Cirrhosis In Rats Research Articles

The Effects of the “XGTQ” Medicine in Treating Cirrhosis in Wistar Rats

Objectives: To evaluate the effects of “XGTQ” in the treatment of cirrhosis induced by Carbon tetrachloride (CCL4) in combination with alcohol and a high-fat diet on Wistar rats. Materials and methods: Cirrhosis on white rats was induced by subcutaneously injecting CC14 at an initial dose of 5,0ml/kg, followed by 1,2ml/kg once a week in 10 weeks. Then, fed with synthetic food, added 20% fat, and 0.05% cholesterol and iron oxalate. Rats were administered every day with plain water and 1 day with water mixed with 30% ethanol. The rats were randomly divided into 5 groups and given distilled water (group 1 and 2 or control group), silymarin (group 3 or reference group) or the “XGTQ” drug extract (group 4, 5) for 4 weeks. Collected blood for biochemical test and liver were dissected to evaluate weight, morphology, and quantified 4-hydroxyproline to evaluate fibrosis and collagen accumulation. Results: n cirrhotic Wistar rats, “XGTQ” drug at 19.6 g/kg/24h and 58.8 g/kg/24h showed the ability toreduce the activity of enzymes AST, ALT in the blood (p<0.01), increasing plasma albumin and decreasing prothrombin time (p<0.05); improving physical condition, macroscopic and microscopic images of H&E-stained liver; decreasing the concentration of hydroxyproline in the liver and reducing the level of cirrhosis on the Masson-stained templates. The effects of “XGTQ” increased with the dose and was equivalent to silymarin at the dose of 70 mg/kg/24h. Conclusion: The extract of “XGTQ” drug is effective in treating cirrhosis in Wistar rats.

1-Methyl tryptophan, an indoleamine 2,3-dioxygenase inhibitor, attenuates cardiac and hepatic dysfunction in rats with biliary cirrhosis

Kynurenine Pathway (KP) is the dominant metabolic route of tryptophan which is catalyzed by indoleamine-2,3-dioxygenase (IDO). This pathway is upregulated in liver disease where the level of KP metabolites correlates with the severity of disease. Cirrhosis is associated with cardiac dysfunction, which manifests itself during severe physiological challenges such as liver transplantation. Cardiac dysfunction in cirrhosis is linked to systemic inflammation and impaired cardiac beta-adrenergic signaling pathways. The KP pathway is involved in modulation of cardiac signaling and is upregulated by systemic inflammation. Therefore, this study aimed to evaluate the effect of IDO inhibition on development of cardiac dysfunction in an experimental model of cirrhosis. Cirrhosis was induced by bile duct ligation (BDL). Experimental groups were given either 1-methyl tryptophan (1-MT, 1, 3, 9 mg/kg), or saline. 28 days after BDL, cardiac chronotropic response to epinephrine was assessed ex vivo. HPLC was employed to measure hepatic and cardiac levels of tryptophan, kynurenine and kynurenic acid. Cirrhosis in rats was associated with impaired cardiac chronotropic responsiveness to adrenergic stimulation. 1-MT dose-dependently improved cirrhosis-induced chronotropic dysfunction as well as elevated serum levels of CRP and IL-6 in BDL rats. Hepatic and cardiac kynurenine/tryptophan ratio were elevated in cirrhotic rats and were reduced following 1-MT administration. Chronic administration of 1-MT could also reduce hepatic inflammation, fibrosis and ductular proliferation. 1-MT attenuates cardiac dysfunction in rats with biliary cirrhosis. This protective effect is not limited to the cardiac function as liver histopathologic changes were also improved following chronic 1-MT administration.

The correlation between magnetic resonance diffusion parameters and Ki-67 and PCNA in hepatic fibrosis and cirrhosis rats.

The aim of this study was to assess the value of different 1.5 T MRI apparent diffusion coefficient (ADC) and exponential apparent diffusion coefficient (EADC) values in diagnosing the stages of liver cirrhosis. Sprague-Dawley (SD) rats were randomly divided into the experimental group and the control group. The experimental group was injected with thioacetamide intraperitoneally 3 times per week. After routine MR scanning, diffusion-weighted imaging (DWI) was processed by spin echo-echo planar imaging (SE-EPI) to generate the ADC value and EADC image. The liver ADC and EADC values of rats were measured in the control and experimental groups, followed by Masson staining and hematoxylin and eosin staining. Furthermore, immunohistochemistry was performed to detect Ki-67 and PCNA expression in liver tissues. In the control group, the differences in ADC and EADC values between the liver fibrosis and cirrhosis group were different. The ADC values of the liver fibrosis stage I-II, III-IV, and cirrhosis rats in the experimental group were lower than the control group, while the EADC values were higher than the control group. The ADC values of the liver fibrosis stage III-IV group and cirrhosis nodules group were lower than the control group. There were significant differences in EADC values between the cirrhotic nodule groups and the control group. DWI-ADC values showed a negative correlation between SD rat liver fibrosis and cirrhosis pathology classification.

