Dimethylnitrosamine Research Articles

Huangqi decoction alleviates dimethylnitrosamine-induced liver fibrosis: An analysis of bile acids metabolic mechanism

Ethnopharmacological relevanceHuangqi Decoction (HQD), a classical traditional Chinese medicine (TCM) formula, is used to treating liver injury in China. The aim of the study is to investigate mechanisms of HQD against dimethylnitrosamine (DMN)-induced liver fibrosis underlying metabolic profiles of bile acids. Materials and methodsDMN-induced liver fibrosis rats were administrated HQD and its compounds, astragalosides (AS), glycyrrhizic acid (GA) and their combination. The anti-fibrosis effects were evaluated and targeted metabolomics by UPLC-MS was used to examine whether HQD had an influence on bile acid metabolism. The levels of mRNAs associated with bile acid metabolism were expressed by RT-PCR. Chenodeoxycholic acid (CDCA)-induced hepatic stellate cells (HSCs) proliferation and activation were examined using MTS assay and Western blot. ResultsHistopathological changes and serum liver function in HQD group had significant improvements (P<0.01). Concentrations of free bile acids and taurine conjugates were significantly increased in DMN group (P<0.05). HQD and its compounds restored the increased bile acids to normal levels, and HQD was more effected on parts of bile acids. Furthermore, the levels of mRNAs related bile acid synthesis and reabsorption such as CYP7A1, CYP8B1, CYP27A1, OATP2, OATP3, OATP4 and NTCP were significantly down-regulated in DMN group (P<0.05), mRNAs related excretion such as MRP3 and BESP were up-regulated (P<0.01), and CYP7A1, CYP8B1, OATP3, OATP4, NTCP and MRP3 restored to normal levels by HQD treatment. Moreover, CDCA-induced HSCs proliferation and activation were weaken by HQD (P<0.05) with down-regulated α-SMA, TGF-β1, p-Smad2 and p-Smad3 expressions. ConclusionsHQD alleviated DMN-induced liver fibrosis with a better effect than its compounds, which may be involved in the regulation of bile acid metabolism enzyme. Moreover, HQD may inhibit CDCA-induced HSCs proliferation and activation.

Dimethylnitrosamine (DMN) pre-treated rats and protective effect of Vernonia amygdalina post-treatment on liver function

Background: Liver disease and toxicity is common, especially with many drug treatments. The effect of Vernonia amygdalina ethanolic leaf extract post-treatment on rats pre-treated with dimethylnitrosamine (DMN) was evaluated.Methods: Male wistar rats were orally administered DMN (single dose of 25 mg/kg) on first day and thereafter post-treated with 100 and 200mg/kg ethanolic leaf extract of Vernonia amygdalina for seven days. Analysis of serum concentrations of albumin, total protein, total bilirubin and glucose were carried out.Results: Administration of DMN alone to rats significantly increased glucose and total bilirubin concentration (P &lt; 0.05) in the serum while it significantly reduced (P&lt;0.05) serum total protein and albumin concentrations when compared with controls. However, post-treatment of DMN administered rats with 100 and 200 mg/kg ethanolic extract leaf of Vernonia amygdalina significantly (P&lt;0.05) reversed these changes in a concentration dependent manner.Conclusion: In conclusion ethanolic leaf extract of Vernonia amygdalina have a potent hypoglycaemic and hepatoprotective action against dimethylnitrosamine-induced liver damage in rats may be due to its antioxidant properties.

Chinese medicine CGA formula ameliorates DMN-induced liver fibrosis in rats via inhibiting MMP2/9, TIMP1/2 and the TGF-β/Smad signaling pathways.

