Novel D-galactosamine-induced cynomolgus monkey model of acute liver failure

AIM To establish a simplified, reproducible D-galactosamineinduced cynomolgus monkey model of acute liver failure having an appropriate treatment window. METHODS Sixteen cynomolgus monkeys were randomly dividedinto four groups(A, B, C and D) after intracranial pressure(ICP) sensor implantation. D-galactosamine at 0.3, 0.25, 0.20 + 0.05(24 h interval), and 0.20 g/kg body weight, respectively, was injected via the small saphenous vein. Vital signs, ICP, biochemical indices, and inflammatory factors were recorded at 0, 12, 24, 36, 48, 72, 96, and 120 h after D-galactosamine administration. Progression of clinical manifestations, survival times, and results of H&E staining, TUNEL, and Masson staining were recorded. RESULTS Cynomolgus monkeys developed different degrees of debilitation, loss of appetite, and jaundice after D-galactosamine administration. Survival times of groups A, B, and C were 56 ± 8.7 h, 95 ± 5.5 h, and 99 ± 2.2 h, respectively, and in group D all monkeys survived the 144-h observation period except for one, which died at 136 h. Blood levels of ALT, AST, CK, LDH, TBi L, Cr, BUN, and ammonia, prothrombin time, ICP, endotoxin, and inflammatory markers [(tumor necrosis factor(TNF)-α, interleukin(IL)-1β, and IL-6)] significantly increased compared with baseline values in different groups(P < 0.05). Pathological results showed obvious liver cell necrosis that was positively correlated with the dose of D-galactosamine.CONCLUSION We successfully established a simplified, reproducible D-galactosamine-induced cynomolgus monkey model of acute liver failure, and the single or divided dosage of 0.25 g/kg is optimal for creating this model.

Sinapic acid ameliorates D-galactosamine/lipopolysaccharideinduced fulminant hepatitis in rats:Role of nuclear factor erythroidrelated factor 2/heme oxygenase-1 pathways

BACKGROUND Sinapic acid(SA)has been shown to have various pharmacological properties such as antioxidant,antifibrotic,anti-inflammatory,and anticancer activities.Its mechanism of action is dependent upon its ability to curb free radical production and protect against oxidative stress-induced tissue injuries.AIM To study the hepatoprotective effects of SA against lipopolysaccharide(LPS)/Dgalactosamine(D-GalN)-induced acute liver failure(ALF)in rats.METHODS Experimental ALF was induced with an intraperitoneal(i.p.)administration of 8μg LPS and 800 mg/kg D-GalN in normal saline.SA was administered orally once daily starting 7 d before LPS/D-GalN treatment.RESULTS Data showed that SA ameliorates acute liver dysfunction,decreases serum levels of alanine transaminase(ALT),and aspartate aminotransferase(AST),as well as malondialdehyde(MDA)and NO levels in ALF model rats.However,pretreatment with SA(20 mg/kg and 40 mg/kg)reduced nuclear factor kappalight-chain-enhancer of activated B cells(NF-κB)activation and levels of inflammatory cytokines(tumor necrosis factor-αand interleukin 6).Also,SA increased the activity of the nuclear factor erythroid-related factor 2/heme oxygenase-1(Nrf2/HO-1)signaling pathway.CONCLUSION In conclusion,SA offers significant protection against LPS/D-GalN-induced ALF in rats by upregulating Nrf2/HO-1 and downregulating NF-κB.

Protective Effects of Dimethyl-4,4'-Dimethoxy-5,6,5',6'-Dimethylene Dioxybiphenyl-2,2'-Dicarboxylate on Damages of Isolated Rat Hepatocytes Induced by Carbon Tetrachloride and D-galactosamine

