CCl4-induced Acute Liver Damage Research Articles

Synthesis and preclinical evaluation of a 89Zr-labelled human single chain antibody for non-invasive detection of hepatic myofibroblasts in acute liver injury

Synaptophysin is expressed on fibrogenic hepatic myofibroblasts. C1–3 is a single chain human antibody (scAb) that binds specifically to synaptophysin on hepatic myofibroblasts, providing a targeting vector for novel in vivo imaging agents of chronic liver disease. C1–3 and a negative control scAb, CSBD9, were radiolabelled with zirconium-89 via desferrioxamine chelation to enable non-invasive molecular imaging with positron emission tomography (PET). DFO-scAb conjugates were characterised by gel electrophoresis (SDS-PAGE) and MALDI-TOF spectrometry, and 89Zr-labelled with high radiolabelling efficiency (99%). [89Zr]Zr-DFO-C1–3 exhibited high in vitro stability (> 99%) in mouse and human sera over 3 days at 25 and 37 °C. Activated hepatic myofibroblasts incubated with [89Zr]Zr-DFO-C1–3 displayed significantly higher internalised activity (59.46%, P = 0.001) compared to the [89Zr]Zr-DFO-CSBD9 control, indicating synaptophysin-mediated uptake and high binding specificity of [89Zr]Zr-DFO-C1–3. Mice with CCl4-induced acute liver damage exhibited significantly higher liver uptake of [89Zr]Zr-DFO-C1–3, compared to controls, confirmed by both Cerenkov imaging and ex vivo gamma counting (4.41 ± 0.19%ID/g, P < 0.0001). CCl4-induced liver damage and the number of hepatic myofibroblasts was confirmed by αSMA staining of liver sections. These findings indicate that [89Zr]Zr-DFO-C1–3 has promising utility as a PET imaging agent for non-invasive detection of hepatic myofibroblasts following acute liver injury.

Structural characteristics and ameliorative effect of a polysaccharide from Corbicula fluminea industrial distillate against acute liver injury induced by CCl4 in mice

Corbicula fluminea distillate as an important industrial by-product of C. fluminea during steaming process is rich in amino acids, proteins and polysaccharides, showing potential hepatoprotective effect. In this study, a polysaccharide (CFDP) was obtained from C. fluminea distillate by three-phase partitioning combined with (NH4)2SO4 precipitation at a saturation of 60 %. The structural characteristics, antioxidant activity in vitro, and hepatoprotection against mice CCl4-induced acute liver damage of CFDP were studied. Results demonstrated that CFDP was a water-soluble homogenous polysaccharide predominantly comprising glucose (>98 %), with a weight-average molecular weight of 1.4 × 107 Da, and exhibiting potent antioxidant benefits in vitro. CFDP had a backbone of (1 → 4)-α-d-glucopyranosyl (Glcp) and a small amount of (1 → 4, 6)-α-D-Glcp. The branch formed at C-6 comprised by (1→)-α-D-Glcp and (1→)-α-D-N-acetylglucosamine. CFDP possessed excellent hepatoprotective activity against acute liver damage caused by CCl4 in mice, mainly by ameliorating weight reduction and organ injures, alleviating hepatic function and serum lipid metabolism, suppressing oxidative stress and inflammatory responses, as directly verified by histopathological examination. Moreover, CFDP improved gut microbiota by up-regulating the relative abundance of total bacteria and probiotics such as Firmicutes, Bacteroidete, Rumminococcaceae, Lactobacillaceae, accompanied by promoting short chain fatty acid production. Therefore, our findings indicated that CFDP can be developed as a healthy food supplement for the prevention of chemical livery injury.

Protective effect of Euphorbia thymifolia and Euphorbia hirta against carbon tetrachloride-induced hepatotoxicity in Wistar rats