Chinese medicine CGA formula consists of polysaccharide from Cordyceps sinensis mycelia (CS-PS), gypenosides and amygdalin, which is derived from Fuzheng Huayu (FZHY) capsule for treating liver fibrosis. In this study we attempted to confirm the therapeutic effects of CGA formula in dimethylnitrosamine (DMN)-induced liver fibrosis in rats, and to identify the mechanisms of anti-fibrotic actions. Rats were injected with DMN (10 mg·kg(-1)·d(-1), ip) for 3 consecutive days per week over a 4-week period. The rats then were orally administered with CGA formula (CS-PS 60 mg·kg(-1)·d(-1), gypenosides 50 mg·kg(-1)·d(-1) and amygdalin 80 mg·kg(-1)·d(-1)) daily in the next 2 weeks. CS-PS, gypenosides or amygdalin alone were administered as individual component controls, whereas colchicine and FZHY were used as positive controls. Serum biomarkers were measured. Hepatic injury, collagen deposition and stellate cell activation were examined. The MMP activities, expression of TIMP protein and proteins involved in the TGF-β1/Smad signaling pathways in liver tissues were assayed. In DMN-treated rats, administration of CGA formula significantly decreased serum ALT, AST and total bilirubin and hepatic hydroxyproline levels, increased serum albumin level, and attenuated liver fibrosis as shown by histological examination. Furthermore, these effects were comparable to those caused by administration of FZHY, and superior to those caused by administration of colchicine or the individual components of CGA formula. Moreover, administration of CGA formula significantly decreased the protein levels of α-SMA, TGF-β1, TGF-β1 receptor (TβR-I), p-TβR-I, p-TβR-II, p-Smad2, p-Smad3, TIMP1 and TIMP2, as well as MMP2 and MMP9 activities in liver tissues of DMN-treated rats. Chinese medicine CGA formula ameliorates DMN-induced liver fibrosis in rats, and this effect was likely associated with the down-regulation of MMP2/9 activities, TIMP1/2 protein expression and the TGF-β1/Smad signaling pathways in the liver.

Structural transition of kidney cystatin in dimethylnitrosamine-induced renal cancer in rats: identification as a novel biomarker for kidney cancer and prognosis

In our study, renal cancer is induced in rats making use of dimethylnitrosamine (DMN). G1 – Group 1 were control rats and G2 – Group 2 rats were given a single intra-peritoneal injection of DMN of 50 mg/kg body weight resulting in 100% incidences of renal tumors after 12 months. SEM and histopathology confirmed the presence of renal cancer in the DMN-treated rats. Making use of ammonium sulfate precipitation and gel filtration chromatography on Sephacryl S-100HR column, a thiol protease inhibitor was isolated from kidney of control rats known as Rat kidney Cystatin (RKC) as well as from kidney of cancerous rat called as Cancerous Rat Kidney Cystatin (CRKC). Both these inhibitors were characterized, and interestingly, it was found that CRKC showed greater anti-papain activity and also it was stable in a broad pH and temperature range thus implying that CRKC is more stable as compared to RKC. UV and fluorescence spectroscopy point out in structural difference between RKC and CRKC which was further confirmed by Circular dichroism (CD) and FTIR spectroscopy. Our study clearly showed that kidney cystatin is structurally modified in the case of renal cancer and performs its role in a more efficacious manner.

Localization of Xenobiotic Transporter OCTN1/SLC22A4 in Hepatic Stellate Cells and Its Protective Role in Liver Fibrosis

Xenobiotic transporters play key roles in disposition of certain therapeutic agents, although limited information is available on their roles other than pharmacokinetic issues. Here, suppressive effect of multispecific organic cation transporter OCTN1/SLC22A4 on liver fibrosis was proposed in liver injury models. After injection of hepatotoxins such as dimethylnitrosamine (DMN) or concanavalin A, hepatic fibrosis, and oxidative stress, evaluated in terms of Sirius red and 4-hydroxy-2-nonenal staining, respectively, were more severe in liver of octn1/slc22a4 gene knockout (octn1−/−) mice than that in wild-type mice. DMN treatment markedly increased α-smooth muscle actin and F4/80, markers of activated stellate and Kupffer cells, respectively, in liver of octn1−/−, but had less effect in wild-type mice. Thus, octn1/slc22a4 gene deletion results in more severe hepatic fibrosis, oxidative stress, and inflammation. DMN-treated wild-type mice showed increased Octn1 staining and hepatic concentration of its food-derived antioxidant ergothioneine (ERGO). The upregulated Octn1 was co-localized with α-smooth muscle actin. Functional expression of Octn1 was demonstrated in activated human hepatic stellate cell lines, LI90 and LX-2. Provision of ERGO-rich feed ameliorated DMN-induced liver fibrosis and oxidative stress. Overall, Octn1 is upregulated in activated stellate cells, resulting in increased delivery of its substrate antioxidant ERGO and a protective effect against liver fibrosis.

