CCl4 Administration Research Articles

Hepatocurative Properties of Eucalyptus camaldulensis Stem Bark Extract

The research investigates the hepatocurative potentials of Eucalyptus camaldulensis solvents extract on CCl4 induced liver damage using albino rats, partial characterization of the most active extract was done using GCMS techniques. Forty-two rats were placed into seven groups of six rats each. Group I and II were normal control and test control respectively. Groups III to VII were induced with liver damage using 120mg/kg of CCl4 and administered with standard drug, n-hexane extract (E1), chloroform extract (E2), ethyl acetate extract (E) and methanol extract (E4) respectively. 24 hours after CCl4 administration, a significant (p<0.05) increase in mean serum level activities of AST, ALT, ALP with decrease in conjugated bilirubin and albumin concentration was recorded in test control group compared to normal control. This was reversed upon administration chloroform and methanol extract with former been most active. Antioxidant analysis reveals a significant (p<0.05) decrease in level of liver tissue antioxidant enzymes (SOD, CAT and GPx) with concomitant increase in MDA of test control compared to normal control rats. After extracts administration, increase in the antioxidants enzymes (SOD, Catalase and GPx) levels was recorded in methanol and standard drug administered groups.

Proanthocyanidins attenuated liver damage and suppressed fibrosis in CCl4-treated rats

Liver damage and fibrosis are serious health problems without effective treatment. Proanthocyanidins (PAs) are flavonoids with several biological effects. We investigated the potential anti-fibrotic effect of proanthocyanidins on carbon tetrachloride (CCl4)-induced liver injury and fibrosis. Liver fibrosis was induced by oral administration of CCl4 three times a week for 5 and 9 weeks. PAs were daily administered in a dose of 500 mg/kg bw. Animals were divided into five groups: control groups, olive oil-treated group, Pas-treated group, CCl4-treated animals, and PAs + CCl4-treated rats. CCl4 and PAs were administered by gavage. Administration of CCl4 caused a significant elevation in alanine aminotransferase and aspartate aminotransferase activities, the concentration of alpha-2-macroglobulin, and bilirubin concentration. In addition, the protein and apolipoprotein contents were significantly decreased in the serum of CCl4-treated rats. These results were accompanied by histopathological alterations and increased inflammation, apoptosis, and DNA damage. Treatment with PAs caused remarkable regression of fibrosis and alpha-2-macroglobulin with improvement in histological characteristics of the liver after 5 and 9 weeks of intoxication. PAs could also maintain redox balance, evidenced by the prevention of lipid peroxidation and mitigation of the decrease in antioxidants. Treatment of intoxicated rats with PAs resulted in a significant decline in pro-inflammatory cytokines, including IL-6, IL-1β, and TNF-α in serum. This is associated with a remarkable decrease in apoptosis of hepatic cells shown by decreased levels of Bax, caspase-3, and -9, with increased Bcl-2. The protective effect of PAs was also evident by protecting DNA integrity in the intoxicated rats. PAs suppressed hepatic fibrosis, improved liver function and structure via modulating the interdependence between oxidative stress, inflammation, apoptosis, and DNA integrity in CCl4-treated rats.

Lack of Nogo-B expression ameliorates PPARγ deficiency-aggravated liver fibrosis by regulating TLR4-NF-κB-TNF-α axis and macrophage polarization

Liver fibrosis is an important pathologic process in response to chronic or repetitive liver injury. It can advance to liver cirrhosis. Both peroxisome proliferator-activated receptor gamma (PPARγ) and Nogo-B play critical roles in fibrogenesis, while PPARγ is essential for the development. However, the effect of Nogo-B deficiency on the development of liver fibrosis in cell-specific PPARγ deficient mice remains unknown. In this study, hepatocyte or macrophage PPARγ deficient (hPPARγ KO or mPPARγ KO) mice, Nogo-B deficient mice, and cell-specific PPARγ plus Nogo-B double deficient (hPPARγ/Nogo-B DKO or mPPARγ/Nogo-B DKO) mice were induced liver fibrosis by CCl4 injection. We found hPPARγ KO mice showed enhanced liver fibrotic signatures compared to mPPARγ KO mice after CCl4 administration. Hepatocyte or macrophage PPARγ deficiency further enhanced CCl4-induced severe inflammation infiltration, apoptosis and M1 macrophage polarization in the liver. In contrast, Nogo-B deficiency effectively ameliorated PPARγ deficiency-aggravated liver injury and fibrosis. It ameliorated PPARγ deficiency-aggravated liver inflammation and fibrosis by suppressing hepatic stellate cell activation, TLR4-NF-κB/TNF-α signaling and M1 macrophage polarization. In conclusion, our study demonstrates that PPARγ deficiency increases susceptibility of mice to develop CCl4-induced liver injury/fibrosis, which is potently reduced by Nogo-B deficiency, indicating Nogo-B inhibition might be a therapeutic approach for liver fibrosis treatment.