Mitoquinol Mesylate Ameliorates Hematological Aberration in Cirrhotic-Hepatocellular Carcinogenic Rats

Introduction: Hematological abnormalities such as anemia, neutropenia, and thrombocytopenia experienced by cancer patients undergoing chemotherapy necessitate the need to develop agents that stimulate erythropoiesis and boost immune response in cancer. Aim: The present study investigated the effects of mitoquinol mesylate (MitoQ) on the hematological profile of diethyl nitrosamine (DEN)-induced cirrhotic-hepatocellular carcinogenic (HCC) rats. Materials and Methods: One hundred Wistar strain albino rats were randomly divided into five groups (n = 20) for the experimental period of 20 weeks. Groups A, B, and C received distilled water, 10 mg/kg each of DEN and MitoQ respectively. Animals in Group D were pretreated with 10 mg/kg MitoQ for a week followed by coadministration of 10 mg/kg each of MitoQ and DEN for 20 weeks, while Group E received 10 mg/kg DEN for 8 weeks and then coadministration of 10 mg/kg each of DEN and MitoQ till the end of 20th week. Animals were sacrificed at the end of 12th, 16th, and 20th week. Blood samples were collected into labeled heparinized bottles for hematological analysis. Results: Administration of DEN significantly (P < 0.05) decreased the levels of red blood cell (RBC) count and erythrocytes indices. These erythrogram parameters were continuously deranging from 6.86 ± 0.26 to 2.69 ± 0.01, from 37.40 ± 1.99 to 30.35 ± 0.57, from 12.16 ± 0.97 to 8.90 ± 0.37 in the levels of RBC, packed cell volume, and hemoglobin, respectively, with the progression of HCC from cirrhosis to advanced HCC in Wistar rats. Further, the levels of white blood cells (WBCs, 3.68 ± 0.08), granulocytes (1.30 ± 0.06), lymphocytes (0.63 ± 0.11), and platelets (440.00 ± 22.67) were significantly reduced in the DEN group compared to healthy control (4.46 ± 0.12, 2.14 ± 0.27, 1.70 ± 0.13, and 685.80 ± 15.48 in the levels of WBC, granulocytes, lymphocytes, and platelets, respectively) at the advanced HCC stage. Interestingly, MitoQ interventions significantly (P < 0.05) reversed alterations in hematological indices induced by DEN toxicity. Conclusion: Our findings suggest that MitoQ is safe and capable of normalizing hematological abnormalities associated with cirrhosis and HCC in Wistar rats.

РОЛЬ УРОВНЯ МРНК ГЕНОВ СИГНАЛЬНОГО ПУТИ NOTCH ПРИ ИНДУЦИРОВАННОМ ФИБРОГЕНЕЗЕ ПЕЧЕНИ КРЫСЫ

Objectives. To study the role of mRNA level of the Notch signaling pathway genes in induced rat liver fibrogenesis. Material and methods. Fibrosis followed by the transition to liver cirrhosis in rats of Wistar line was induced with thioacetamide at a dose of 200 mg/kg of animal body weight twice a week for 17 weeks. The rats were randomized into 9 groups of 12 animals each. The mRNA level of the Notch signaling pathway genes was assessed by real-time PCR. The notch1, notch2, yap1 and hes1 genes were used as molecular targets. Microscopic analysis of histological preparations was performed using the OLYMPUS BX51 microscope. The degree of fibrosis was assessed according to the scale of Ishak K.G. Results. The study of the classical transcription factor of the Notch signaling pathway, hes1, revealed its very low and stable activity in all studied samples. The analysis of relative dynamics of the mRNA level of the notch1, notch2, and yap1 genes made it possible to determine marked changes in their levels at the point of transition from the normal state of liver tissues to the development of fibrosis. Conclusions. Within the framework of this study, the hes1 gene is not a target of the Notch pathway and can be used as a reference gene. The noted decrease in the mRNA level of the yap1 gene, probably, inhibits the compensatory-restorative processes in the liver, activates the stellate cells, and promotes the transformation of fibrosis into cirrhosis. In addition, it has been found that the revealed fluctuations in the mRNA levels of the notch1 and yap1 genes in relation to the starting point (there are no changes in the liver tissue) quite accurately describe the period of the onset of the transition of advanced fibrosis to cirrhosis. In this regard, they can be considered as potential markers of the transition of fibrosis to cirrhosis.

Effects of probiotics on intestinal flora, inflammation and degree of liver cirrhosis in rats with liver cirrhosis by regulating Wnt/β-catenin signaling pathway.

