CCl4 Treatment Research Articles

Therapeutic implications of ajwa dates (Phoenix dactylifera) in the inhibition of liver tissue alterations through the modulation of vascular endothelial growth factor and phosphatase, and tensin homolog gene

Background: Dates are a plant species in the palm family, Arecaceae and is used as staple food in the Middle East for thousands of years. Aim: The current study was undertaken to evaluate the protective role of ajwa dates extract (ADE) on carbon tetrachloride (CCl4)-induced hepatotoxicity. Materials and Methods: The study was carried out on mice mode through different groups as control group without treatment of CCl4, ADE and CCl4-treated group, and CCl4-treated group only. Results: This finding demonstrated that histological alterations including degeneration, congestion and infiltration of lymphocytes were seen in the liver tissue in CCl4-treated groups. However, ADE-treated groups showed protection to attenuate CCl4-induced liver toxicity through maintenance of architecture of hepatocytes as evident of congestion, necrosis, and degeneration was not noticed. Moreover, a few number of infiltrations of lymphocytes were noticed in ADE-treated groups. However, expression of phosphatase and tensin homolog (PTEN) and vascular endothelial growth factor (VEGF) protein was evaluated in all groups, and it was observed that PTEN was highly expressed in control group and ADE-treated group. Whereas, PTEN expression was also observed in CCl4-treated group, but out of eight cases, one case showed less expression of PTEN protein. The difference in expression pattern of PTEN protein in CCl4-treated group and ajwa-treated group was statically insignificant (P > 0.05). VEGF was not expressed in control and ajwa-treated group. While expression of VEGF protein was observed in CCl4-treated group, and it was noticed that two out of eight cases showed expression of VEGF protein. Conclusion: The findings supported the idea that ADE might reduce liver tissue alterations or maintenance of architecture of hepatocytes. Ajwa dates-treated group showed decreased in the VEGF expression and such angiogenesis process involve in migration and differentiation of endothelial cells as well as prevent the loss of PTEN protein expression. Abbreviations Used: CCl4: Carbon Tetrachloride, VEGF: Vascular Endothelial Growth factor, IHC: Immunohistochemistry TUNEL assay: Terminal deoxynucleotidyl transferase dUTP nick end labeling, H and E: Hematoxylin-eosin staining, ADE: Ajwa dates extract.

Stevia Prevents Acute and Chronic Liver Injury Induced by Carbon Tetrachloride by Blocking Oxidative Stress through Nrf2 Upregulation.

The effect of stevia on liver cirrhosis has not been previously investigated. In the present study, the antioxidant and anti-inflammatory properties of stevia leaves were studied in male Wistar rats with carbon tetrachloride- (CCl4-) induced acute and chronic liver damage. Acute and chronic liver damage induced oxidative stress, necrosis, and cholestasis, which were significantly ameliorated by stevia. Chronic CCl4 treatment resulted in liver cirrhosis, as evidenced by nodules of hepatocytes surrounded by thick bands of collagen and distortion of the hepatic architecture, and stevia significantly prevented these alterations. Subsequently, the underlying mechanism of action of the plant was analyzed. Our study for the first time shows that stevia upregulated Nrf2, thereby counteracting oxidative stress, and prevented necrosis and cholestasis through modulation of the main proinflammatory cytokines via NF-κB inhibition. These multitarget mechanisms led to the prevention of experimental cirrhosis. Given the reasonable safety profile of stevia, our results indicated that it may be useful for the clinical treatment of acute and chronic liver diseases.

Gankyrin Promotes Tumor-Suppressor Protein Degradation to Drive Hepatocyte Proliferation.