Liposomal Drug Delivery of Blumea lacera Leaf Extract: In-Vivo Hepatoprotective Effects.

Background: Blumea lacera (B. lacera) is a herbaceous plant commonly found in south-east Asia. It shows significant therapeutic activities against various diseases. The objectives of this study were to evaluate hepatoprotective effects of Blumea lacera leaf extract and also to investigate the comparative effectiveness between a liposomal preparation and a suspension of B. lacera leaf extract against carbon tetrachloride (CCl4)-induced liver damage. Methods: B. lacera leaf extract was characterized using a GC-MS method. A liposomal preparation of B. lacera leaf extract was developed using an ethanol injection method and characterized using dynamic light scattering (DLS) and electronic microscopic systems. The hepatoprotective effects of B. lacera leaf extracts and its liposomal preparation were investigated using CCl4-induced liver damage in Long Evan rats. Results: GC-MS data showed the presence of different components (e.g., phytol) in the B. lacera leaf extract. DLS and microscopic data showed that a liposomal preparation of B. lacera leaf extracts was in the nano size range. In vivo study results showed that liposomal preparation and a suspension of B. lacera leaf extract normalized liver biochemical parameters, enzymes and oxidative stress markers which were elevated due to CCl4 administration. However, a liposomal formulation of B. lacera leaf extract showed significantly better hepatoprotective effects compared to a suspension of leaf extract. In addition, histopathological evaluation showed that B. lacera leaf extract and its liposomal preparation treatments decreased the extent of CCl4-induced liver inflammations. Conclusion: Results demonstrated that B. lacera leaf extract was effective against CCl4-induced liver injury possibly due to the presence of components such as phytol. A liposomal preparation exhibited significantly better activity compared to a B. lacera suspension, probably due to improved bioavailability and stability of the leaf extract.

Protective effect of acrocarpus fraxinifolius extract against hepatic fibrosis induced by Gamma irradiation and carbon tetrachloride in albino rats

Purpose Liver fibrosis is considered as one of the ultimate outcomes of chronic liver disorders, characterized by outrageous cell proliferation and abnormal deposition of extracellular matrix, resulting in sever pathological distortions in the architecture and performance of liver tissues. The present study aimed to investigate the protective properties of aqueous methanol extract of Acrocarpus fraxinifolius leaves (AFL) against liver fibrosis induced by dual toxicity of γ-irradiation and carbon tetrachloride (CCl4) in rats. Methods The animals were exposed to 2 Gy irradiation once/week concurrently with intraperitoneal administration of CCl4 (0.2 mL/100 g body weight) for seven weeks. Afterwards, liver toxicity and fibrosis were assessed biochemically at cellular and molecular as well as histopathological levels. Results The livers of intoxicated rats showed distinct structural and functional changes, compared with the normal rats. The administration of AFL (500 mg/kg, p.o) significantly ameliorated the histopathological manifestations of fibrotic liver evidenced by mitigated steatosis progression, necrosis, fibrotic septa, apoptotic bodies, and immunochistochemical studies of alpha-smooth muscle actin. Also, AFL increased the final body weight, total protein, albumin levels and albumin/globulin ratio. While, the absolute liver weight, liver enzymes, total cholesterol and triglycerides were reduced. A significant modulation was observed in hydroxyproline, transforming growth factor-β and collagen-1expression. Furthermore, AFL exerted a direct effect on liver fibrosis by promoting extracellular matrix degradation via overexpression of the tissue inhibitor metalloproteinase-1, coupled with decease of metalloproteinase-9 activity. Conclusions Our findings suggested that AFL effectively improved the architecture of fibrotic liver and modified the biochemical markers of liver fibrosis.