Objectives The present study aims to investigate the protective effect of Euphorbia thymifolia and Euphorbia hirta extracts on in vitro antioxidant activity and in vivo analysis on hepatic marker enzyme levels and histopathological changes in the liver of carbon tetrachloride (CCl4) induced hepatotoxicity rats. Materials and methods This study includes 42 adult male Albino Wistar rats randomly divided into seven treatment groups, including control (basal diet, G1), CCl4-induced single dose (1.5 ml/kg, i.p.) as the negative control (G2), G1 supplemented with 300 mg/kg of ethanol extract of E. thymifolia (G3) and E. hirta (G4), G2 supplemented with 300 mg/kg of ethanol extract of E. thymifolia (G5), E. hirta (G6), and silymarin (25 mg/kg b.w.) used as a standard drug (G7) for 21-days experimental period. Results The ethanolic extracts of E. thymifolia and E. hirta exhibited potential in vitro antioxidant activity in a dose-dependent manner (25 μg/ml, 50 μg/ml, 100 μg/ml, 200 μg/ml and 250 μg/ml). Oxidative stress caused by CCl4-induced the liver damage, including changes in liver marker enzymes (aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase), enzymatic (superoxide dismutase and catalase), non-enzymatic antioxidants (lipid peroxides and glutathione) and hepatocellular alterations such as hydropic degeneration, irregular hepatocytes, and distention of the vein. Administration of E. thymifolia and E. hirta significantly (p < 0.05) restored the enzyme activity along with the histology of the liver. Conclusion The results from the current study demonstrate that E. thymifolia and E. hirta have the property of restoring hepatic redox capacity and antioxidant activities against CCl4-induced acute liver damage.

Tamarix dioica (Ghaz) Protective Potential in the Carbon Tetrachloride-Induced Hepatotoxicity Animal Model

Introduction: Hepatic diseases remain the leading cause of death worldwide. Despite overall advancements in health care, mortality due to hepatic diseases is constantly growing. More than 2 million people globally are estimated to die each year from liver diseases, and current treatment offers little for its management. Thus, it is essential to find more effective and less toxic pharmaceutical alternatives for the treatment of liver diseases.Aims &amp; Objectives: Tamarix dioica, a shrub broadly used in herbal medicine for the treatment and prevention of various diseases. The current study was designed to analyze the hepatoprotective effect of T. dioica in BALB?cmice against CCl4-induced acute liver damage.Place and duration of study: The study was conducted in NIH, Islamabad, Pakistan, for six months in 2016-2017. Material &amp; Methods: For in vivo evaluation, the animals (n= 42) were randomly divided into seven groups (n=6), three control (i.e. Group, I or normal control, group II or induction control received 0.9% normal saline orally, and Group III or positive control received silymarin 100 mg/kg per oral), and four treatment groups (i.e. IV, V,VI and VII were treatedwith oral T.dioica 200 mg/kg/day, 300mg/kg/day methanol extract, 200mg/kg/day and 300mg/kg/day of aqueous extracts respectively for six days, followed by intraperitoneal administration of CCl4 on the seventh day. The blood samples were collected for analysis of LFTs, and hepatic tissue was taken for histological analysis. Data was analyzed using SPSS version 16, one-way ANOVA with Duncan’s Multiple Range Test (DMRT).Results: CCl4 induction in Group 2 resulted in severe hepatic derangement manifested as highly elevated mean LFTs (ALT 7245.56, AST 3292.11, ALP 340.09 U/L, bilirubin 4.64 mg/dl) as compared to healthy controls (ALT 38.97, AST 50.20, ALP 57.17 U/L, bilirubin 1.25 mg/dl: (Group 1) levels p&lt;0.001. Pretreatment with different extracts of T.dioica for 6 days before CCl4 administration produced varying degrees of hepatoprotection. 300mg/kg aqueous extract T.dioica(Group7) prevented damage with maximal hepatoprotection, reduced LFTs (ALT: 339.95 , AST: 242.90 , ALP: 116.86 U/L, bilirubin: 1.38 mg/dl) and normalized liver histology as compared to Group 2 and standard drug silymarin 100mg/kg, (ALT: 6483.23, AST: 2567.69, ALP: 272.19 U/L, bilirubin: 2.84 mg/dl: Group 3) p&lt;0.001. Lesser hepatoprotection was provided by T.dioica aqueous extract 200mg/kg (ALT: 439.93, AST: 367.87, ALP: 180.62 U/L bilirubin: 1.53 mg/dl: Group VI) and least by 300mg/kg &amp; 200mg/kg methanolic extracts Groups V &amp; IV (ALT: 6338.06, 6443.91, AST: 2800.81, 3012.34, ALP: 242, 248 U/L &amp; bilirubin: 2.82 &amp; 3.62 mg/dl) respectively. Further, no drug-induced toxicity symptoms were observed 24 hours after administration of the high dose oral T. dioica 2000 mg/kg/body weight aqueous and methanolic extracts were administered.Conclusion: Pretreatment with T. dioica extracts especially 300mg/kg aqueous extract reduced acute CCl4-mediated liver damage, ameliorated histopathological as well as biochemical parameters and was free of toxicity in 2000mg/kg /body weight dose in the mice experimental model. T. dioica has potential in hepatoprotective drug research.