Background & AimsUncontrolled liver proliferation is a key characteristic of liver cancer; however, the mechanisms by which this occurs are not well understood. Elucidation of these mechanisms is necessary for the development of better therapy. The oncogene Gankyrin (Gank) is overexpressed in both hepatocellular carcinoma and hepatoblastoma. The aim of this work was to determine the role of Gank in liver proliferation and elucidate the mechanism by which Gank promotes liver proliferation.MethodsWe generated Gank liver-specific knock-out (GLKO) mice and examined liver biology and proliferation after surgical resection and liver injury.ResultsGlobal profiling of gene expression in GLKO mice showed significant changes in pathways involved in liver cancer and proliferation. Investigations of liver proliferation after partial hepatectomy and CCl4 treatment showed that GLKO mice have dramatically inhibited proliferation of hepatocytes at early stages after surgery and injury. In control LoxP mice, liver proliferation was characterized by Gank-mediated reduction of tumor-suppressor proteins (TSPs). The failure of GLKO hepatocytes to proliferate is associated with a lack of down-regulation of these proteins. Surprisingly, we found that hepatic progenitor cells of GLKO mice start proliferation at later stages and restore the original size of the liver at 14 days after partial hepatectomy. To examine the proliferative activities of Gank in cancer cells, we used a small molecule, cjoc42, to inhibit interactions of Gank with the 26S proteasome. These studies showed that Gank triggers degradation of TSPs and that cjoc42-mediated inhibition of Gank increases levels of TSPs and inhibits proliferation of cancer cells.ConclusionsThese studies show that Gank promotes hepatocyte proliferation by elimination of TSPs. This work provides background for the development of Gank-mediated therapy for the treatment of liver cancer. RNA sequencing data can be accessed in the NCBI Gene Expression Omnibus: GSE104395.

Hepatoprotective effect of fermented black radish (Raphanus sativus L. var niger) in CCl4 induced liver injury in rats

Oxidative stress is one of common cause of fatty changes in the liver. Antioxidant capacity was confirmed in various vegetables including black radish (Raphanus sativus L. var niger). Fermentation of vegetables using Lactobacillus plantarum has been known to generate bioactive components. This study was conducted to determine if fermented black radish (FBR) ameliorates oxidative liver injury induced by CCl4 in rats. To accomplish this, FBR (250 and 500 mg/ kg) was orally administered to rats for 7 consecutive days, single CCl4 (1.5 mL/kg) treatment or no treatment orally. Serum chemistry at 24 hours after CCl4 injury showed that FBR (500 mg/kg) significantly reduced the level of both alanine aminotransferase and aspartate aminotransferase in CCl4 exposed rats. Moreover, FBR treatment significantly increased radical-scavenging effects in livers with the reduction of lipid peroxidation in CCl4 exposed rats. Histopathologic findings including Kupffer cell activation in the liver of each group matched those of serum chemistry. Collectively, black radish, through fermentation, exerts hepatoprotective capacity in CCl4 induced liver injury in rats through anti-oxidation.

Antifibrotic effects of D-limonene (5(1-methyl-4-[1-methylethenyl]) cyclohexane) in CCl4 induced liver toxicity in Wistar rats.

This study was designed to assess the potential antifibrotic effect of D-Limonene-a component of volatile oils extracted from citrus plants. D-limonene is reported to have numerous therapeutic properties. CCl4 -intduced model of liver fibrosis in Wistar rats is most widely used model to study chemopreventive studies. CCl4 -intoxication significantly increased serum aminotransferases and total cholesterol these effects were prevented by cotreatment with D-Limonene. Also, CCl4 -intoxication caused depletion of glutathione and other antioxidant enzymes while D-Limonene preserved them within normal values. Hydroxyproline and malondialdehyde content was increased markedly by CCl4 treatment while D-Limonene prevented these alterations. Levels of TNF-α, TGF-β, and α-SMA were also assessed; CCl4 increased the expression of α-SMA, NF-κB and other downstream inflammatory cascade while D-Limonene co-treatment inhibited them. Collectively these findings indicate that D-Limonene possesses potent antifibrotic effect which may be attributed to its antioxidant and anti-inflammatory properties.

Hepatoprotective effect of food preservatives (butylated hydroxyanisole, butylated hydroxytoluene) on carbon tetrachloride-induced hepatotoxicity in rat.