Nitric oxide-mediated effects of L-ornithine-L-aspartate in acute toxic liver injury

This study was aimed to investigate nitric oxide-dependent mechanisms of L-ornithine-L-aspartate (LOLA) action in acute toxic liver injury in rats. Acute hepatitis was induced in Wistar rats using 50% oil solution of tetrachloromethane (CCl4) intragastrically (2 g/kg) twice in a 24 hour interval. Intraperitoneal treatment with LOLA (200 mg/kg) was started 6 hours after the second CCl4 administration and maintained for 3 consecutive days. L-Nω-Nitroarginine Methyl Ester (L-NAME) was used intraperitoneally (10 mg/kg). In CCl4-induced hepatitis, LOLA restores the structure of hepatocytes and prevents aminotransferases, alkaline phosphatase and gamma-glutamyl transferase elevation. It decreases total bilirubin concentration but does not affect increased cholesterol level. LOLA augments urea concentration, total protein level in blood and liver as well as serum and liver content of nitrite anions. LOLA enhances activity of catalase, glutathione S-transferase, manganese superoxide dismutase, increases reduced glutathione level and total antioxidant capacity and decreases thiobarbituric acid reactive substances level. The concomitant use of L-NAME inhibits the action of LOLA to enhance nitrite anions synthesis both in serum and liver, to delay the recovery of hepatocytes, to counteract LOLA effect against blood total protein reduction, to prevent the decline in aminotransferases, alkaline phosphatase,, gamma-glutamyl transferase and glutathione S-transferase activity and to reduce catalase activity and reduced glutathione level. Therefore, in CCl4-induced hepatitis, LOLA effectively prevents cytolysis and cholestasis, improves liver metabolism and protects against oxidative stress. Partially, these changes occur in nitric oxide-mediated mechanism since the use of L-NAME declines most of LOLA effects.

The therapeutic potential of Nelumbo Nucifera against oxidative damage induced by Carbon Tetrachloride in rats.

Background: Herbal treatment has paid much attention in the world over the past decades. Nelumbo nucifera is used all over the world as a medicinal plant. It is an aquatic perennial plant belonging to the Nelumbonaceae family. The protective effects of Nelumbo nucifera are due to the active ingredients present in it. The study was conducted to evaluate the protective effects of Nelumbo nucifera seeds against hepatotoxicity caused by carbon tetrachloride (CCl4) in rats. Methodology: Eighteen Wistar Albino rats were allotted into three groups (n=6), control group, CCl4 treated group given CCl4 0.8 ml/kg body weight subcutaneously twice a week, CCl4 + N. Nucifera treated group given CCl4 0.8 ml/kg body weight 2 times a week subcutaneously and also given Nelumbo nucifera extract (200 mg/kg body weight) orally through gavage daily. After 28 days of the experiment, serum samples were taken for estimation of enzyme activities and liver tissue samples were collected for evaluation of antioxidant enzyme levels. Results: It was observed that liver weight increased in the CCl4 treated group while liver weight decreased in CCl4 + N. Nucifera treated group. CCl4 administration significantly raises the transaminase levels such as Aspartate transaminase (AST) (p<0.01), Aspartate transaminase (ALT) (p<0.05), and Alkaline phosphatase (ALP) (p<0.05) whereas CCl4 + N. Nucifera treated group serum transaminases are reduced. Antioxidant activities such as Catalase (p>0.05), Glutathione (GSH) (p>0.05) and Superoxide dismutase (SOD) (p=0.05) decreased in the CCl4 treated group. Significant elevation of antioxidant enzymes; Catalase (p<0.05), GSH (p<0.05) and SOD (p>0.05) in CCl4 + N. Nucifera treated group. Serum Malondialdehyde (MDA) concentration (p<0.05) significantly raised in the CCl4 hepatotoxic group compared to the control group. A decreased MDA levels were observed in the CCl4 + N. Nucifera treated group, indicating its potential to counteract the harmful effects of CCl4. Conclusion: This study suggests that Nelumbo nucifera seeds have overcome CCl4-induced hepatotoxicity by their antioxidant effects.

D-Carvone Attenuates CCl4-Induced Liver Fibrosis in Rats by Inhibiting Oxidative Stress and TGF-ß 1/SMAD3 Signaling Pathway.