The protective effect of biphenyl dimethyl dicarboxylate (DDB) on chemically induced damages was studied in isolated suspended rat hepatocytes. The experimental results showed that DDB (200μg/106 cells) efficiently protected the hepatocytes against carbon tetrachloride (CC14 10 mrnol.L-1) and D-galactosamine (1 mmol.L-1) induced damages. Membranal lipid peroxidation (malondialdehyde, MDA formation) and glutamic pyruvic transaminase (GPT) release from the hepatocytes were markedly decreased. The damage of the cell surfaces of the hepatocytes were also reduced as seen under a scanning electron microscope (SEM). Pretreatment with DDB (300 mg-kg-1) orally ameliorated the reduction of liver glycogen and blood glucose caused by ip injection of D-galactosamine (800 mg-kg-1) in mice. When normal rats were given DDB 300 mg-kg-1 once daily for 10 d, the free ribosomal protein and RNA in the liver increased significantly. These results indicate that DDB is of beneficial effects on both damaged and normal hepatocytes.

Carvacrol suppresses the expression of inflammatory marker genes in D-galactosamine-hepatotoxic rats

Objective:To unravel the mechanism of anti-inflammatory activity of carvacrol in D-galactosamine(D-GalN)-induced hepatotoxic rats.Methods:The mRNA and protein expression levels of tumor necrosis factor-alpha(TNF-α).interleukin-6[IL-6).inducible nitric oxide synthase(iNOS),cyelooxy genase-2(COX-2) and nuclear faclor kappa-B(NF-κB) were assayed by semi-quantitative reverse transcriptase polymerase chain reaction(RTPCR) and western blot analysis.Results:We found that the mRNA and protein expressions of TNF-α. IL-6,iNOS,COX-2 and NF-κB were significanlly up-regulated in D-galactosamine induced hepatotoxic rats and treatment with carvacrol significantly down-regulated the expressions of these genes showing the mechanism behind the anti-inflammatory activity of carvacrol. Conclusions:All above results reveal that the carvacrol well known anti-inflammatory activities in D-galactosamine induced hepatotoxic rats.

Inhibiting the expression of hepatocyte nuclear factor 4 alpha attenuates lipopolysaccharide/ D-galactosamine-induced fulminant hepatic failure in mice

BACKGROUND: Hepatocyte nuclear factor 4 alpha (HNF4α) plays an important role in regulating cytokine-induced inflammatory responses. This study aimed to investigate the role of HNF4α in the development of fulminant hepatic failure (FHF) induced by lipopolysaccharide/D-galactosamine (LPS/D-GalN). METHODS: The FHF model was induced by simultaneous intraperitoneal injection of LPS/D-GalN in mice. Three days prior to LPS/D-GalN administration, HNF4α short-hairpin interfering RNA expression plasmid or physiological saline was injected via the tail vein with a hydrodynamics-based procedure. The degree of hepatic damage and cumulative survival rate were subsequently assessed. RESULTS: The expression of HNF4α was increased in the early stage after LPS/D-GalN administration. Inhibiting the expression of HNF4α reduced serum levels of alanine aminotransferase and aspartate aminotransferase, alleviated histological injury, and improved the survival of mice with FHF. In addition, both serum and hepatic tumor necrosis factor alpha expression were suppressed when HNF4α expression was inhibited in mice with FHF. CONCLUSION: Inhibiting HNF4α expression protects mice from FHF induced by LPS/D-GalN, but the exact mechanism behind this needs further investigation.

Protective Activity of <i>Markhamia tomentosa</i>(Benth.) K. Schum. (Bignoniaceae) Methanol Leaves Extract against D-Galactosamine/Lipopolysaccharide-Induced Acute Liver Injury in Mice