Carbon tetrachloride (CCl4), a hepatotoxic agent is widely used to study the toxic mechanisms in experimental animals. This study was carried out to establish the hepatoprotective measures of food preservative antioxidants butylated hydroxyanisole and butylated hydroxytolune (BHA, BHT) when mixed with food towards carbon tetrachloride (CCl4) intoxication (230mg/kg b wt/rat/day) in rat. Biochemical markers like serum glutamate pyruvate tranaminase (AST), serum glutamate oxaloacetate transaminase (ALT), alkaline phosphatase (ALP) and bilirubin content, antioxidant enzymes such as SOD, CAT, GPx, and malondialdehyde (MDA) as the end product of lipid peroxidanion were measured. The results had shown the elevated level of AST (121.16%), ALT (124.68%), ALP (122.41%) an, bilirubin content (57.14%) after CCl4 treatment. Marked decrease of activity of antioxidant enzymes such as SOD (85.36%), CAT (67.47%), GPx (50.7%) had indicated that the ROS mediated toxicity and pretreatment of BHA and BHT restored the activity of these enzymes. High level of MDA content with reduced GSH value was also observed due to oxidative stress. The hepatic antioxidant status was restored with the food preservative (BHA, BHT) antioxidant treatment which had indicated the significant protective effect against CCl4 induced hepatotoxicity and finally confirmed by histopathological studies.

Lycium barbarum polysaccharides improve CCl4-induced liver fibrosis, inflammatory response and TLRs/NF-kB signaling pathway expression in wistar rats

Lycium barbarum polysaccharides (LBPs) have multiple biological and pharmacological functions, including antioxidant, anti-inflammatory and anticancer activities. This research was conducted to evaluate whether LBPs could alleviate carbon tetrachloride (CCl4)-induced liver fibrosis and the underlying signaling pathway mechanism. Fifty male wistar rats were randomly allocated to five groups (n=10): control, CCl4 and CCl4 with 400, 800 or 1600mg/kg LBPs, respectively. Each wistar rat from each group was used for blood and tissue collections at the end of experiment. The results showed that CCl4 induced liver fibrosis as demonstrated by increasing histopathological damage, α-smooth muscle actin expression, aspartate transaminase activities, alkaline phosphatase activities and alanine aminotransferase activities. LBPs supplementation alleviated CCl4-induced liver fibrosis as demonstrated by reversing the above parameters. In addition, CCl4 treatment induced the oxidative injury, increased the mRNA levels of tumor necrosis factor-α, monocyte chemoattractant protein-1 and interleukin-1β, and up-regulated the protein expressions of toll-like receptor 4 (TLR4), TLR2, myeloid differentiation factor 88, nuclear factor-kappa B (NF-kB) and p-p65. LBPs supplementation alleviated CCl4-induced oxidative injury, inflammatory response and TLRs/NF-kB signaling pathway expression by reversing the above some parameters. These results suggest that the alleviating effects of LBPs on CCl4-induced liver fibrosis in wistar rats may be through inhibiting the TLRs/NF-kB signaling pathway expression.

MiR-185 Inhibits Fibrogenic Activation of Hepatic Stellate Cells and Prevents Liver Fibrosis.

Recent studies have shown the effect of microRNAs on HSC activation and transformation, which is essential for the pathogenesis of liver fibrosis. In our study, we explored the role of miR-185 in liver fibrosis. Plasma miR-185 was detected in hepatitis B virus-related liver fibrosis patients (S2/3, n = 10) by Illumina HiSeq sequencing, and healthy volunteers were selected (n = 8) as the control group. We found that the plasma miR-185 level in fibrosis patients was significantly downregulated. CCl4-induced fibrosis tissues in mouse livers and TGF-β1-activated HSCs also presented downregulated miR-185 concomitant with an increased expression of RHEB and RICTOR. To explore the correlations, LX-2 cells were transiently transfected with miR-185 mimics. The expression levels of α-SMA, collagen I, and collagen III were decreased as well as RHEB and RICTOR. Inhibition of endogenous miR-185 increased fibrogenic activity. Furthermore, dual-luciferase reporter assays indicated that miR-185 inhibited the expression of RHEB and RICTOR by directly targeting their 3′ UTRs. Moreover, silencing RHEB and RICTOR suppressed α-SMA and collagen expression levels. In conclusion, miR-185 prevents liver fibrogenesis by inhibiting HSC activation via inhibition of RHEB and RICTOR. These results provide new insights into the mechanisms behind the anti-fibrotic effect of miR-185.