We aimed to explore the effects of probiotics on intestinal flora, inflammation and degree of liver cirrhosis in rats with liver cirrhosis, and to verify the Wnt/β-catenin signaling pathway that regulates this process. A total of 30 SD rats were randomly divided into 3 groups, namely, control group (n=10), model group (n=10) and probiotic group (n=10). Rats in the model group were used to construct liver cirrhosis models using carbon tetrachloride (CCL4) method, and those in the probiotic group were administered with probiotic preparations by gavage for 8 weeks. Then the feces of rats in each group were taken to detect the composition of intestinal flora, and changes in the content of inflammatory cytokines, such as tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), monocyte chemotactic protein 1 (MCP-1) and interferon-gamma (IFN-γ), in peripheral blood collected were examined by enzyme-linked immunosorbent assay (ELISA). Next, changes in the degree of liver cirrhosis were analyzed by hematoxylin and eosin (H&E) staining, and the expression levels of the Wnt/β-catenin signaling pathway-related molecules, including β-catenin, glycogen synthase kinase (GSK)-3β and Frizzled-2, in liver tissues in each group were detected via polymerase chain reaction (PCR) and Western blotting (WB). Compared with rats in the control group, those in the model group had a disordered structure of hepatic lobule and hyperplasia of a large number of fibrous tissues. In contrast to those in the model group, the liver lobule structure was greatly improved, the edema cells were obviously reduced, and the hyperplasia of collagen fibers was remarkably alleviated in the probiotic group. Moreover, the degree of liver cirrhosis in the probiotic group was significantly reduced compared with that in the model group. Moreover, the rats in the model group exhibited a higher Bifidobacterium level in the intestinal tract, while those in the probiotic group displayed higher levels of microorganisms in the intestinal tract, such as Lachnospiraceae, Ruminococcaceae, Actinbacteria, Slackia and Pasteurellaceae. In comparison with that in the control group, the level of salt-tolerant Lactobacillus in the intestinal tract of rats in the model group was significantly decreased, while that in the probiotic group was partially increased (P=0.023). Meanwhile, some intestinal flora of rats in the control group, model group and probiotic group were closely correlated. Specifically, highly positive correlations were found between Bacteroidetes and Paraeggerthella (r=0.423, P=0.034) and between Firmicutes and Lactobacillus (r=0.318, P=0.027), but strongly negative associations were detected between Firmicutes and Paraeggerthella (r=-0.691, p=0.004) and between Paraeggerthella and Lactobacillus (r=-0.384, P=0.047). In addition, the levels of inflammatory cytokines TNF-α IL-6, MCP-1 and IFN-γ in the plasma of rats in the model group were markedly higher than those in the control group (P<0.05), whereas such levels in the probiotic group were decreased compared with those in the model group (P<0.05). PCR results revealed that the expression levels of β-catenin and Frizzled-2 in the model group were higher than those in the control group, whereas they were lower in the probiotic group than those in the model group (P<0.05). Furthermore, the model group had a decreased level of GSK-3β in comparison with the control group, but the probiotic group had a higher level of GSK-3β than the model group (P<0.05). WB results were consistent with PCR results. Probiotics can affect intestinal flora, inflammation and degree of liver cirrhosis in rats with liver cirrhosis, and its mechanism may be related to the Wnt/β-catenin signaling pathway.

MicroRNA-322/424 promotes liver fibrosis by regulating angiogenesis through targeting CUL2/HIF-1α pathway

AimsTo investigate the effects and mechanism of miR-322/424 in liver fibrosis. Main methodsmiR-322/424 expression in liver cirrhosis patients, mouse and rat liver fibrosis was determined by qPCR. Mice liver fibrosis was established by CCl4, and intervened by miR-322/424 agomir or antagomir. Liver hydroxyproline content and Sirius red staining were used to evaluate collagen deposition. CD31 expression was used to evaluate liver microvessel density. In vitro, the effects of miR-322/424 mimic or inhibitor on human hepatic sinusoidal endothelial cells (HHSECs) migration and tube formation were investigated. A dual luciferase reporter assay was performed to confirm the direct interaction between miR-322/424 and Cullin2. mRNA expression of elongin B/C, Cullin2, and RBX1 was determined by qPCR. HIF-1α protein expression was determined by Western blotting. Key findingsmiR-322/424 level in liver cirrhosis patients, mouse liver fibrosis induced by CCl4 and BDL, and rat liver fibrosis induced by CCl4 and dimethylnitrosamine was increased. miR-322/424 agomir exacerbated CCl4-induced mouse liver fibrosis, whereas the opposite effect was observed for miR-322/424 antagomir. miR-322/424 agomir significantly upregulated liver CD31 expression; opposite effects occurred with miR-322/424 antagomir. In vitro, miR-322/424 mimic significantly promoted tube formation and cell migration, and increased von Willebrand factor expression, whereas miR-322/424 inhibitor had the opposite effect. Dual-Luciferase Reporter Assay identified Cullin2 as miR-322/424 target. miR-322/424 decreased the mRNA expression of elongin B/C, Cullin2, and RBX1 and increased HIF-1α protein expression in HHSECs. SignificancemiR-322/424 plays a central role in the pathogenesis of liver fibrosis by targeting Cullin2, and enhancing HIF-1α-mediated hepatic angiogenesis.

