while the ARPT-treated group showed a trend of close with the control. There were eleven metabolites [PS(O-18:0/0:0), phosphocholine, phenylalanine, hippuric acid, α-ketoisovaleric acid, metyrosine, leucinic acid, ketoleucine, Cer(d18:1/19:0), α-kamlolenic acid, and 4-formyl indole] were identified as candidate biomarkers in the serum samples, eight such metabolites (valine, phosphohydroxypyruvic acid, phosphocholine, ornithine, indole, xanthine, uridine, and glucose 6-phosphate) were found in the liver tissue samples, and one metabolite (phosphocholine) was observed in both the serum and liver tissue samples. These endogenous metabolites are considered to be in response to the hepatoprotective effects of ARPT and are involved in amino acid metabolism, lipid metabolism, gut bacteria metabolism, energy metabolism, and the methylation pathway. These findings suggest that ARPT mitigates the metabolic effect of CCl4-induced hepatotoxicity in mice by affecting at least five different pathways. LC-MS-based metabolomics provides a powerful approach for identifying potential biomarkers and for elucidating the protective mechanisms of traditional Chinese medicines against disease.

Indole-3-carbinol alleviates carbon tetrachloride-induced liver injury by inhibiting inflammatory response and regulating lipid metabolism

The hepatoprotective effects of indole-3-carbinol (I3C), one of the byproducts of the breakdown of glucosinolate glucobrassicin, were examined in mice with carbon tetrachloride (CCl4)-induced hepatotoxicity. I3C was administered orally at a dose of 50 or 100 mg/kg to C57BL/6 mice for 3 consecutive days with or without a subsequent single oral administration of CCl4 (1.5 mL/kg). I3C treatment significantly suppressed the increases in alanine aminotransferase and aspartate aminotransferase levels in the sera of CCl4-injured mice. In addition, I3C treatment markedly suppressed CCl4-induced hepatic inflammation, by suppressing the activation of macrophages and Kupffer cells in the liver. To examine the effects of I3C on lipid accumulation, 3T3-L1 pre-adipocytes were differentiated with I3C or vehicle control. I3C treatment reduced lipid accumulation, which appeared to be associated with the downregulation of adipogenic transcription factors, such as CCAAT/enhancer-binding protein α, peroxisome proliferator-activated receptor γ, and lipid accumulation-related genes, including adipocyte protein-2. Taken together, the present study suggest that I3C treatment reduces hepatocyte death and lipid accumulation in hepatocytes in CCl4-induced hepatic injury with concurrent suppression of macrophages/Kupffer cells, which was further evidenced by I3C-treated pre-adipocytes in vitro, through downregulation of lipid metabolism.