CM-101: Type I Collagen-targeted MR Imaging Probe for Detection of Liver Fibrosis.

Purpose To evaluate the biodistribution, metabolism, and pharmacokinetics of a new type I collagen-targeted magnetic resonance (MR) probe, CM-101, and to assess its ability to help quantify liver fibrosis in animal models. Materials and Methods Biodistribution, pharmacokinetics, and stability of CM-101 in rats were measured with mass spectrometry. Bile duct-ligated (BDL) and sham-treated rats were imaged 19 days after the procedure by using a 1.5-T clinical MR imaging unit. Mice were treated with carbon tetrachloride (CCl4) or with vehicle two times a week for 10 weeks and were imaged with a 7.0-T preclinical MR imaging unit at baseline and 1 week after the last CCl4 treatment. Animals were imaged before and after injection of 10 µmol/kg CM-101. Change in contrast-to-noise ratio (ΔCNR) between liver and muscle tissue after CM-101 injection was used to quantify liver fibrosis. Liver tissue was analyzed for Sirius Red staining and hydroxyproline content. The institutional subcommittee for research animal care approved all in vivo procedures. Results CM-101 demonstrated rapid blood clearance (half-life = 6.8 minutes ± 2.4) and predominately renal elimination in rats. Biodistribution showed low tissue gadolinium levels at 24 hours (<3.9% injected dose [ID]/g ± 0.6) and 10-fold lower levels at 14 days (<0.33% ID/g ± 12) after CM-101 injection with negligible accumulation in bone (0.07% ID/g ± 0.02 and 0.010% ID/g ± 0.004 at 1 and 14 days, respectively). ΔCNR was significantly (P < .001) higher in BDL rats (13.6 ± 3.2) than in sham-treated rats (5.7 ± 4.2) and in the CCl4-treated mice (18.3 ± 6.5) compared with baseline values (5.2 ± 1.0). Conclusion CM-101 demonstrated fast blood clearance and whole-body elimination, negligible accumulation of gadolinium in bone or tissue, and robust detection of fibrosis in rat BDL and mouse CCl4 models of liver fibrosis. © RSNA, 2017 Online supplemental material is available for this article.

Pericentral hepatocytes produce insulin-like growth factor-2 to promote liver regeneration during selected injuries in mice.

IGF-2 is produced by pericentral hepatocytes to promote hepatocyte proliferation and repair tissue damage in the setting of chronic liver injury, which is distinct from the signaling that occurs post-PHx. (Hepatology 2017;66:2002-2015).

Xiaochaihutang attenuates liver fibrosis by activation of Nrf2 pathway in rats

Xiaochaihutang (XCHT) is a decoction of seven botanical extracts used for liver diseases traditionally in East Asia. However, few studies have investigated its anti-hepatic fibrosis effects and the mechanisms of action. Oxidative stress is one of the key factors responsible for occurrence and development of hepatic fibrosis and nuclear factor erythroid 2-related factor 2 (Nrf2) serves as a major regulator of the cellular defense system against oxidative stress. The aim of the present study was to investigate the effect of XCHT on liver fibrosis and focuse on the Nrf2 pathway in the protection of XCHT against CCl4-induced oxidative stress. Liver fibrosis was induced by repeated injection of CCl4 over a period of 9 weeks. Starting from the 6th week, rats in treatment groups were given the appropriate doses of XCHT granules and Silybin. We discovered that CCl4 caused significant fibrosis damage in rat liver, and XCHT (5g/kg and 10g/kg, po for 4 weeks) significantly improved the liver functions and fibrosis degree. XCHT treatment significantly decreased the number of α-SMA positive stained cells, indicating suppression of activated hepatic stellate cells (HSCs). Compared with the CCl4 treatment, administration of XCHT significantly increased the hepatic levels of Nqo1, HO-1, GCLC and GCLM, the major components of the Nrf2 pathway. These studies demonstrated that XCHT is an effective therapeutic agent for treatment of hepatic fibrosis and the mechanism of action might be due to up-regulation of the Nrf2 pathway against oxidative stress, making further inhibition of activated HSCs.