Dimethylnitrosamine (DMN) exposed rats: Vernonia amygdalina pre-treatment enhances immunity, hepatic and renal function

&lt;p&gt;&lt;strong&gt;Background:&lt;/strong&gt; The occurrence of dietary and environmental chemicals such as dimethylnitrosamine (DMN) in drinks and foods including fish and meat as well as fresh supermarket products is well established. This study evaluated protective effect of ethanolic leaf extract of &lt;em&gt;Vernonia amygdalina&lt;/em&gt; (VAE) on liver synthetic molecules, kidney function and hematological parameters in acute dimethylnitrosamine (DMN)-induced hepatic toxicity in wistar male rats.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods:&lt;/strong&gt; Experimental rats divided into four groups of six rats each were used. The first group was untreated and served as control. The second group was orally administered VAE (400 mg/kg) only for seven days. The third group was pre-treated with VAE (400mg/kg) for 7 days and administered 20mg/kg DMN 24hrs after VAE pre-treatment. Rats of the fourth group were given 20mg/kg DMN alone same time with that of group 3. All rats were sacrificed 48hrs after DMN administration.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results:&lt;/strong&gt; In rats administered 20mg/kg DMN, VAE pre-treatment at 400 mg/kg significantly increased total protein, albumin,White blood cell (WBC), Red blood cell (RBC), Hemoglobin (Hb), packed cell volume (PCV) and Platelets while it significantly decreased total bilirubin, urea and creatinine compared to DMN-alone administered rats.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusion:&lt;/strong&gt; This study suggest that VAE pre-treatment exert its ameliorative effect against DMN-induced hematological and biochemical alterations possibly by preventing the decline of antioxidant defense system and could be prescribed as adjunct to dietary therapy.&lt;/p&gt;

Dimethylnitrosamine (DMN)-induced fibrotic rats: effect of Vernonia amygdalina on extracellular matrix and Hepatic/lysosomal integrity

&lt;p&gt;&lt;strong&gt;Background:&lt;/strong&gt; Hepatic fibrosis is the accumulation of extracellular matrix protein, or scar, in response to acute or chronic liver injury. This study investigated the effect of ethanolic leaf extract of &lt;em&gt;Vernonia amygdalina&lt;/em&gt; on extracellular matrix protein: hyaluronic acid (HA), liver synthetic molecules: total protein (TP), albumin (ALB), total bilirubin (TB) as well as lysosomal membrane stability: acid phosphatase (ACP) in dimethylnitrosamine (DMN)-induced fibrotic rats.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods:&lt;/strong&gt; Wistar albino male rats were intraperitoneally injected with 10 mg/kg DMN on first three days a week for two weeks. Ethanolic leaf extracts of &lt;em&gt;Vernonia amygdalina&lt;/em&gt; (200 mg /kg) was administered simultaneously by oral gavage daily for two weeks. All rats were sacrificed after 24 hours of last administration by cardiac puncture, and blood collected from the ocular vein. Analysis of serum ACP and LDH activities with those of the concentrations of HA, ALB, TP and TB were carried out.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results:&lt;/strong&gt; Administration of DMN to rats significantly increased HA and TB concentration and the activities of ACP and LDH (p&amp;lt;0.05) in the serum while it significantly reduced (p&amp;lt;0.05) serum TP and ALB concentrations when compared with controls. However, simultaneous administration of ethanolic leaf extracts of &lt;em&gt;Vernonia amygdalina&lt;/em&gt; with DMN significantly (p&amp;lt;0.05) reversed these changes.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusion:&lt;/strong&gt; This study shows that &lt;em&gt;Vernonia amygdalina&lt;/em&gt; possesses hepatoprotective, lysosomal membrane stabilizing and anti-fibrotic properties may be due to its antioxidant and phytochemical constituents.&lt;/p&gt;

Annona muricata prevent hepatic fibrosis by enhancing lysosomal membrane stability and suppressing extracellular matrix protein accumulation