The novel nanocomplexes containing deoxycholic acid-grafted chitosan and oleanolic acid displays the hepatoprotective effect against CCl4-induced liver injury in vivo

Chemical liver injury threatens seriously human health, along with the shortage of efficiency and low-toxicity drugs. Herein, the novel oral nanocomplexes composed of deoxycholic acid-grafted chitosan and oleanolic acid were constructed to reverse the CCl4-induced acute liver damage in mice. Results indicated core-shell nanocomplexes, maintained by the hydrophobic interaction between deoxycholic acid and oleanolic acid, could be dissociated in the intestine. Notably, the nanocomplexes possessed superior hepatoprotective effect in vivo, possibly due to the synergistic effect between grafted chitosan and oleanolic acid. Mechanism investigations suggested that nanocomplexes reversed CCl4-induced liver injury through improving hepatic antioxidant capacity via NrF2/Keap1 pathway and regulating inflammation response via NF-κB signaling pathway. The novel oral nanocomplexes represent an effective strategy for chemical liver injury therapy.

Design, Preparation, and Characterization of Dioscin Nanosuspensions and Evaluation of Their Protective Effect against Carbon Tetrachloride-Induced Acute Liver Injury in Mice.

The purpose of this study was to prepare a dioscin nanosuspension (Dio-NS) that has a better distance and high solubility for oral administration and to evaluate its hepatoprotective effects. Optimal primary manufacture parameters, including shear time, shear speed, emulation temperature, pressure, and cycles of homogenization, were determined by single-factor experiments. The concentrations of dioscin, SDS, and soybean lecithin were optimized using the central composite design-response surface method, and their effects on the mean particle size (MPS) and particle size distribution of Dio-NS were investigated. Characterization of the Dio-NS formulations included examinations of the surface morphology and physical status of dioscin in Dio-NS, the stability of Dio-NS at different temperatures, in vitro solubility, and liver protective effect in vivo. Under optimal conditions, Dio-NS had an MPS of 106.72 nm, polydispersity index of 0.221, and zeta potential of −34.27 mV. Furthermore, the proportion of dioscin in Dio-NS was approximately 21.26%. The observation of particles with a spherical shape and the disappearance of crystalline peaks indicated that the physical and chemical properties of Dio-NS were altered. Furthermore, we observed that the dissolution of Dio-NS was superior to that of a physical mixture and Dio-GZF. Moreover, Dio-NS was demonstrated to have a protective effect against CCl4-induced acute liver damage in mice that was equivalent to that of silymarin (a positive control drug) at the same dose. The good hepatoprotective effect of our Dio-NS preparation can provide a theoretical basis for investigating its absorption mechanisms in the body.

A strategy for quality evaluation of salt-treated Apocyni Veneti Folium and discovery of efficacy-associated markers by fingerprint-activity relationship modeling

In this study, a fingerprint-activity relationship between chemical fingerprints and hepatoprotective activity was established to evaluate the quality of salt-treated Apocyni Veneti Folium (AVF). Characteristic fingerprints of AVF samples exposed to different concentrations of salt were generated by ultrafast liquid chromatography tandem triple time-of-flight mass/mass spectrometry (UFLC-Triple TOF-MS/MS), and a similarity analysis was performed based on common characteristic peaks by hierarchical clustering analysis (HCA). Then, the hepatoprotective activity of AVF against CCl4-induced acute liver damage in mice was investigated by assessing biochemical markers and histopathology, which showed that a high dose of AVF exposed to low levels of salt stress produced a marked amelioration of hepatic damage compared with the other salt-treated AVF. Finally, fingerprint-activity relationship modeling, which was capable of discovering the bioactive markers used in the quality evaluation, was investigated by the chemical fingerprints and the hepatoprotective activities utilizing multivariate statistical analysis, gray correlation analysis (GCA) and bivariate correlation analysis (BCA). The results showed that the accumulation of polyphenols, such as flavonoids and phenolic acids, in AVF subjected to low levels of salt stress could result in the effective scavenging of free radicals. Therefore, the present study may provide a powerful strategy to holistically evaluate the quality of salt-treated AVF in combination with chemical fingerprint and bioactivity evaluation.