Quercetin reverses experimental cirrhosis by immunomodulation of the proinflammatory and profibrotic processes.

The ability of quercetin to reverse an established cirrhosis has not yet been investigated. Therefore, the aim of this study was to examine the efficacy of this flavonoid in reversing experimental cirrhosis. Cirrhosis was induced by intraperitoneal administration of TAA (200mg/kg of body weight) three times per week for 8weeks or by intraperitoneal petrolatum-CCl4 (400mg/kg of body weight) administration three times per week for 8weeks. To determine the capacity of quercetin to prevent liver fibrosis, the flavonoid (50mg/kg of body weight, p.o.) was administered daily to rats during the CCl4 or TAA treatment. To evaluate the ability of quercetin to reverse the previously induced cirrhosis, we first treated rats with CCl4 for 8weeks, as previously described and then the flavonoid was administered for four more weeks. We found that the liver anti-inflammatory and antinecrotic effects of quercetin are associated with its antioxidant properties, to the ability of the flavonoid to block NF-κB activation and in consequence to reduce cytokine IL-1. The ability of quercetin to reverse fibrosis may be associated with the capacity of the flavonoid to decrease TGF-β levels, hepatic stellate cell activation, and to promote degradation of the ECM by increasing metalloproteinases. The main conclusion is that quercetin, in addition to its liver protective activity against TAA chronic intoxication, is also capable of reversing a well-stablished cirrhosis by blocking the prooxidant processes and by downregulating the inflammatory and profibrotic responses.

Does Eryngium maritimum seeds extract protect against CCl4 and cisplatin induced toxicity in rats: Preliminary phytochemical screening and assessment of its in vitro and in vivo antioxidant activity and antifibrotic effect

The present work was undertaken to investigate the protective effects of Eryngium maritimum seeds extract (MEE) against CCl4 and cisplatin induced toxicity in rats. The levels of total phenolics and flavonoids were ca. 20mg GAE/g and ca. 19mg QE/g, respectively. Using HPLC twenty-one molecules were identified. MEE showed important in vitro antioxidant activities. Results demonstrated that MEE-treatment restored the increased activities of AST, ALT, LDH and ALP and the high levels of serum creatinine, urea and uric acid levels induced by CCl4 and cisplatin treatment. Moreover, MEE presented important in vivo antioxidant activities; increased the activities of CAT, SOD and GPx and decreased the TBARS and protein carbonyl contents. The histopathological studies suggested that MEE clearly alleviated the degree of tissues fibrosis. The obtained results highlighted the potential use of E. maritimum as a source of bioactive compounds with hepatoprotective and nephroprotective advantages.

Phytochemical investigation and nephroprotective potential of Sida cordata in rat