Garcinia mangostana peel extracts exhibit hepatoprotective activity against thioacetamide-induced liver cirrhosis in rats

• Hepatoprotective effect of Garcinia mangostana is reported using experimental rats. • TAA induced liver cirrhosis assessed immuno/histochemically and biochemically. • Garcinia mangostana significantly down-regulate the PCNA, α-SMA and TGF-β1. • The activities of SOD and CAT considerably enhanced with decline in MDA level. • The serum level of ALP, ALT, AST, bilirubin and albumin was normal with p < 0.0001. The study in vivo evaluated the influence of ethanolic extract of G. mangostana peel on the histology, immunohistochemistry, and biochemistry of thioacetamide (TAA) induced liver cirrhosis in Sprague Drawly (SD) rats. The rats were distinctly administered with two doses of G. mangostana (250 mg/kg and 500 mg/kg) with TAA showed a significant reduction in liver index and hepatocyte proliferation with much lesser cell damage. These groups were significantly down-regulated the PCNA, α-SMA, and TGF-β1. The liver homogenate exhibited increased antioxidant enzymes (SOD and CAT) activities accompanied with decline in malondialdehyde (MDA) level. The serum level of bilirubin, total protein, albumin and liver enzymes (ALP, ALT, and AST) were restored to normal and were comparable to that normal control and silymarin with TAA treated groups. Thus normal microanatomy of hepatocytes, suppression of PCNA, α-SMA, TGF-β, improved antioxidant enzymes, reduced MDA with restoration of liver biomarkers demonstrates hepatoprotective influence of G. mangostana .

Mallotus furetianus extract protects against ethanol-induced liver injury via the activation of the cAMP-PKA pathway.

The protective effects of Mallotus furetianus extract (MF) on liver fibrosis induced with ethanol were examined using in vivo and in vitro model. MF treatment suppressed plasma alanine aminotransferase and aspartate aminotransferase activities in ethanol plus carbon tetrachloride (CCl4)‐induced cirrhosis rat model. MF also suppressed the increase in type l collagen and α‐smooth muscle actin expression in the livers of ethanol plus CCl4‐induced rat by the maintenance of intracellular glutathione levels. Furthermore, we evaluated the effect of MF on the alcohol‐induced activation of hepatic stellate cells (HSCs), which are responsible for the increased production and deposition of the extracellular matrix in liver injury. Here, we observed the enhancement of the intracellular reactive oxygen species (ROS) levels and the increase in type I collagen and a‐SMA expression in HSCs activated with ethanol. However, the enhanced ROS levels were suppressed with the treatments of MF or diphenyleneiodonium (DPI). Furthermore, the treatment of MF or DPI suppressed the increase in type I collagen and a‐SMA expression activated with ethanol. We also observed that the treatment of MF or LY194002 suppressed the increase in type I collagen expression in HSCs activated with ethanol, suggesting that ethanol induced type I collagen expression via the PI3K‐Akt signaling pathway. On the other hand, the suppression of the synthesis of type I collagen in ethanol and MF‐treated HSCs was inhibited by H‐89. From these results, MF may suppress the increase in the activity of NADPH oxidase in HSCs activated with ethanol through the cAMP‐PKA pathway.

Hepatoprotective effects and mechanisms of Ixeris denticulate water extract on liver cirrhosis in experimental rat

BackgroundTo explore the protective effect and mechanisms of Ixeris denticulate water extract (IDWE) in the development of liver cirrhosis in experimental rat.MethodsSixty rats were randomly divided into five groups: control group, model group and IDWE (2, 4 and 8 g/kg) treatment groups. Alanine transferase (ALT), aspartate transaminase (AST), albumin (ALB), tumor necrosis factor-alpha (TNF-α), Interleukin (IL)-6 and IL-8 in serum and superoxide dismutase (SOD), malondialdehyde (MDA) in liver tissue were evaluated, respectively. The liver index, liver morphology and liver histopathological analysis were detected as a supportive data. The liver protein expression of Bcl-2 and Bax were assessed by western blot, and NF-κB p65 protein expression was determined by immunohistochemistry analysis.ResultsThe result showed that a significantly decrease in the levels of serum AST, ALT and serum inflammatory factors TNF-α, IL-6 and IL-8 in IDWE-treated rats. The levels of serum ALB and SOD in liver tissue were markedly increased after IDWE treated, compared with model rats. Furthermore, IDWE-treated group also exhibited a down-regulated protein expression of NF-κB p65 and Bax, up-regulated Bcl-2 protein expression.ConclusionsIDWE could effectively alleviate the course of liver cirrhosis in rat model, which may be a potent hepatoprotective agent in clinical therapy in the future.