Simple SummaryLiver fibrosis is a challenging global health problem resulting in a significant morbidity and mortality rates worldwide due to its rapid progression to cirrhosis and hepatocellular carcinoma. Therefore, identifying nontoxic therapies with precise curative effects to slow the progression of liver fibrosis comprises one of the most popular and high-priority areas of current research. D-carvone is a naturally occurring monoterpene abundant in the essential oil of aromatic plants such as caraway and spearmint. In the present study, the protective impact of D-carvone on carbon tetrachloride (CCl4)-induced liver fibrosis in rats was evaluated. Administration of D-carvone significantly enhanced liver functions, oxidant/antioxidant balance as well as liver histology. D-carvone ameliorated the progression of liver fibrosis, evident by the decreased collagen deposition (fibrosis score) and the reduced expression of the pro-fibrogenic markers TGF-β1 and SMAD3 in the liver. These findings reveal the anti-fibrotic effects of D-carvone and suggest that D-carvone could be a promising candidate for therapeutic intervention of liver fibrosis and other oxidative stress-related hepatic diseases.D-carvone is a natural monoterpene found in abundance in the essential oil of aromatic medicinal plants with a wide range of pharmacological values. However, the impact of D-carvone on liver fibrosis remains unclear. This study aimed to evaluate the anti-fibrotic potential of D-carvone in a rat model of liver fibrosis and to clarify the possible underlying mechanisms. Liver fibrosis was induced in rats by carbon tetrachloride, CCl4 (2.5 mL/kg, interperitoneally every 72 h for 8 weeks). Oral treatment of rats with D-carvone (50 mg/kg, daily) started on the 3rd week of CCl4 administration. D-carvone significantly enhanced liver functions (ALT, AST), oxidant/antioxidant status (MDA, SOD, GSH, total antioxidant capacity; TAC), as well as histopathological changes. Moreover, D-carvone effectively attenuated the progression of liver fibrosis, evident by the decreased collagen deposition and fibrosis score by Masson trichrome staining (MT) and α-SMA protein expression. Moreover, D-carvone administration resulted in a significant downregulation of the pro-fibrogenic markers TGF-β1 and SMAD3 and upregulation of MMP9. These findings reveal the anti-fibrotic effect of D-carvone and suggest regulation of the TGF-β1/SMAD3 pathway, together with the antioxidant activity as a mechanistic cassette, underlines this effect. Therefore, D-carvone could be a viable candidate for inhibiting liver fibrosis and other oxidative stress-related hepatic diseases. Clinical studies to support our hypothesis are warranted.

D-limonene (5 (one-methyl-four-[1-methylethenyl]) cyclohexane) diminishes CCl4-induced cardiac toxicity by alleviating oxidative stress, inflammatory and cardiac markers

ABSTRACT Background: The cardiovascular crisis is advancing rapidly throughout the world. A large number of studies have shown that plant polyphenols affect major mechanisms involved in cardiovascular events through their action on the antioxidant system, signaling, and transcription pathways. D-limonene, a monocyclic monoterpene obtained from citrus fruits, is reported to possess many pharmacological activities. Methods: The experiment was designed to determine the protective effect of D-limonene against cardiac injury induced by CCl4 in Wistar rats. Rats were treated with two doses of D-limonene against cardiac injury induced by CCl4. Serum toxicity markers, cardiac toxicity biomarker enzymes, inflammatory mediators, anti-oxidant armory, lipid peroxidation, lipid profile, and histology were done. Results: CCl4 intoxication resulted in a substantial rise in FFA, TC, TG, PL, LDL, VLDL, and a reduction in HDL, restoring these changes with the administration of D-limonene at a dosage of 200 mg/kg. CCl4 administration also resulted in lipid oxidation and decreased antioxidant activity. At the same time, D-limonene at a dosage of 200 mg/kg body weight inhibited LPO and restored in vivo antioxidant components to normal. CCl 4 intoxication also resulted in a significant increase in inflammatory markers like IL-6, TNF-α, high sensitivity Corticotropin Releasing Factor (Hs-CRF), and biomarkers of cardiac toxicity like alanine aminotransferase (ALT), lactate dehydrogenase (LDH), creatine kinase (CK), creatine kinase MB (CKMB), and Troponin I & troponin-t activities. D-limonene reversed all these changes to normal. Histology further confirmed our obtained results. Conclusion: These findings indicate that D-limonene can ameliorate cardiac injury at a 200 mg/kg body weight dosage. Henceforth, D-Limonene intervenes in mediating CCl4 induced toxicity by various signaling pathways.