Markhamia tomentosa (Benth.) K. Schum. (Mt) is a Cameroonian medicinal plant, traditionally used to treat painful and inflammatory illness. This study aimed to examine the effects of methanol leaves extract (MLE) of Mt in D-galactosamine (D-GaIN)/lipopolysaccharide (LPS)-induced liver injury. The MLE (100 and 200 mg/kg), Ascorbic acid (10 mg/kg) and distilled water were administered 12 h and 1 h before intraperitoneal injection of D-GaIN (10 mg/mouse)/LPS (0.1 μg/g). Animals were sacrificed 6 h after D-GalN/LPS challenge. Liver injury was assessed biochemically by determination of aspartate aminotransferase (ASAT), alanine aminotransferase (ALAT), alkaline phosphatase (ALP), superoxide dismutase (SOD) and catalase activities. Malondialdehyde (MDA), reduced glutathione (GSH), nitrites, total protein and bilirubin levels were explored. Histopathological examination of liver tissue was also performed. Liver enzymes (ALAT, ASAT, ALP) activity, nitrites, MDA and bilirubin levels were increased, while protein level, SOD and catalase activities were significantly reduced by D-GalN/LPS administration. MLE (100 or 200 mg/kg) protected mice against D-GalN/LPS-induced death. In addition, the plant extract significantly reduced ALAT and ALP activity, exhibiting 23.00% and 62.20% protection, respectively. SOD activity and total protein were significantly (p < 0.05) increased by the plant extract. Total bilirubin and MDA levels were reduced (p < 0.01) by 37.75% and 62.79%, respectively in animal treated with MLE. Histological analysis of liver sections showed that MLE (100 or 200 mg/kg) protected mice against D-GaIN/LPS-induced liver injury. The obtained results showed that MLE of Mt may possess hepatoprotective effects. Protection afforded by MLE against D-GalN/LPS-induced fulminant liver injury may result from reduction of oxidative stress.

Protective Effects of Self-made Compound Taoren Danshen Decoction on Rats with Acute Liver Injury

[Objectives] To study the protective effects of self-made Compound Taoren Danshen Decoction( CTDD) on acute liver injury induced by D-galactosamine in rats,and to explore its mechanism. [Methods]An acute liver injury model was established by intraperitoneal injection of 500 mg/kg of D-galactosamine. The ALT,AST,MDA,SOD and GSH-Px in serum,as well as serum TNF-α,IL-1β and IL-6 levels were measured,and liver tissue lesions were observed under microscope. [Results] CTDD can significantly reduce ALT,AST and MDA levels,increase SOD,GSH-Px activity,and significantly reduce serum TNF-α,IL-1β and IL-6 levels,and improve liver tissue lesions.[Conclusions]CTDD has a protective effect on D-galactosamine-induced acute liver injury in rats,and its mechanism may be related to inhibition of oxidative stress and inflammatory reaction.

Protective Effect of the Zhuang Medicine Cryptolepis buchananii Roem. et Schult. on D-GalN-induced Liver Injury in Mice

[Objectives] This study was conducted to study the protective effect of the Zhuang medicine Cryptolepis buchananii Roem. et Schult. on D-galactosamine(D-GalN)-induced liver injury in mice. [Methods] A mouse model of chemical liver injury was established by D-GalN, and the mice with D-GalN-induced liver injury were randomly divided into the blank group, model group, positive control group and drug-administered groups. The levels of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) in the serum of the mice in each group and the levels of superoxide dismutase(SOD) and malondialdehyde(MDA) in the liver tissue of the mice were determined, and the liver pathological changes were observed. [Results] The extracts of the Zhuang medicinal herb C. buchananii could reduce the serum ALT and AST levels(P<0.05), increase the SOD content in the liver tissue of mice(P<0.05), and decrease the MDA level(P<0.05). [Conclusions] The Zhuang medicinal herb C. buchananii has a good protective effect on D-GalN-induced liver injury in mice.