Transplantation of fetal liver tissue coated by ultra-purified alginate gel over liver improves hepatic function in the cirrhosis rat model

In this study, we used a new coating agent, that is, ultra-purified alginate gel (UPAL), for fetal liver tissue transplantation. This study aims to compare the effect of UPAL with the effect of other coating agents on improving the effect of fetal liver tissue transplantation in a liver cirrhosis rat model. Prior to the transplantation of wild-type ED14 fetal liver tissues, various coating agents were separately applied on the liver surface of rats with cirrhosis. Then, we compared the engraftment area, engraftment rate and liver function level of these rats. As a result, coating the liver surface of a cirrhosis rat with UPAL obtained the best effect in terms of engraftment area and engraftment rate of the transplanted liver tissue and in the recovery of liver function compared with control group. Therefore, UPAL coating may serve as a novel strategy for liver organoid transplantation.

Trehalose reduces bone loss in experimental biliary cirrhosis rats via ERK phosphorylation regulation by enhancing autophagosome formation.

Bone loss is a severe complication of primary biliary cirrhosis (PBC). Trehalose was intermittently administered in bile duct-ligated (BDL) male rats, a PBC-related osteoporosis model, for 4weeks to reduce osteoporosis. Femoral bones were assessed ex vivo by micro computed tomography (CT) and histomorphometry. The potential mechanisms related to the reduction of osteoporosis were explored by evaluating the effect of trehalose on osteoblast autophagy, osteogenesis, osteoclastogenesis, and ERK phosphorylation. The results demonstrated that trehalose reduced osteoporosis of BDL rats and decreased osteoblast-mediated osteoclast differentiation by enhancing osteoblast autophagy to regulate osteoprotegerin (OPG) secretion. Hydroxychloroquine (HCQ) increased the expression of OPG and OPG/receptor activator genes for nuclear factor-κB ligand (RANKL) ratio, and reduced osteoblast-mediated osteoclastogenesis by inhibiting autophagy flux and inducing autophagosome formation. Furthermore, trehalose increased the phosphorylation of ERK1/2 in MC3T3-E1 cells, and the ERK inhibitor PD98059 reversed the upregulation of OPG gene and reduction of trehalose-induced osteoclastogeneis. The treatment with HCQ markedly increased the ERK phosphorylation. The correlation between autophagosome formation and ERK phosphorylation was confirmed in autophagy proteins (ATG) 4B or ATG5-deficient cells. Thus, trehalose could decrease osteoblast-mediated osteoclastogenesis and reduce PBC-related bone loss by regulating ERK phosphorylation via autophagosome formation.

Effects of Thioacetamide—Induced Cirrhosis, after a Drug Washout Period, on the Michaelis–Menten Kinetics of Major Cytochrome P450 Enzymes in the Rat Liver Microsomes