Liver Targeting Albumin-Coated Silybin-Phospholipid Particles Prepared by Nab™ Technology for Improving Treatment Effect of Acute Liver Damage in Intravenous Administration.

In this study, a novel human serum albumin nanoparticle loading silybin-phospholipid complex (SLNPs) was developed for liver targeting after intravenous administration. The preparation of the drug delivery system consisted of two steps; initially, a silybin-phospholipid complex (SLC) was produced to improve the lipophilicity of SLB to then achieve enhanced encapsulation of SLB in albumin nanoparticles. FT-IR and XRD analysis confirmed the successful formation of SLC. The complex ratio of SLC in the first step was 99.6%. The encapsulation efficiency and drug loading of SLNPs in the second step were 96.2% and 5.6%, respectively. SLNPs were spherical and well-dispersed, with a zeta potential of approximately - 10mV, and a mean particle size around 200nm. An in vivo tissue distribution experiment and a pharmacodynamic experiment showed that, compared with SLB solution, SLNPs had an improved SLB accumulation in the liver. The hepatoprotective effect of SLNPs on CCl4-induced acute liver damage was evaluated. CCl4-damaged mice showed an increased enzymatic activity of ALT and AST; however, enzyme levels returned to near-normal levels in high-dose SLNP-treated mice. As SLNPs combine the enhanced oil solubility of SLC and the passive targeting of albumin nanoparticles, they possess great potential for the treatment of acute liver damage.

Andrographolide derivative as STAT3 inhibitor that protects acute liver damage in mice

Sustained activation of the Janus kinase-signal transducers and activators of transcription (JAK-STAT) pathway contributed to the progression of cancer and liver diseases. STAT3 signaling inhibitor has been extensively investigated for pharmacological use. We synthesized a series of andrographolide derivatives, and characterized their activity against STAT3 signaling pathway both in vitro and in the CCl4-induced acute liver damage mice model. Among these derivatives, compound 24 effectively inhibited phosphorylation and dimerization of STAT3 but not its DNA binding activity. Compound 24 significantly ameliorated carbon tetrachloride-induced acute liver damage in vivo without changing mice body weight. Treatment with 24 attenuated hepatic pathologic damage and promoted hepatic proliferation and activation of STAT3. Compound 24 inhibited elevated expression of α-smooth muscle actin and serum pro-inflammatory cytokines downstream of STAT3 but not those factors that are regulated by NF-κB or SMADs. In summary, our results suggest that compound 24 may serve as a potential therapeutic agent for the treatment of hepatic damage or a liver protection agent via regulating STAT3 activation.

Comparison of the Hepatoprotective Effects of Four Endemic Cirsium Species Extracts from Taiwan on CCl₄-Induced Acute Liver Damage in C57BL/6 Mice.

Species of Cirsium (Asteraceae family) have been used in folk hepatoprotective medicine in Taiwan. We collected four Cirsium species—including the aerial part of Cirsium arisanense (CAH), the aerial part of Cirsium kawakamii (CKH), the flower part of Cirsium japonicum DC. var. australe (CJF), and Cirsii Herba (CH)—and then made extractions from them with 70% methanol. We compared the antioxidant contents and activities of these four Cirsium species extracts by a spectrophotometric method and high-performance liquid chromatography–photodiode array detector (HPLC-DAD). We further evaluated the hepatoprotective effects of these extracts on CCl4-induced acute liver damage in C57BL/6 mice. The present study found CAH possesses the highest antioxidant activity among the four Cirsium species, and these antioxidant activities are closely related to phenylpropanoid glycoside (PPG) contents. The extracts decreased serum ALT and AST levels elevated by injection with 0.2% CCl4. However, only CJF and CH decreased hepatic necrosis. Silibinin decreased serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels and hepatic necrosis caused by CCl4. CJF and CH restored the activities of hepatic antioxidant enzymes and decreased hepatic malondialdehyde (MDA) levels. CJF further restored the expression of hepatic antioxidant enzymes including Cu/Zn-superoxide dismutase (Cu/Zn-SOD), Mn-superoxide dismutase (Mn-SOD), and glutathione S-transferase (GST) proteins. HPLC chromatogram indicated that CKH, CJF, and CH contained silibinin diastereomers (α and β). Only CJF contained diosmetin. Hence, the hepatoprotective mechanism of CJF against CCl4-induced acute liver damage might be involved in restoring the activities and protein expression of the hepatic antioxidant defense system and inhibiting hepatic inflammation, and these hepatoprotective effects are related to the contents of silibinin diastereomers and diosmetin.