Background: Hepatic fibrosis is a pre-pathologic state of cirrhosis, which plays a pivotal role in the pathogenesis of hepatocellular carcinoma (HCC). This study was carried out to evaluate the hepatoprotective effect of Annona muricata leaves on extracellular matrix (ECM) accumulation, lysosomal membrane integrity and liver damage in dimethylnitrosamine (DMN)-induced fibrotic rats.Methods: Healthy adult male wistar rats were divided into four groups. Group I rats received normal saline and served as control, Group II rats were administered 200mg/kg Annona muricata, Group III received 200mg/kg Annona muricata orally simultaneously with 10mg/kg DMN, Group IV received 10 mg/kg DMN without extract. DMN administered intraperitoneally was on first three days of each week for two weeks while Annona muricata was administered for 14 consecutive days. 24 hours after last administration, all animals were sacrificed and blood collected for serum analysis. Biochemical parameters such as lactate dehydrogenase (LDH), total bilirubin (TB), total protein (TP), and albumin (ALB) were estimated to assess liver function and synthetic ability while hyaluronic acid (HA) was determined to assess extracellular matrix (ECM) function. In addition, acid phosphatase (ACP) was estimated to assess lysosomal membrane stability/fragility.Results: Liver damage, decreased synthetic ability, lysosomal membrane fragility and altered ECM function were evident by an increase in the levels of LDH, TB, ACP and HA as well as a decrease in level of TP and ALB in sera of rats administered DMN compared to normal rats. However, simultaneous treatment with Annona muricata leaf extract significantly (p&lt;0.05) reversed alterations in the above indices.Conclusion: This study suggests that Annona muricata leaf functions as a potent fibrosuppressant by suppressing ECM accumulation, enhancing lysosomal membrane stability and liver synthetic ability.

Serum Metabolomic Characterization of Liver Fibrosis in Rats and Anti-Fibrotic Effects of Yin-Chen-Hao-Tang.

Yin-Chen-Hao-Tang (YCHT) is a famous Chinese medicine formula which has long been used in clinical practice for treating various liver diseases, such as liver fibrosis. However, to date, the mechanism for its anti-fibrotic effects remains unclear. In this paper, an ultra-performance liquid chromatography-time-of-flight mass spectrometry (UPLC-TOF-MS)-based metabolomic study was performed to characterize dimethylnitrosamine (DMN)-induced liver fibrosis in rats and evaluate the therapeutic effects of YCHT. Partial least squares-discriminant analysis (PLS-DA) showed that the model group was well separated from the control group, whereas the YCHT-treated group exhibited a tendency to restore to the controls. Seven significantly changed fibrosis-related metabolites, including unsaturated fatty acids and lysophosphatidylcholines (Lyso-PCs), were identified. Moreover, statistical analysis demonstrated that YCHT treatment could reverse the levels of most metabolites close to the normal levels. These results, along with histological and biochemical examinations, indicate that YCHT has anti-fibrotic effects, which may be due to the suppression of oxidative stress and resulting lipid peroxidation involved in hepatic fibrogenesis. This study offers new opportunities to improve our understanding of liver fibrosis and the anti-fibrotic mechanisms of YCHT.

Effect of Annona muricata pre-treatment on liver synthetic ability, kidney function and hematological parameters in dimethylnitrosamine (DMN)-administered rats

Background:Dimethylnitrosamine (DMN) is a potent hepatotoxin, carcinogen and mutagen which exerts carcinogenic effects and induces hepatic necrosis in experimental animals through metabolic activation by CYP2E1. This study thus evaluated efficacies of Annona muricata pre-treatment on acute dimethylnitrosamine (DMN)-induced anemia and biochemical alterations in male albino rats.Methods:Four (4) groups of six (6) rats each were used for the study. Group 1 served as control and was untreated, group 2 and 3 were pre-treated with 400mg/kg Annona muricata ethanol leaf extract for one week while group 3 and 4 each received single dose of 20mg/kg DMN (orally) after one week. The rats were sacrificed 48hrs after DMN administration.Results:In rats administered 20mg/kg DMN, toxicty was clearly shown by decreased activities of serum liver synthetic molecules namely total protein, albumin, increased total bilirubin, increased kidney parameters such as creatinine and urea as well as decreased body weight gain. The toxic effect of DMN was also indicated by significantly decreased levels of red blood cells (RBC), hemoglobin (Hb), packed cell volume (PCV), white blood cells (WBC) and platelet count (Plt count). However, in rats pre-treated with 400mg/kg Annona muricata prior to DMN administration, there were significant reversals in the activities of serum liver synthetic molecules, kidney profiles, and hematological parameters when compared to DMN-alone administered rats.Conclusion:Annona muricata pre-treatment exhibited in vivo protective and anti-anemic effects against DMN-induced injury.