PurposeThioacetamide (TAA) is used to induce liver cirrhosis in rodents. However, the drug itself directly inhibits cytochrome P450 (CYP450) enzymes. The purpose of this study was to determine the effects of TAA‐induced liver cirrhosis, after a washout period to eliminate TAA, on the Michaelis‐Menten (MM) kinetics and protein contents of the major CYP450 enzymes CYP2D, CYP2E1, and CYP3A in rat liver microsomes.MethodsAdult, male Sprague‐Dawley rats were treated for 10 weeks with either TAA (200 mg/kg ip, 3 days per week) or vehicle (n = 7/each group). After a 10‐day washout period, animals were euthanized, and the liver was collected after cardiac perfusion with cold saline. Liver microsomes (MC) were prepared by differential centrifugation. The activities of the liver microsomal CYP2D (dextromethorphan O‐demethylation), CYP2E1 (chlorzoxazone 6‐hydroxylation) and CYP3A (midazolam 1′‐hydroxylation) were estimated by measurement of the metabolites by LC‐MS/MS. The microsomal CYP450 reductase (CPR) activity was analyzed spectrophotometry. Furthermore, the protein expressions of the isoenzymes were determined using Western blot analysis. The MM kinetic parameters maximum velocity (Vmax) and MM constant (Km) were determined using nonlinear regression analysis. Unpaired t‐test was used for statistical analysis of data, with a p value of &lt; 0.05 as significant.ResultsWhereas the MM kinetics of dextromethorphan demethylation (CYP2D) were best fit by a two‐enzyme equation, those of chlorzoxazone hydroxylation (CYP2E1) and midazolam 1′‐hydroxylation (CYP3A) were best fit by a single‐enzyme equation. Cirrhosis resulted in significant reductions in the Vmax values for all the tested microsomal activities. The microsomal high‐ and low‐capacity Vmax values for CYP2D demethylase activity were significantly reduced by 74% (p value &lt; 0.0001) and 51% (p &lt; 0.05), respectively, in the TAA group. This was associated with a significant (p &lt; 0.001) reduction (71%) in the protein contents of CYP2D in the TAA group. However, Km values were not affected by cirrhosis. For CYP2E1, cirrhosis caused a significant reduction (52%) in the Vmax value (p &lt; 0.0001) for the chlorzoxazone hydroxylation activity. However, the Km values and protein contents of CYP2E1 remained unchanged. The Vmax and Km values of microsomal CYP3A hydroxylase activity, resulting in 1′‐hydroxylation of midazolam, were significantly reduced by 71% (p &lt; 0.001) and 46% (p &lt; 0.05), respectively, which was associated with a 30% reduction (p &lt; 0.05) in the protein contents of CYP3A. Cirrhotic animals also showed a 31% decline (p &lt; 0.05) in the microsomal CPR activity.ConclusionsTAA‐induced cirrhosis in rats resulted in a decreased liver microsomal activity of CPR and major CYP450 enzymes (CYP2D, CYP2E1, and CYP3A). Except for CYP2E1, the decreased isoenzyme activities were also associated with decreases in the protein concentrations of the isoenzymes. However, the cirrhosis‐induced decrease in the CYP2E1 activity, which was in the absence of a change in its protein levels, is possibly due to a decrease in the microsomal CPR activity.Support or Funding InformationFunding: Chapman University School of Pharmacy

Immunomodulatory effect of curcumin on hepatic cirrhosis in experimental rats.

Cirrhosis is a chronic liver disease. The present work aimed to evaluate the regulatory immune effect of curcumin in hepatic cirrhosis induced by carbon tetrachloride (CCl4) injections in experimental rats' model. Chronic liver fibrosis was induced in experiment animals by recurrent injections of CCl4 for more than 5weeks. They were divided into five groups: first group was injected with normal saline, second group with CCl4, third, fourth, and fifth groups were injected with CCl4 (intraperitoneal injection) at dose 3ml/kg, two times weekly for 6weeks supplemented with the administration of curcumin with concentrations 250, 200, and 150mg/kg. Immune response was analyzed to different treatments. Interleukin 10 (IL-10), pro-inflammatory cytokines TNF-α, TGF-1β, and liver histopathological examinations were conducted. The results showed that estimations of IL-10 concentrations were significantly increased in curcumin groups compared with CCl4 group, whereas TNF-α and TGF-1β levels were significantly decreased comparing with CCl4 group. The histopathological examinations for liver tissues showed that curcumin treated groups have almost retained the normal structure of liver tissues. In conclusion, curcumin inhibited hepatic fibrosis and liver fibrogenesis with regulation of the immune system mechanism against invader chemical toxicity. PRACTICAL APPLICATIONS: Curcumin is well documented for its medicinal properties, commonly used as a spice. Our work has thus demonstrated its effectiveness as an immunomodulatory agent. Practically, clinical studies have suggested that curcumin displays a diverse and powerful array of pharmacological effects in nearly all of the human body's major organ systems. These are: antidiabetes, anti-inflammatory, anticancer, antiaging, antioxidant, antibacterial infection, hepatoprotective, neurodegenerative, and cardiovascular effects.

Compromised Ileal Mucus Barrier Due to Impaired Epithelial Homeostasis Caused by Notch1 Signaling in Cirrhotic Rats.

In liver cirrhosis, intestinal mucus barrier is rarely studied. This study aimed to investigate whether mucus barrier in ileum is altered in cirrhotic rats and its underlying mechanisms. Thioacetamide was injected to induce liver cirrhosis in rats. Serum from portal vein blood, and ileum and liver tissues were obtained for further analysis. Goblet cell-like Ls174T cells were cultured for in vitro experiments. The ileal mucus was thin, loose, and porous with small bubbles in cirrhotic rats. mRNA expressions of Muc2 and TFF3 were also down-regulated in cirrhotic rats. Bacteria located near to crypts and LPS were increased in the serum from portal vein in cirrhotic rats. Smaller theca area and few goblet cells were found in cirrhotic rats compared with control. Increased proliferation of ileal epithelia was observed in cirrhotic rats. Notch1, Dll1, and Hes1 expressions were enhanced, and KLF4 expression was suppressed in ileum of cirrhotic rats. In Ls174T cells, EDTA and NICD plasmid induced NICD and Hes1 expression and suppressed KLF4 concomitantly, and mucus expression almost vanished in these cells. NICD plasmid induced more proliferation in Ls174T cells. Oppositely, after DBZ treatment, NICD and Hes1 were inhibited along with augmentation of KLF4 and increased mucous expression in Ls174T cells, while proliferation of the cells was suppressed. In cirrhotic rats, mucus barrier was impaired. This might be attributed to increased proliferation and decreased differentiation of epithelia, which might be mediated by Notch1-Hes1-KLF4 signaling.