Characterization, antioxidant activity and hepatoprotective effect of purple sweetpotato polysaccharides

In this study, three kinds of polysaccharides (named PSWP, PSAP-1 and PSAP-2) were successively isolated from purple sweetpotato tubers by hot water, 0.5M and 2M sodium hydroxide solutions. The characterization, in vitro antioxidant activity and in vivo hepatoprotective effect of these polysaccharides were investigated. Results indicated that PSWP, PSAP-1 and PSAP-2 were all β-type polysaccharides containing different contents of uronic acid, protein and polyphenol. Both PSWP and PSAP-1 were composed by arabinose, glucose and galactose, whereas PSAP-2 was consisted of arabinose, rhamnose and glucose. All the polysaccharides exhibited moderate 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and reducing power. As compared with tetrachloromethane (CCl4) treatment group, mice administrated with PSWP, PSAP-1 and PSAP-2 exhibited decreased levels of serum enzymes (alkaline phosphatase, alanine transaminase and aspartate transaminase) and hepatic lipid peroxidation, whereas increased levels of hepatic antioxidant enzymes (superoxide dismutase, catalase and glutathione peroxidase) and non-enzymatic antioxidant (glutathione). Notably, PSWP displayed stronger antioxidant activity and hepatoprotective effect than PSAP-1 and PSAP-2. The hepatoprotective effect of PSWP was comparable to positive standard of silymarin. Our results suggested polysaccharides from purple sweetpotato possessed potential antioxidant activity and protective effect against CCl4-induced acute liver damage.

Protective Effects of Melatonin on CCl4-induced Acute Liver Damage and Testicular Toxicity in Rats

Effects of melatonin against carbon tetrachloride-induced acute liver and testicular toxicity in rats as well as the possible mechanisms were investigated in the present study. Rats were acclimatized and the serum levels of alanine aminotransferase, aspartate aminotransferase and reduced glutathione were measured. The malondialdehyde, superoxide dismutase, and reduced glutathione concentrations in the hepatic homogenate, and histopathological changes in rat liver and testes were also determined. Membrane potential was analysed with flow cytometry. The expression level of telomerase and caspase-3 were determined by reverse transcriptase polymerase chain reaction. Melatonin increased the content of serum reduced glutathione in rats with acute liver injury induced by carbon tetrachloride. The extent of malondialdehyde formation was reduced; superoxide dismutase activity and the reduced glutathione contents were increased in the hepatic homogenate in melatonin groups. Histopathological examination of rat liver and testicular sections confirmed the biochemical observations. Melatonin significantly restored the plasma membrane potential and decreased mRNA level of telomerase and caspase-3. These results indicated that melatonin exerted a significant protective effect against acute hepatotoxicity induced by carbon tetrachloride in rats, which might be due to free radical scavenging, inhibition of lipid peroxidation, increasing antioxidant activity as well as restoration plasma membrane potential. The expression of telomerase and caspase-3 might partially contribute to the protective effects on carbon tetrachloride-induced acute liver damage. Reduced glutathione content might also partially contribute to the protective effects of melatonin on the oxidative damage caused by carbon tetrachloride in rat testis.

Hepatoprotective effect of methyl ferulic acid against carbon tetrachloride-induced acute liver injury in rats.