Evaluation of hepatoprotective effect of Withania somnifera and Rubia cordifolia against carbon tetrachloride induced hepatotoxicity in albino rats

Liver is one of the largest gland of the body and the main site for intense metabolism and excretion. Liver is concerned with metabolism of endogenous as well as exogenous substances. “Withania somnifera” and “Rubia cordifolia” claim to have hepatoprotective action. Till now, no scientific study has been performed to evaluate such claims. This was an experiment study conducted on wistar rats, in Department of Pharmacology, Index Medical College. Healthy Albino wistar rats of either gender, weighing 150- 200g were obtained from CPCSEA approved Central Animal House. The selected rats were housed in polpropylene cages under controlled conditions of temperature (25 0C) and alternating periods of light and darkness of 12 hours each. The rats had free access to standard rat pellet diet and tap water ad libitum. After one week of acclimatization, the animals were rendered suitable for study. Pregnant female rats were not included in the study. ALT level in normal saline treated group was 29.5±3.35 IU/L. It was found to be significantly increased (p<0.001) with administration of CCl4 to 433.5±48.67 IU/L.

Invivo partial cellular reprogramming enhances liver plasticity and regeneration.

Mammals have limited regenerative capacity, whereas some vertebrates, like fish and salamanders, are able to regenerate their organs efficiently. The regeneration in these species depends on cell dedifferentiation followed by proliferation. We generate a mouse model that enables the inducible expression of the four Yamanaka factors (Oct-3/4, Sox2, Klf4, and c-Myc, or 4F) specifically in hepatocytes. Transient invivo 4F expression induces partial reprogramming of adult hepatocytes to a progenitor state and concomitantly increases cell proliferation. This is indicated by reduced expression of differentiated hepatic-lineage markers, an increase in markers of proliferation and chromatin modifiers, global changes in DNA accessibility, and an acquisition of liver stem and progenitor cell markers. Functionally, short-term expression of 4F enhances liver regenerative capacity through topoisomerase2-mediated partial reprogramming. Our results reveal that liver-specific 4F expression invivo induces cellular plasticity and counteracts liver failure, suggesting that partial reprogramming may represent an avenue for enhancing tissue regeneration.

Histomorphometric effects of Raphanus Sativus leaf extract on Carbon Tetrachloride induced hepatotoxicity

Background: Raphanus sativus is a cruciferous plant which possess free radical scavenging properties that defend the body against the deleterious effects of reactive oxygen species. To evaluate the protective role of Raphanus sativus in CCl4-induced hepatotoxicity through assessment of liver function and oxidative stress markers in albino Wistar rats.
 Material and Methods: Quasi-experimental study was conducted from September 2019 to March 2020 at the Postgraduate Research Laboratory, Isra University, Hyderabad. Non-random purposive sampling was used for selecting Albino Wistar rats. All rats were then divided equally into three different groups: Group A (control group), Group B (CCl4 experimental group), and Group C (CCl4 plus Raphanus sativus group). Blood samples were collected through cardiac puncture followed by hepatic histopathological analysis using light microscope. Data was analyzed using SPSS version 24, with ANOVA and Post hoc Tukey analysis used for comparison of different study variables.
 Results: Statistically significant rise relative liver weight of group B rats compared with the group A and C rats (P <.05). Moreover, statistically significant (P <0.05) rise in serum markers of hepatic functions was observed after CCl4 administration in Group B. Treatment with Raphanus Sativus administration significantly reduced serum levels of LFTs (p<0.05). There was a significant decline in the plasma levels of oxidative markers in group B while marked histopathological changes like necrosis, sinusoidal dilatation and congestion observed among animals of group B.
 Conclusion: Raphanus Sativus exerts an anti-oxidative, anti- and hepatoprotective effect against CCl4-induced hepatic tissue damage.