A high-dose rapamycin treatment alleviates hepatopulmonary syndrome in cirrhotic rats.

Rapamycin is a type of immunosuppressive agent that acts through inhibition of mammalian target of rapamycin (mTOR). Hepatopulmonary syndrome (HPS) is a lethal complication in cirrhotic patients. It is characterized by hypoxia and increased intrapulmonary shunts, in which pulmonary inflammation and angiogenesis play important roles. The current study aimed to evaluate the effect of rapamycin on HPS using the experimental model of common bile duct ligation (CBDL)-induced cirrhosis in rats. The rats received low-dose (0.5 mg/kg), high-dose (2 mg/kg) rapamycin, or vehicle from the 15th to the 28th day post CBDL. Then the mortality rate, hemodynamics, biochemistry parameters, arterial blood gas and plasma levels of vascular endothelial growth factor (VEGF) and tumor necrosis factor (TNF)-α were evaluated on the 28th day post CBDL. Pulmonary histopathological stains were performed, and protein expression was examined. In parallel groups, the intrapulmonary shunts of CBDL rats were measured. Compared with the control, a high-dose rapamycin treatment decreased portal pressure and improved hypoxia in CBDL rats. It also reduced the plasma level of VEGF and TNF-α and decreased intrapulmonary shunts. Meanwhile, it ameliorated pulmonary inflammation and angiogenesis and downregulated the protein expression of mTOR, P70S6K, nuclear factor kappa B (NFκB), VEGF, and VEGF receptor 2. In contrast, low-dose rapamycin did not attenuate intrapulmonary shunts despite ameliorating portal hypertension. High-dose rapamycin ameliorates HPS in cirrhotic rats as evidenced by the alleviated hypoxia and decreased intrapulmonary shunts. Downregulation of the mTOR/P70S6K, NFκB, and VEGF signaling pathways might play a key role.

Vascular branching geometry relating to portal hypertension: a study of liver microvasculature in cirrhotic rats by X-ray phase-contrast computed tomography.

Portal hypertension is one of the major complications of cirrhosis. The changes in hepatic microvasculature are considered as critical pathophysiological characteristics of portal hypertension. X-ray phase-contrast computed tomography (PCCT) is a new imaging technique that can detect liver vessels at a micrometric level without contrast agents. In this study, male Sprague-Dawley rats with liver cirrhosis induced by carbon tetrachloride (CCl4) or bile duct ligation (BDL) were investigated with PCCT. The portal pressures of rats were recorded before euthanasia. The branch angle and Murray's deviation (MD) were measured based on the branching geometry of the three-dimensional (3D) microvasculature of liver cirrhosis in rats. Statistical analyses were performed to determine the correlation between branching geometry and portal pressure in liver fibrosis. The results demonstrated that the branch angle and MD significantly increased in the CCl4 model and BDL model compared with their corresponding normal group or sham group. The portal pressure was significantly correlated with the branching morphologic features (all R≥0.761 and P<0.01). The branch angle and MD could accurately distinguish portal pressure in cirrhotic rats, suggesting that branching geometric characteristics of the microvasculature may be a promising marker in the prognosis of portal hypertension in liver cirrhosis.