Improvement of Gene Expression Studies in the Dimethylnitrosamine Induced Liver Fibrosis Model in the Rat Using Selected Reference Genes for Quantitative Real Time-PCR Analysis

Background: Liver fibrosis is a reaction to chronic liver injury characterized by excessive accumulation of collagen. Due to their importance as biomarkers, the changes in gene expression in the liver during the development of fibrosis and its subsequent outcomes of cirrhosis, neoplasia or resolution are intensely studied. Quantitative realtime PCR (qPCR) with its ability to detect and measure minute amounts of nucleic acids have been increasingly used in these studies. In qPCR, the quantitation of mRNA is relative and the accuracy of results dependent on the reference genes used for standardization. However, many genes studied are normalized against single reference genes, usually housekeeping genes, without adequate justification. Methods: For the dimethylnitrosamine (DMN) induced liver fibrosis rat model, we tested 8 commonly used candidate genes (Actb, Alb, Sdha, B2m, Rn18s, Hprt1, Ppia and Gapdh) to determine their suitability as reference genes. qPCR results were analysed using four commonly used programs; NormFinder, GeNorm, Comparative ΔCt methods and BestKeeper. Result: It was determined that Gapdh and B2m were the most stable genes in normal liver. However, in DMN treated livers, Gapdh and Ppia were the most stably expressed reference genes. We validated these reference genes by using them to normalize the expression of four genes; Tgfb 1, Col1a1, Col3a1 and Tnf known to be highly expressed in liver fibrosis. Conclusion: Gapdh and Ppia are the most suitable reference genes for the normalization of qPCR data in gene expression studies of the liver in the DMN induced liver fibrosis model in the rat. We advise against the use of Actb in this experimental setting because of its low expression stability.

Ethyl Acetate Fraction of Amomum xanthioides Exerts Antihepatofibrotic Actions via the Regulation of Fibrogenic Cytokines in a Dimethylnitrosamine-Induced Rat Model.

Amomum xanthioides has been traditionally used to treat diverse digestive system disorders in the Asian countries. We investigated antihepatofibrotic effects of ethyl acetate fraction of Amomum xanthioides (EFAX). Liver fibrosis is induced by dimethylnitrosamine (DMN) injection (intraperitoneally, 10 mg/kg of DMN for 4 weeks to Sprague-Dawley rats). EFAX (25 or 50 mg/kg), silymarin (50 mg/kg), or distilled water was orally administered every day. The DMN injection drastically altered body and organ mass, serum biochemistry, and platelet count, while EFAX treatment significantly attenuated this alteration. Severe liver fibrosis is determined by trichrome staining and measurement of hydroxyproline contents. EFAX treatment significantly attenuated these symptoms as well as the increase in oxidative by-products of lipid and protein metabolism in liver tissues. DMN induced a dramatic activation of hepatic stellate cells and increases in the levels of protein and gene expression of transforming growth factor-beta (TGF-β), platelet derived growth factor-beta (PDGF-β), and connective tissue growth factor (CTGF). Immunohistochemical analyses revealed increases in the levels of protein and gene expression of α-smooth muscle actin. These alterations were significantly normalized by EFAX treatment. Our findings demonstrate the potent antihepatofibrotic properties of EFAX via modulation of fibrogenic cytokines, especially TGF-β in the liver fibrosis rat model.

Cirrhosis Model That Tracks Umbilical Cord Mesenchymal Cell Transplantation Using Liver Serum Parameters: A Pilot Study

AIMS: This study aims to evaluate a preclinical model of hepatic failure to propose an effective scheme to follow progression of mesenchymal cell treatments using serum parameters. METHODS: Male Wistar rats (n = 53) received dimethylnitrosamine (DMN) or PBS (control) intraperitoneally. We evaluated liver macroscopy, histology, serum albumin (ALB), total bilirubin (TB), alanine (ALT) and aspartate (AST) aminotransferase, alkaline phosphatase (ALP), gamma globulin, and hepatic gene expression of ALB and selected regeneration markers. RESULTS: After defining the effective DMN dose, 1 × 107 umbilical cord mesenchymal stem cells (UC-MSC) were injected into the tail vein 1 week after treatment and liver function re-assessed. DMN 10 µg/g delivered 3 days/week for 4 weeks altered liver macroscopy and histology. DMN treatment also increased ALP, ALT and TB, and significantly reduced gene expression of regeneration factors, although ALB and hepatic growth factor mRNA were not altered. Upon UC-MSC treatment, a greater percentage of rats showed reduction of AST levels (75% UC-MSC vs 0% PBS). CONCLUSIONS: Selected serum parameters can predict liver function in a rodent model, but each animal must be its own control.