The present study aimed to investigate the hepatoprotective effects of methyl ferulic acid (MFA) against oxidative stress and apoptosis in acute liver injury induced by carbon tetrachloride (CCl4) in rats, as well as the underlying mechanisms. Sprague Dawley rats were treated with CCl4 after oral administration of MFA (25, 50, and 100 mg/kg) or dimethyl diphenyl bicarboxylate (200 mg/kg) for 7 days. The hepatoprotective effects of MFA were determined by analyzing serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities as well as changes of oxidant parameters. Histopathological analysis was performed to determine the degree of hepatic injury. The mechanisms were investigated by detecting the levels of NADPH oxidase (NOX) trans-membrane subunit NOX4, its ligand p22phox, as well as caspase3, cleaved caspase3, B-cell lymphoma (Bcl)-2, Bcl-2-associated X protein (Bax), tumor necrosis factor (TNF)-α, interleukin (IL)-1, reactive oxygen species (ROS), thiobarbituric acid-reactive substances (TBARS), total anti-oxidant capacity (TAC), phosphorylated J-Jun N-terminal kinase (p-JNK) and p-p38 mitogen-activated protein kinase (MAPK) using semi-quantitative polymerase chain reaction, western blot analysis and colorimetric assays. MFA treatment significantly decreased serum enzymatic activities of ALT and AST. MFA markedly increased activities of liver superoxide dismutase, catalase and glutathione peroxidase, and reduced the malondialdehyde concentration. Histopathological examination demonstrated that MFA reduced lipid degeneration, cytoplasmic vacuolization, necrosis and inflammatory cell infiltration in the liversof CCl4-treated rats. MFA treatment markedly inhibited the expression of inflammatory factors TNF-α and IL-1β. Mechanistic study revealed that MFA decreased the TAC and the levels of ROS and TBARS. Furthermore, MFA treatment led to a reduction of the mRNA and protein expression of NOX4 and p22phox, as well as the protein levels of caspase3, cleaved caspase-3 and Bax, and an upregulation of p-JNK, p-p38 MAPK and Bcl-2 proteins in the liver. The present study demonstrated that MFA has hepatoprotective effects against CCl4-induced acute liver damage. MFA has anti-oxidant, anti-inflammatory and anti-apoptotic activities and was able to modulate the NOX4/p22phox/ROS-JNK/p38 MAPK signaling pathway.

Experimental approach to IGF-1 therapy in CCl4-induced acute liver damage in healthy controls and mice with partial IGF-1 deficiency

BackgroundCell necrosis, oxidative damage, and fibrogenesis are involved in cirrhosis development, a condition in which insulin-like growth factor 1 (IGF-1) levels are diminished. This study evaluates whether the exogenous administration of low doses of IGF-1 can induce hepatoprotection in acute carbon tetrachloride (CCl4)-induced liver damage compared to healthy controls (Wt Igf+/+). Additionally, the impact of IGF-1 deficiency on a damaged liver was investigated in mice with a partial deficit of this hormone (Hz Igf1+/−).MethodsThree groups of 25 ± 5-week-old healthy male mice (Wt Igf+/+) were included in the protocol: untreated controls (Wt). Controls that received CCl4 (Wt + CCl4) and Wt + CCl4 were treated subcutaneously with IGF-1 (2 µg/100 g body weight/day) for 10 days (Wt + CCl4 + IGF1). In parallel, three IGF-1-deficient mice (Hz Igf1+/−) groups were studied: untreated Hz, Hz + CCl4, and Hz + CCl4 + IGF-1. Microarray and real-time quantitative polymerase chain reaction (RT-qPCR) analyses, serum aminotransferases levels, liver histology, and malondialdehyde (MDA) levels were assessed at the end of the treatment in all groups. All data represent mean ± SEM.ResultsAn altered gene coding expression pattern for proteins of the extracellular matrix, fibrosis, and cellular protection were found, as compared to healthy controls, in which IGF-1 therapy normalized in the series including healthy mice. Liver histology showed that Wt + CCl4 + IGF1 mice had less oxidative damage, fibrosis, lymphocytic infiltrate, and cellular changes when compared to the Wt + CCl4. Moreover, there was a correlation between MDA levels and the histological damage score (Pearson’s r = 0.858). In the IGF-1-deficient mice series, similar findings were identified, denoting a much more vulnerable hepatic parenchyma.ConclusionsIGF1 treatment improved the biochemistry, histology, and genetic expression of pro-regenerative and cytoprotective factors in both series (healthy and IGF-1-deficient mice) with acute liver damage, suggesting that low doses of IGF-1, in acute liver damage, could be a feasible therapeutic option.

Hepatoprotective effect against CCl4-induced acute liver damage in mice and High-performance liquid chromatography mass spectrometric method for analysis of the constituents of extract of Rubus crataegifolius

This study is an attempt to evaluate the hepatoprotective activity of Rubus Crataegifolius against carbon tetrachloride-induced liver injury in mice. 70% ethanolic, ethyl acetate and n-BuOH extract of R. crataegifolius were administered daily for 14 days in experimental animals before they were treated with CCl4. The hepatoprotective activity of the extracts in this study was compared with the reference drug silymarin. A high-performance liquid chromatography mass spectrometric (HPLC-EIS-MS/MS) method was developed for the determination of the constituents of the extracts. According to the data of HPLC-EIS-MS/MS, the chemical structures of the largely 14 constituents of R. crataegifolius were identified online without time-consuming isolation. Ethyl acetate extracts of R. crataegifolius showed strong antioxidant activities and significant protective effect against acute hepatotoxicity induced by CCl4. According to the data of HPLC-EIS-MS/MS, Oleanic acid, Phlorizin dehydrate and Quercetin-3-rhamnoside are considered as the main hepatoprotective factor in ethyl acetate extract.