СОДЕРЖАНИЕ РЕТИНОЛА И α-ТОКОФЕРОЛА ПРИ ЭКСПЕРИМЕНТАЛЬНОМ ФИБРОЗЕ ПЕЧЕНИ У КРЫС

Background. The content of retinol and α-tocopherol in the human body affects the development and progression of chronic liver diseases and is associated with the functioning of perisinusoidal lipocytes (HSC) and the state of biological membranes. Objective – to evaluate the content of retinol and α-tocopherol in blood plasma and liver tissue in the dynamics of experimental liver fbrosis in rats. Material and methods. Modeling of liver fbrosis / cirrhosis was carried out on sexually mature male rats by intraperitoneal administration of thioacetamide (TAA) solution at a dose of 200 mg / kg every other day for 4 and 12 weeks. The control group of animals received an equal volume of saline. The concentration of α-tocopherol and retinol was determined by S.L. Taylor’s method. Results. In rat liver preparations, 4 weeks after administration of TAA solution to animals, signs of FII-III stage of fbrosis were observed. According to electron microscopy, HSCs were in a transitional state and acquired a more elongated shape; the number of lipid inclusions in their cytoplasm decreased. The administration of TAA for 12 weeks led to the formation of liver cirrhosis in rats, with characteristic macro- and microscopic changes. On light microscopy, the number of HSCs decreased in rat liver preparations 3 months after administration of TAA; activated HSCs were encountered, which acquired an elongated shape and lost lipid inclusions. The content of retinol in the 2nd group of animals (with liver fbrosis stage II-III) was 2.2 times higher than in the control group, and 1.8 times higher than in the 3rd group with liver cirrhosis (p &lt; 0.05). The content of retinol in the liver tissue after 4 weeks of TAA administration decreased by 11.7%, after 12 weeks - by 1.5 times. The level of α-tocopherol in the liver at the stage of fbrosis FII-III decreased by 21% compared with the control group, at the stage of cirrhosis - by 2 times. Conclusion. The use of thioacetamide in rats for 1 and 3 months leads to the development of liver fbrosis and cirrhosis. A decrease in the content of retinol and α-tocopherol in the liver occurs with the progression of liver fbrosis /cirrhosis. The high content of retinol and α-tocopherol in plasma at the stage of liver fbrosis FII-III is due to degranulation (activation) of HSC.

Bone marrow derived mesenchymal stem cells suppress diethylnitrosamine induced liver cirrhosis in rats

Aim: The present study was designed to explore the ability of bone marrow derived mesenchymal stem cells (BM-MSCs) to repair diethylnitrosamine (DEN)-induced liver cirrhosis (DILC) in rats. Materials and methods: Sixty male rats were divided into four groups (15 rats per group), the rats of the first group received saline, and the rats of the other three groups received DEN in drinking water. After thirty days, the rats of the third and fourth groups received 1.5x106 and 2x106 MSC infusions ,respectively. Liver function tests were estimated in serum of rats in all groups. The expression of albumin (Alb) and cytokeratin-18 (CK-18) genes were detected by reverse transcription-polymerase chain reaction (RT-PCR). Histopathological examination of liver tissue was performed. Results: Administration of BM-MSC of both doses (1.5x106 and 2x106 cells) into rats with DILC resulted in ameliorating liver functions and histopathological features compared to that received DEN only. RT-PCR analysis revealed that the expression of albumin gene increased, while CK-18 decreased in liver tissue of rats with DILC which received 2x106 BM-MSCs compared to DEN control group. Conclusion: BM-MSCs suppressed DEN-induced liver cirrhosis in rats as indicated by up regulation of albumin and CK-18 genes, as well as improvement of liver functions and architecture.

Impairment of natriuresis and diuresis induced by intrarenal adrenoceptor mechanisms in an experimental model of cirrhosis in rats

The contribution of intrarenal alpha-2 adrenergic receptors in mediating the enhanced renal excretory responses evoked by the alpha-2-agonist xylazine was examined in a model of cirrhosis in rats. In sham-operated rats, xylazine (0.2 mg/kg, i.v.) increased diuresis and natriuresis (urine flow, control: 78 ± 12.1, 10 min: 155 ± 17, 20 min: 194 ± 19, 30 min: 146 ± 16, 40 min: 114 ± 13, 50 min: 95 ± 10.5 μl/min/g; urinary sodium excretion, control: 6.75 ± 2.08, 10 min: 7.12 ± 2.1, 20 min: 13.4 ± 4.6, 30 min: 14.6 ± 4.02, 40 min: 12.05 ± 2.35, 50 min: 12.7 ± 2.45 μeq/min/g), which was accompanied by a significant reduction in renal sympathetic nerve activity (RSNA) (control: 100, 10 min: 39.5 ± 5.8, 20 min: 53 ± 8.8, 30 min: 72 ± 7.0, 40 min: 83 ± 5.0, 50 min: 94 ± 6.1 AU). Xylazine (0.2 mg/kg) in cirrhotic animals, despite resulting in a significant reduction in RSNA (control: 100, 10 min: 73 ± 4.3*, 20 min: 70 ± 5.0*, 30 min: 76 ± 7.0*, 40 min: 85 ± 5.5*, 50 min: 92 ± 4.8* AU), was unable to increase natriuresis. A higher dose (20 mg/kg) of xylazine was not capable of increasing natriuresis and diuresis, even in the presence of a robust reduction in RSNA. Renal denervation did not alter the onset and time course of cirrhosis. The results indicated that during the development of cirrhosis, there is an adaptive process that disables the intrarenal alpha-2 adrenoceptor mechanisms that selectively promote water and urinary sodium excretion via a sympathetic renal nerve-independent mechanism. Thus, in cirrhotic rats, the diuresis/natriuresis induced by xylazine is independent on RSNA. Intrarenal and/or hormonal changes are probably involved in the impairment of xylazine-induced diuresis/natriuresis in cirrhosis.