Establishment of a new acute-on-chronic liver failure model

To establish an animal model of acute-on-chronic liver failure(ACLF) that would replicate the pathological process of ACLF in humans, rats were administered porcine serum(PS) for 11 weeks. Liver fibrosis was determined by pathological and biochemical assessments. The animals then were injected with D-galactosamine(D-gal) and lipopolysaccharide(LPS). The survival times of animals with cirrhosis and ACLF were determined over 48 h. Other animals were killed at 0, 4, 8 and 12 h after administration of D-gal/LPS. Liver injury was assessed by histopathological analysis and biochemical indices, and apoptosis was detected by Western blot and TUNEL analysis. After PS administration for 11 weeks the serum levels of hyaluronic acid and N-procollagen type III peptide increased significantly, and serious fibrosis and cirrhosis was observed at weeks 10 and 11. Cirrhotic rats were injected with D-gal/LPS to induced ACLF; the rate of mortality over 48 h was 80%. ALT and AST levels increased markedly at 4 h, but decreased significantly at8 and 12 h post-treatment. The total bilirubin, direct bilirubin, and total bile acids levels increased markedly at 8 and 12 h. Clotting times, TNF-α and IL-6 levels increased significantly, except for 12 h post-treatment.Apoptosis, inflammation and necrosis were elevated as determined by hematoxylin-eosin staining and TUNEL assays. BCL-2 levels decreased significantly, While BAX levels increased significantly.Cytochrome c expression peaked at 8 h postD-gal/LPS treatment. In conclusion, an ACLF model induced by PS and D-gal/LPS was established and the underlying mechanisms of ACLF development were explored.

Adiponectin receptor agonist AdipoRon relieves endotoxin-induced acute hepatitis in mice

Background:Adiponectin is the most abundant adipokines that plays critical roles in the maintenance of energy homeostasis as well as inflammation regulation. The half-life of adiponectin is very short and the small-molecule adiponectin receptor agonist has been synthesized recently. In the present study, the potential roles of AdipoRon, an adiponectin receptor agonist, in a mouse model of lipopolysaccharide (LPS)/D-galactosamine (D-Gal)-induced acute hepatitis was explored.Methods:BALB/c mice (n = 144, male) were divided into three sets. In set 1, 32 mice were randomized into four groups: the control group, the AdipoRon group, the LPS/D-Gal group, and the AdipoRon + LPS/D-Gal group. The mice in set 1 were sacrificed after LPS/D-Gal treatment, and the plasma samples were collected for detection of tumor necrosis factor-alpha (TNF-α). In set 2, the 32 mice were also divided into four groups similar to that of set 1. The mice were sacrificed 6 h after LPS/D-Gal injection and plasma samples and liver were collected. In set 3, 80 mice (divided into four groups, n = 20) were used for survival observation. The survival rate, plasma aminotransferases, histopathological damage were measured and compared between these four groups.Results:AdipoRon suppressed the elevation of plasma aminotransferases (from 2106.3 ± 781.9 to 286.8 ± 133.1 U/L for alanine aminotransferase, P < 0.01;from 566.5 ± 243.4 to 180.1 ± 153.3 U/L for aspartate aminotransferase, P < 0.01), attenuated histopathological damage and improved the survival rate (from 10% to 60%) in mice with LPS/D-Gal-induced acute hepatitis. Additionally, AdipoRon down-regulated the production of TNF-α (from 328.6 ± 121.2 to 213.4 ± 52.2 pg/mL, P < 0.01), inhibited the activation of caspase-3 (from 2.04-fold to 1.34-fold of the control), caspase-8 (from 2.03-fold to 1.31-fold of the control), and caspase-9 (from 2.14-fold to 1.43-fold of the control), and decreased the level of cleaved caspase-3 (0.28-fold to that of the LPS/D-Gal group). The number of terminal deoxynucleotidyl transferase-mediated nucleotide nick-end labeling-positive apoptotic hepatocytes in LPS/D-Gal-exposed mice also reduced.Conclusions:These data indicated that LPS/D-Gal-induced acute hepatitis was effectively attenuated by the adiponectin receptor agonist AdipoRon, implying that AdipoRon might become a new reagent for treatment of acute hepatitis.

Effect of Magnesium Lithosperarnate B on D－galactosamine Induced Liver Injury in Rats

Treated the experimental acute liver injury rat induced by D-galactosamine with magnesiumIithosperamate-B could significantly lower the serum activity of alanine transaminase and aspartate amino-transferase, relieve the necrosis of hepatocytes and elevate the levels of products of arachidonic acid (6-ke-to-prostaglandin 1 , prostaglandin D2 and the total prostaglandin) in liver. These results suggested that mag-nesium lithosperamate B was one of the rnain components of Salvia miltiorrzae in treating liver injury, the ac-tion on elevating the total prostaglandin in liver might be its mechanism.