Hepatoprotective activity of chrysin is mediated through TNF-α in chemically-induced acute liver damage: An in vivo study and molecular modeling.

Chrysin (5,7-dihydroxyflavone) is a naturally occurring flavonoid present at high levels in honey, propolis and numerous plant extracts. Chrysin is known to have hepatoprotective activity, however, the mechanisms by which it exerts this effect remain unclear. In the present study, the effects of chrysin in carbon tetrachloride (CCl4)-induced acute liver damage were investigated and the results used to infer a possible mechanism behind chrysin's hepatoprotective activity. Prior to an intraperitoneal injection of CCl4 (1 ml/kg) to induce acute liver damage, chrysin (50 mg/kg) was administered orally to mice for 7 days. The positive control group was given 50 mg/kg standardized silymarin, a well-studied hepatoprotective flavonoid. Twenty-four h following CCl4 administration, an increase in the activity levels of serum aspartate-amino-transferase and alanine-amino-transferase was found. This was accompanied by extended centrilobular necrosis, steatosis and an altered hepatocyte ultrastructure. In addition, CCl4-induced acute hepatotoxicity was associated with an increase in hepatic tumor necrosis factor-α (TNF-α) and α-smooth muscle actin (α-SMA) protein expression, which was significantly decreased in the livers of mice pre-treated with chrysin (P<0.001), similar to the results of the silymarin pre-treated group (P<0.001). Treatment with chrysin prior to CCl4 exposure significantly reduced the activity of enzymes used as biochemical markers of poor liver function compared with the group which did not receive pre-treatment (P<0.001). In addition, the results of histopathological and electron microscopy liver examination showed chrysin pre-treatment reduced the effects of CCl4 treatment. Molecular modeling results demonstrated that the hepatoprotective activity of chrysin is mediated through TNF-α, as it reduces soluble TNF-α generation via blocking TNF-α-converting enzyme activity. In conclusion, the results of the present study suggest that inflammatory pathways are activated in CCl4-induced acute liver damage, which are ameliorated by chrysin pre-treatment. This indicates that chrysin is a potent hepatoprotective agent, similarly to silymarin at the same dose, which has the potential to be a viable alternative to conventional hepatoprotective treatments.

Protective effect of a polysaccharide from Anoectochilus roxburghii against carbon tetrachloride-induced acute liver injury in mice

Ethnopharmacological relevanceAnoectochilus roxburghii (Wall.) Lindl. is traditionally used for the treatment of various types of chronic and acute hepatitis in China. Considering that Anoectochilus roxburghii polysaccharide (ARPT) is the main constituent of Anoectochilus roxburghii, the present study was designed to investigate the hepatoprotective effect of ARPT and its possible mechanism in carbon tetrachloride (CCl4)-induced mice. Material and methodsThe hepatoprotective activity of ARPT (150, 300 and 500mg/kg) were investigated on CCl4-induced acute liver damage in mice. The activities of alanine transaminase (ALT), aspartate transaminase (AST) were determined in serum. The hepatic levels of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) were measured in liver homogenates. The levels of cytochrome P450 sub family 2E1 (CYP2E1), tumor necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1β), interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein-2 (MIP-2), KC (Murine IL-8 ortholog), transforming growth factor-beta1 (TGF-β1), Bcl-2 and Bax were measured by quantitative real-time polymerase chain reaction (qRT-PCR). The expressions of CYP2E1, nuclear factor-kappa B (NF-κB) p65 and caspase-3 were evaluated by western blot assays. The hepatic levels of TNF-α, IL-6, MIP-2 and TGF-β1 were measured by enzyme-linked immunosorbent assay (ELISA). Furthermore, histopathological observation and terminal-deoxynucleoitidyl transferase mediated nick end labeling assay (TUNEL) were carried out on the separated livers of mice. ResultsARPT significantly decreased serum ALT and AST activities, hepatic MDA level, and markedly enhanced antioxidant enzyme (SOD, CAT and GSH-Px) activities and GSH level in hepatic tissue, in a dose-dependent manner, when compared to the model group. Histopathological observation revealed the hepatoprotective effect of ARPT against the damage. Furthermore, ARPT remarkably inhibited CYP2E1 mRNA expression, decreased NF-κB p65 expression and therefore to prevent the secretion of pro-inflammatory cytokines (TNF-α and IL-6) and chemokines (MCP-1, MIP-2 and KC), suppressed TGF-β1 expression and hepatocytes apoptosis. Moreover, ARPT could prevent DNA fragmentation based on TUNEL assay results. ConclusionThese findings suggested that ARPT possessed hepatoprotective effect against CCl4-induced hepatotoxicity in mice and the action might in part be through reducing oxidative stress, inflammation and apoptosis.