BackgroundPlants are an efficient source of natural antioxidant against free radicals causing kidney damages. Sida cordata ethyl acetate fraction has been reported for strong in vitro antioxidant potency, previously. In the present study, our objective was to evaluate its in vivo antioxidant potency against CCl4 induced nephrotoxicity and investigates the bioactive phytochemicals by HPLC-DAD analysis.MethodsPhytochemical analysis was performed by HPLC-DAD methodology. For in vivo study, 42 male Sprague-Dawley rats were treated with alternatively managed doses for 60 days. Group I animals were remained untreated. Group II animals were treated with vehicle (1 mL of olive oil) by intragastric route on alternate days. Group III was treated with 30% CCl4 (1 mL/kg b.w.) i.p. Group IV was treated with 30% CCl4 (1 mL/kg b.w.) i.p and silymarin intragastric. Group V and VI rats were treated with 30% CCl4 and SCEE (150 and 300 mg/kg b.w., respectively) intragastric. Group VII animals were treated with SCEE (300 mg/kg b.w.) intragastrically. Blood parameters, Serum proteins and urine profile were investigated. Activities of tissue enzyme i.e. catalase, peroxidase, superoxide dismutase, glutathione-S-transferase, glutathione reductase, GSH and γ-GT were evaluated. Histopathological observations, total protein contents, lipid peroxidation, DNA damage and relative weight were also analyzed.ResultsGallic acid, catechin and caffeic acid were identified in SCEE fraction by HPLC-DAD. Decrease in the count of red blood cells, neutrophils, eosinophils and concentration of hemoglobin whereas increase in lymphocyte count and estimation of sedimentation rate (ESR) with 1 mL CCl4 (30% in Olive oil) administration (30 doses in 60 days) was restored dose dependently with co-treatment of SCEE (150 and 300 mg/kg b.w.). Treatment of rats with CCl4 markedly (P < 0.01) increased the count of urinary red blood cells and leucocytes, concentration of urea, creatinine and urobilinogen and specific gravity whereas creatinine clearance was reduced. Serum level of total protein, albumin, globulin, nitrite, creatinine and blood urea nitrogen (BUN) was significantly increased (P < 0.01) by CCl4 treatment. The activity of antioxidant enzymes; catalase, superoxide dismutase, glutathione peroxidase, glutathione-S-transferase and glutathione reductase and content of reduced glutathione was decreased (P < 0.01) significantly. However, increased concentration (P < 0.01) of thiobarbituric acid reactive substances and histopathological injuries were noticed in the renal tissues of rats after the treatment with CCl4. Co-administration of SCEE, dose dependently, protected the alterations in the studied parameters of rats at 150 and 300 mg/kg b.w. The present study revealed that SCEE could be used as a possible remedy for renal toxicity abnormalities.ConclusionThese results are an evidence of the renal protective role of S.cordat ethyl acetate fraction against CCl4 induced nephrotoxicity in rats which may be due to its antioxidant compounds.

Curcumin and Curcuma longa L. extract ameliorate lipid accumulation through the regulation of the endoplasmic reticulum redox and ER stress

For this study, we examined the effects of curcumin against acute and chronic stress, paying specific attention to ROS. We also aimed to clarify the differences between acute and chronic stress conditions. We investigated the effects of curcumin against acute stress (once/1 day CCl4 treatment) and chronic-stress (every other day/4week CCl4 treatment). Compared with acute stress, in which the antioxidant system functioned properly and aspartate transaminase (AST) and ROS production increased, chronic stress increased AST, alanine aminotransferase (ALT), hepatic enzymes, and ROS more significantly, and the antioxidant system became impaired. We also found that ER-originated ROS accumulated in the chronic model, another difference between the two conditions. ER stress was induced consistently, and oxidative intra-ER protein folding status, representatively PDI, was impaired, especially in chronic stress. The PDI-associated client protein hepatic apoB accumulated with the PDI-binding status in chronic stress, and curcumin recovered the altered ER folding status, regulating ER stress and the resultant hepatic dyslipidemia. Throughout this study, curcumin and curcumin-rich Curcuma longa L. extract promoted recovery from CCl4-induced hepatic toxicity in both stress conditions. For both stress-associated hepatic dyslipidemia, curcumin and Curcuma longa L. extract might be recommendable to recover liver activity.

Elevated levels of liver methylglyoxal and d-lactate in early-stage hepatitis in rats.