Umbilical cord-derived mesenchymal stem cells alleviate liver fibrosis in rats.

To evaluate the efficacy of umbilical cord-derived mesenchymal stem cells (UC-MSCs) transplantation in the treatment of liver fibrosis. Cultured human UC-MSCs were isolated and transfused into rats with liver fibrosis induced by dimethylnitrosamine (DMN). The effects of UC-MSCs transfusion on liver fibrosis were then evaluated by histopathology; serum interleukin (IL)-4 and IL-10 levels were also measured. Furthermore, Kupffer cells (KCs) in fibrotic livers were isolated and cultured to analyze their phenotype. Moreover, UC-MSCs were co-cultured with KCs in vitro to assess the effects of UC-MSCs on KCs' phenotype, and IL-4 and IL-10 levels were measured in cell culture supernatants. Finally, UC-MSCs and KCs were cultured in the presence of IL-4 antibodies to block the effects of this cytokine, followed by phenotypical analysis of KCs. UC-MSCs transfused into rats were recruited by the injured liver and alleviated liver fibrosis, increasing serum IL-4 and IL-10 levels. Interestingly, UC-MSCs promoted mobilization of KCs not only in fibrotic livers, but also in vitro. Co-culture of UC-MSCs with KCs resulted in increased production of IL-4 and IL-10. The addition of IL-4 antibodies into the co-culture system resulted in decreased KC mobilization. UC-MSCs could increase IL-4 and promote mobilization of KCs both in vitro and in vivo, subsequently alleviating the liver fibrosis induced by DMN.

Hepatoprotective effects of Dicliptera chinensis polysaccharides on dimethylnitrosamine-induced hepatic fibrosis rats and its underlying mechanism

Ethnopharmacological relevanceDicliptera chinensis is a traditional herbal medicine used anciently in China for hepatopathy treatment, especially in south areas. Our several studies have demonstrated that dicliptera chinensis polysaccharides (DCP), which has a markedly protective effects on chemistry-induced models of acute liver injury in rats. In this study, we further investigated the potentially hepatoprotective effect of dicliptera chinensis polysaccharides (DCP) on hepatic fibrosis (HF) rats induced by dimethylnitrosamine (DMN). Material and methodsThe 96 rats were randomly divided into six groups (n=16, per group), the normal control group intragastrically administrated normal saline, model control group intraperitoneally injected with 0.5% DMN solution at 1.6mL per kg (three times a week); colchicine intragastrically administrated group (0.2mgkg−1d−1)+DMN-treated rats; DCP intragastrically administrated groups (100mgkg−1d−1, 200mgkg−1d−1, 300mgkg−1d−1)+DMN-treated rats. At the end of 8 weeks, all rats were sacrificed. ResultsPathological examination showed that high and medium doses of DCP presented remarkable effect in ameliorating hepatic fibrosis, alleviate the inflammation, necrosis and reduced collagen deposits. DCP effectively improved the liver function, as revealed in being lowered sero-enzyme levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL) while increased albumin (ALB), and being reduced sero-concentrations of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6) in the HF rats. Additionally, the contents of hyaluronic acid (HA), collagen type Ⅳ (Ⅳ-C), type III precollagen (PCIII) and laminin (LN) in the hepatic tissue of HF rats were markedly decreased, whereas the expressions of transforming growth factor-β l (TGF-β l), collagen type I (Col- I), metal protease-1 (TIMP-1), nuclear factor-kappa B (NF-κB) expression in the hepatic tissue were notably down-regulated. ConclusionDCP exerts effectively antagonistic activity on DMN-caused hepatotoxicity in HF rats, which the anti-fibrotic mechanisms are associated with regulating functionally serous enzymes, improving metabolic function and inhibiting inflammatory reaction in liver tissue.

Hepatoprotective effects of zingerone on carbon tetrachloride- and dimethylnitrosamine-induced liver injuries in rats.