Dicranostiga leptopodu (Maxim.) Fedde extracts attenuated CCl4-induced acute liver damage in mice through increasing anti-oxidative enzyme activity to improve mitochondrial function

Dicranostiga Leptodu (Maxim.) fedde (DLF), a poppy plant, has been reported have many benefits and medicinal properties, including free radicals scavenging and detoxifying. However, the protective effect of DLF extracts against carbon tetrachloride (CCl4)-induced damage in mice liver has not been elucidated. Here, we demonstrated that DLF extracts attenuated CCl4-induced liver damage in mice through increasing anti-oxidative enzyme activity to improve mitochondrial function. In this study, the mice liver damage evoked by CCl4 was marked by morphology changes, significant rise in lipid peroxidation, as well as alterations of mitochondrial respiratory function. Interestingly, pretreatment with DLF extracts attenuated CCl4-induced morphological damage and increasing of lipid peroxidation in mice liver. Additionally, DLF extracts improved mitochondrial function by preventing the disruption of respiratory chain and suppression of mitochondrial Na+K+-ATPase and Ca2+-ATPase activity. Furthermore, administration with DLF extracts elevated superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) levels and maintained the balance of redox status. This results showed that toxic protection effect of DLF extracts on mice liver is mediated by improving mitochondrial respiratory function and keeping the balance of redox status, which suggesting that DLF extracts could be used as potential toxic protection agent for the liver against hepatotoxic agent.

Antioxidant and Hepatoprotective Activities of Mycelia Selenium Polysaccharide by Hypsizigus marmoreus SK-02.

This work was designed to investigate the characteristic properties (bond types and monosaccharide compositions), and hepatoprotective effects on carbon tetrachloride (CCl4)-induced liver damage of mycelia selenium polysaccharides (MSPS) separated and purified from Hypsizigus marmoreus SK-02. Characteristic analysis of MSPS showed the selenium content (70.15μg/g) in mycelia. The antioxidant activities in vitro demonstrated that MSPS had potential effects on scavenging reactive oxygen species and enhancing the reducing power. The treatment of MSPS for CCl4-induced animal experiment demonstrated that the MSPS could reduce the levels of malondiadehyde (MDA), lipid peroxide (LPO), glutamic oxalacetic transaminase (AST), and glutamic-pyruvic transaminase (ALT) activities and improve the levels of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), total cholesterol (TC), and triglyceride (TG) in serum/liver homogenate against CCl4-induced injures. Findings presented in this study clearly demonstrated that MSPS might be suitable for functional foods and natural drugs in preventing the CCl4-induced acute liver damage.

Purification, characterization and hepatoprotective activities of mycelia zinc polysaccharides by Pleurotus djamor

This study was designed to investigate the physicochemical properties (molecular weights, bond types and monosaccharide compositions), antioxidant activities, and hepatoprotective effects on carbon tetrachloride (CCl4)-induced acute liver damage of mycelia zinc polysaccharides (MZPSs) and its major fractions (MZPS-1, -2 and -3) separated from Pleurotus djamor. In vitro assays, the MZPS-3 demonstrated relatively strong antioxidant activities in dose-dependent manners. For in vivo hepatoprotective activities, administration of MZPS-3 at 800mg/kg significantly decreased the levels of AST, ALT, MDA and LPO, remarkably increased the levels of TC, TG and ALB, and prominently restored the activities of SOD, GSH-Px, CAT and T-AOC in serum/liver homogenate against CCl4-induced injures. Findings presented in this study clearly demonstrated that MZPSs, especially MZPS-3, might be suitable for functional foods and natural drugs in preventing the CCl4-induced acute liver damage.