Methylglyoxal (MGO) is highly cytotoxic and its levels are elevated in diabetes, nephropathy and atherosclerosis. However, it has never been studied in liver disease. For this reason, we aimed to assess the levels of MGO and its metabolite d-lactate in an early hepatitis model. Wistar rats were administered CCl4 (0.75 mL/kg, i.p.) to induce hepatitis. In either CCl4 -treated or untreated rats, alanine transaminase and aspartate transaminase levels did not change over the course of the study, indicating that significant liver damage did not occur following CCl4 treatment. However, the levels of MGO and d-lactate were higher in the livers of CCl4 -treated animals than in untreated animals (MGO: 128.2 ± 18.8 and 248.1 ± 64.9 μg/g protein, p < 0.01; d-lactate: 0.860 ± 0.040 and 1.293 ± 0.078 μmol/g protein, respectively p < 0.01). Furthermore, in untreated and treated animals, serum d-lactate levels were 57.65 ± 2.59 and 92.16 ± 16.69 μm and urine d-lactate levels were 1.060 ± 0.007 and 1.555 ± 0.366 μmol/mg UCr, respectively (p < 0.01). These data show that in this model of early-stage liver damage, the levels of MGO and its metabolite d-lactate are elevated and that d-lactate could be useful as a reference marker for the early stage of hepatitis.

Effects of exogenous thymosin \u03b24 on carbon tetrachloride-induced liver injury and fibrosis

The present study investigated the effects of exogenous thymosin β4 (TB4) on carbon tetrachloride (CCl4)-induced acute liver injury and fibrosis in rodent animals. Results showed that both in mice and rats CCl4 rendered significant increases in serum alanine aminotransferase and aspartate aminotransferase, hepatic malondialdehyde formation, decreases in antioxidants including superoxide dismutase and glutathione, and up-regulated expressions of transforming growth factor-β1, α-smooth muscle actin, tumor necrosis factor-α and interleukin-1β in the liver tissues. Hydroxyproline contents in the rat livers were increased by CCl4. Histopathological examinations indicated that CCl4 induced extensive necrosis in mice livers and pseudo-lobule formations, collagen deposition in rats livers. However, all these changes in mice and rats were significantly attenuated by exogenous TB4 treatment. Furthermore, up-regulations of nuclear factor-κB p65 protein expression by CCl4 treatment in mice and rats livers were also remarkably reduced by exogenous TB4 administration. Taken together, findings in this study suggested that exogenous TB4 might prevent CCl4-induced acute liver injury and subsequent fibrosis through alleviating oxidative stress and inflammation.

Branched chain amino acids supplementation modulates TGF-β1/Smad signaling pathway and interleukins in CCl4 -induced liver fibrosis.

The alterations and low levels of circulating branched chain amino acids (BCAAs), leucine, isoleucine, and valine, are associated with liver diseases. The study was designed to evaluate hepatoprotective effect of BCAAs on CCl4 -induced liver fibrosis and to investigate the molecular mechanisms underlying these effects in rats. In all, 30 male rats were divided into three groups. Control group (n = 10) and CCl4 group (n = 10), where rats were injected with CCl4 (1 mL/kg of 0.5 : 1 v/v injected i.p. twice weekly for 12 weeks). In CCl4 + BCAAs group (n = 10), rats were injected with similar doses of CCl4 and supplemented with a mixture of 600 mg/kg BCAAs (2 : 1 : 1.2 leucine : isoleucine : valine) by oral gavage, three times/week for 12 weeks. Liver fibrosis was assessed by measuring total bilirubin, total protein, alanine aminotransferase, and aspartate aminotransferase, hydroxyproline content, and serum IL-6 and IL-10. Histopathologic studies and α-smooth muscle actin (α-SMA) were detected immunohistochemically in liver. Serum insulin level, blood glucose, liver malodialdehyde concentration (MDA), glutathione peroxidase, and superoxide dismutase (SOD) activities were quantified. TGF-β1, Smad3, and Smad7 gene expressions were estimated by qRT-PCR. BCAAs suppressed liver fibrosis induced by CCl4 treatment. BCAAs modulated liver indices and downregulated TGF-β1, Smad3, and Smad7 expressions in hepatocytes. BCAAs enhanced liver antioxidant enzyme activities (P < 0.001), reduced serum levels of TGF-β1, IL-6, and IL-10 compared to CCL4 group and ameliorated histopathologic changes in rat liver. BCAAs may have a protective role against liver fibrosis via antioxidant and anti-inflammatory mechanisms.