In this study, we investigated the hepatoprotective and anti-fibrotic effects of zingerone, one of the active components of ginger, against carbon tetrachloride (CCl4)- and dimethylnitrosamine (DMN)-induced liver injuries in rats, respectively. Oral administration of zingerone (10mg/kg) reduced CCl4-induced abnormalities in liver histology, serum alanine aminotransferase and aspartate aminotransferase levels, and liver malondialdehyde levels. Zingerone treatment attenuated CCl4-induced increases in inflammatory mediators, including tumor necrosis factor-α, interleukin-1β, cyclooxygenase-2, and inducible nitric oxide synthase mRNA levels. Western blot analysis showed that zingerone suppressed activation of nuclear factor-kappa B (NF-κB) p65 and phosphorylation of extracellular signal-regulated kinase, c-Jun NH2-terminal kinase, and p38 mitogen-activated protein kinases (MAPKs). Liver fibrosis induced by DMN (10mg/kg, intraperitoneally) was ameliorated by administration of zingerone (10 and 20mg/kg, orally). Zingerone treatment reduced DMN-induced elevation of hydroxyproline content and hepatic stellate cell activation. In conclusion, zingerone showed antioxidative and anti-inflammatory effects in CCl4-intoxicated rats by inhibiting oxidative stress and NF-κB activation via blockade of the activation of upstream MAPKs. Moreover, zingerone had hepatoprotective and anti-fibrotic effects against DMN-induced liver injury suggesting its usefulness in the prevention of liver inflammation and the development of hepatic fibrosis.

Actions of Huangqi decoction against rat liver fibrosis: a gene expression profiling analysis.

BackgroundHuangqi decoction (HQD) is used for liver fibrosis and cirrhosis treatment in Chinese medicine. This study aims to investigate the pharmacological actions of HQD against liver fibrosis in rats by high-throughput gene expression profiling, network analysis and real-time qRT-PCR.MethodsWe analyzed the profiles of differentially expressed genes (DEGs) in dimethylnitrosamine (DMN)-induced liver fibrosis in rat. The liver tissue samples of control group (n = 3), model group (n = 3) and HQD group (n = 3) were examined by microarrays. Pathways were analyzed by KEGG. Pathway-gene and protein–protein interaction (PPI) networks were constructed with Cytoscape software. The expression of candidate genes was verified by qRT-PCR. P values less than 0.05 indicated statistical significance.ResultsCollagen deposition and hydroxyproline (Hyp) content were decreased in the HQD group compared with the model group (P < 0.001), while that of Hyp in the model group were increased compared with the control group (P < 0.001). In comparison with the model group, 1085 DEGs (all P < 0.05, |fold change| >1.5) and 52 pathways in the HQD group were identified. TGF-beta, ECM-receptor interaction, and the cell adhesion molecules pathways were significantly recovered by HQD (P < 0.001). A pathway-gene network was constructed, including 303 DEGs and 52 pathways, and 514 nodes and 2602 edges, among 142 genes with node degrees greater than 10. The expressions of PDGFra, PDGFrb, PDGFb, PDGFd, COL1A1, COL1A2, COL5A2, and THBS1 were significantly down-regulated by HQD (P < 0.001).ConclusionHQD down-regulated the expressions of PDGFra, PDGFrb, PDGFb, PDGFd, COL1A1, COL1A2, COL5A2 and THBS1, and TGF-β and PDGF signaling pathways in the DMN-induced liver fibrosis in rats.Electronic supplementary materialThe online version of this article (doi:10.1186/s13020-015-0066-5) contains supplementary material, which is available to authorized users.

Fibroblast growth factor 21 attenuates hepatic fibrogenesis through TGF-β/smad2/3 and NF-κB signaling pathways