Bone marrow endothelial progenitor cells activate hepatic stellate cells and aggravate carbon tetrachloride induced liver fibrosis in mice via paracrine factors.

Bone marrow derived endothelial progenitor cells (BM-EPCs) are increased in chronic liver disease (CLD). Their role in hepatic fibrosis and regeneration remains an area of intense studies. We investigated the migration and secretory functions of BM-EPCs in fibrotic mice liver. Bone marrow cells from C57BL6-GFP mice were transplanted into the femur of irradiated C57BL6 mice, followed by CCl4 doses for 8 weeks, to develop hepatic fibrosis (n=36). Transplanted C57BL6 mice without CCl4 treatment were used as controls. EPCs were analyzed in BM, blood and liver by flow cytometry and immunofluorescence. VEGF and TGF-β were analysed in the hepatic stellate cells (HSCs) and BM-EPCs co-cultures using ELISAs. There was a significant migration of EPCs from BM to blood and to the liver (P≤0.01). Percentage of GFP+ CD31+ EPCs and collagen proportionate area was substantially increased in the liver at 4th week of CCl4 dosage compared to the controls (19.8% vs 1.9%, P≤0.05). Levels of VEGF (533.6pg/ml) and TGF-β (327.44pg/ml) also increased significantly, when HSCs were treated with the EPC conditioned medium, as compared to controls (25.66pg/ml and 5.87pg/ml, respectively; P≤0.001). Present findings suggest that BM-EPCs migrate to the liver during CCl4-induced liver injury and contribute to fibrosis.

Cost-analysis and effectiveness of one-stage laparoscopic versus two-stage endolaparoscopic management of cholecystocholedocholithiasis: a retrospective cohort study

BackgroundOne–stage laparoscopic common bile duct (CBD) stone clearance and laparoscopic cholecystectomy (LCBDE+LC) for cholecystocholedocholithiasis ( CCL) can be performed with similar short and long-term outcomes than two-stage endoscopic retrograde cholangiography followed by subsequent LC (ERCP+LC). This study examined retrospectively the outcome and hospital costs of one-stage versus two-stage treatment of CBD stones.MethodsFrom January 1999 and December 2014, 217 consecutive, elective patients underwent one-stage (LCBDE + LC ) or two-stage (ERCP + subsequent LC ) treatment for CBD stones. The data from the one-stage management was collected prospectively, and from the two-stage management retrospectively. The main measure of outcome was hospital costs, with the success of one-stage versus two-stage management, postoperative morbidity and postoperative stay as secondary outcome measures.ResultsOne-stage laparoscopic transcystic management was the least costly option compared to laparoscopic one-stage transductal approach (TC 5455€ versus TD 9364, p < 0.001) or two-stage management (6913 €, p = 0.02). Overall success rate of primary intervention (including conversions to open surgery) for CBD stone clearance was 96.9%, 97.0% and 98.3% after transcystic one-stage, transductal one-stage and two-stage approach, p = 0.79. Postoperative morbidity was 15.5% versus 7.5%, p = 0.64, and postoperative hospital stay median 2 days (IQR 2–5) versus 4.5 days ( IQR 3–7), p < 0.001 in the one-stage and two-stage management groups.ConclusionsOur study shows that laparoscopic one-stage transcystic management of CCL results in high rate of CBD clearance, fewer procedures per patient, shorter hospital and lower costs than the two-stage management. Therefore the one-stage transcystic management seems to be an attractive strategy for the treatment of CCL depending on local resources and surgical expertise .