Fibroblast growth factor 21 (FGF-21) is a secreted protein, which has anti-diabetic and lipocaic effects, but its ability to protect against hepatic fibrosis has not been studied. In this study, we investigated the ability of FGF-21 to attenuate dimethylnitrosamine (DMN)-induced hepatic fibrogenesis in mice and the mechanism of its action. Hepatic fibrosis was induced by injection of DMN, FGF-21 was administered to the mice once daily in association with DMN injection till the end of the experiment. Histopathological examination, tissue 4-hydroxyproline content and expressions of smooth muscle α-actin (α-SMA) and collagen I were measured to assess hepatic fibrosis. Ethanol/PDGF-BB-activated hepatic stellate cells (HSCs) were used to understand the mechanisms of FGF-21 inhibited hepatic fibrogenesis. Results showed that FGF-21 treatment attenuated hepatic fibrogenesis and was associated with a significant decrease in intrahepatic fibrogenesis, 4-hydroxyproline accumulation, α-SMA expression and collagen I deposition. FGF-21 treatment inhibited the activation of HSCs via down-regulating the expression of TGF-β, NF-κB nuclear translocation, phosphorylation levels of smad2/3 and IκBα. Besides, FGF-21 treatment caused activated HSC apoptosis with increasing expression of Caspase-3, and decreased the ratio of Bcl-2 to Bax. In conclusion, FGF-21 attenuates hepatic fibrogenesis and inhibits the activation of HSC warranting the use of FGF-21 as a potential therapeutic agent in the treatment of hepatic fibrosis.

Effect of Opisthorchis felineus infection and dimethylnitrosamine administration on the induction of cholangiocarcinoma in Syrian hamsters

The food-borne liver trematode Opisthorchis felineus is an emerging source of biliary tract diseases on the territory of the former Soviet Union and Eastern Europe. This parasite along with trematodes Opisthorchis viverrini and Clonorchis sinensis belong to the triad of epidemiologically important liver flukes of the Opisthorchiidae family.It is known that O. viverrini and С. sinensis are the main risk factors of cholangiocarcinoma (CCA) in the endemic regions. The carcinogenic potential of О. felineus has not been well researched because of the absence of systematic pathomorphological, clinical, and epidemiological studies on O. felineus opisthorchiasis.In the present study, we show the results of detailed histopathological analysis and comprehensive evaluation of inflammation, bile duct dysplasia, periductal fibrosis, bile duct hyperplasia, bile duct proliferation, egg granuloma, cysts, cholangiofibrosis, and CCA from 10 to 30weeks following infection of Syrian hamsters with O. felineus accompanied by oral administration of dimethylnitrosamine (DMN). The results revealed that O. felineus contributes to bile duct cancer development in the hamster model. During the combined action of O. felineus and DMN, morphological features of the liver underwent dramatic changes at the cellular and organ levels. Already in the early stages of the experiment, we observed extensive periductal fibrosis, active inflammation, proliferation of the bile duct, bile duct dysplasia and egg granulomas. Later, against the background of all these changes, cholangiofibrosis and CCA were found.Our work is the first step in the study of carcinogenic potential of O. felineus. Obtained data indicate the risk of CCA of patients having chronic О. felineus opisthorchiasis, and underscore the need for the development of programs for control of this helminthiasis.

Anticancer Activity of Atractylodes lancea (Thunb.) DC in a Hamster Model and Application of PET-CT for Early Detection and Monitoring Progression of Cholangiocarcinoma.

Opisthorchis viverrini (OV)-induced cholangiocarcinoma (CCA) is an important cancer in the Great Mekong region, particularly in Thailand. Limitations of treatment options and the lack of an effective diagnostic tool for early detection of CCA are major concerns for the control of this type of cancer. The aim of the study was to investigate anti-CCA activity of the ethanolic extract of Atractylodes lancea (Thunb.) DC., and the applicability of positron emission tomography-computed tomography (PET-CT) as a tool for detection and monitoring the progression of CCA in Opisthorchis viverrini (OV)/dimethylnitrosamine (DMN)-induced CCA hamsters. Male Syrian hamsters were used for toxicity tests and anti-CCA activity evaluation. Development of CCA was induced by initial feeding of 50 metacercariae of OV, followed by drinking water containing 12.5 ppm of DMN in hamsters. The ethanolic extract of A. lancea (Thunb.) DC. was administered orally for 30 days. PET-CT was performed every 4 weeks after initiation of CCA using 18F-fluorodeoxyglucose (18F-FDG). Results from the present study suggest that the ethanolic extract of A. lancea (Thunb.) DC. rhizome exhibited promising anti-CCA activity and safety profile in the OV/DMN-induced hamster model. To successfully apply PET-CT as a tool for early detection of tumor development and progression, modification of radiolabeling approach is required to improve its specificity for CCA cells.