Protective effect of Sophora pachycarpa seed extract on carbon tetrachloride-induced toxicity in rats

The aim of the present study was to investigate the protective effect of the Sophora pachycarpa (S. pachycarpa) seed extract against carbon tetrachloride-induced toxicity on body organs, blood, and biochemical factors. In this investigation, 40 male Wistar rats weighing 200–250 g were randomly divided into 5 groups: group I was used as control, group II received carbon tetrachloride (CCl4) (IP, 1 mL/kg) on day 21, group III and group IV received S. pachycarpa seed extract at doses of 150 mg/kg and 300 mg/kg, respectively for 21 days by oral gavage and CCl4 on day 21, group V received silymarin (300 mg/kg) for 21 days by oral gavage and CCl4 on day 21. CCl4 showed an increase of serum renal and hepatic markers creatinine, urea, blood urea nitrogen (BUN), and uric acid, alkaline phosphatase (ALP), aspartate aminotransferase (AST), and alanine aminotransferase (ALT). Also, it significantly increased MDA level, and decreased CAT, FRAP, GSH, and SOD in the liver and kidney tissues. These changes and also hematological and histopathological alterations were significantly ameliorated by S. pachycarpa seed extract before CCl4 administration. In conclusion, the data obtained in our investigation confirm the protective effect of S. pachycarpa against acute exposure to CCl4-induced organ toxicity in rats.

Evaluation of Hepatoprotective Potential of Polyherbal Preparations in CCl4-Induced Hepatotoxicity in Mice.

Background Polyherbal formulations (PLFs) have been widely used for liver protection, treatment for hepatic dysfunction, and regeneration. They can also enhance appetite and protect the gastrointestinal tract from injury. In spite of the prevalent use, there is a need of scientific evidence on their effectiveness and safety. The objective of the present study was to assess the hepatoprotective effect of polyherbal formulations (commercially available in Bangladesh namely Heptaliv, Holyliv, Icturn, and J-deenar) in CCl4-induced hepatotoxicity in mice. Methods In this study, Swiss albino mice were treated for 7 days with distilled water or PLFs (2.6 and 5.2 ml/kg body weight/day, per os.) followed by single subcutaneous injection of CCl4 (1 ml/kg body weight, diluted with olive oil in 1 : 1 ratio) on day 8. Twenty-four hours after CCl4 administration, the mice were monitored for the effects of PLFs on liver morphology, biochemical parameters including serum aspartate transaminase (AST), serum alanine transaminase (ALT), alkaline phosphatase (ALP), and total bilirubin. Phenobarbitone-induced sleeping time and histopathology changes in liver tissues were also monitored. Results CCl4 administration caused significant hepatotoxicity as evidenced by marked elevation in AST, ALT, ALP, and total bilirubin. Phenobarbitone-induced sleeping time and infiltration of inflammatory cells and centrizonal necrosis on histological examination of liver demonstrated hepatic injury after CCl4 administration. However, the administration of Icturn and J-deenar polyherbal formulations at the higher dose significantly decreased the levels of AST, ALT, ALP, and total bilirubin. Moreover, pentobarbitone-induced sleeping time and histopathological analysis also revealed significant improvement as result of treatment with formulations Icturn and J-deenar. Conclusion Our results confirmed that polyherbal formulations (Icturn and J-deenar) can significantly prevent CCl4-induced hepatotoxicity in mice, demonstrating their protective effect for liver.

Methanol Stem Bark Extract of Ficus platyphylla Protects Against Carbon Tetrachloride-Induced Liver Damage in Wistar Rats

Ficus platyphylla (Gutta percha tree) is used in Nigerian traditional medicine for treating pain and psychosis. This study evaluated the protective effect of methanol stem bark extract of Ficus platyhylla (MEFP) against carbon tetrachloride (CCl4)-induced liver injury in Wistar rats. Rats were allotted into six groups namely normal control, negative control, standard drug (silymarin) and extract treated (100, 200 and 400 mg/kg p.o. respectively) groups. Treatment was administered for 7 days. Animals in groups II-VI received a single dose of CCl4 (1:1) p.o. in olive oil at 1.5 ml/kg body weight after last treatment. Experimental rats were sacrificed after 24 hours of CCl4 administration and blood samples collected for determination of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities. Supernatant of liver homogenate was used to determine activities of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx). Results indicate increased activities (p < 0.05) in CAT, SOD and GPx in samples of MEFP treated rats while levels of ALT and AST in serum samples of treated groups differ significantly (p<0.05) when compared with negative control group (Group III). Treatment with methanol stem bark extract of Ficus platyphylla may have offered protection against CCl4 induced liver damage in experimental rats.

Adaptive Mechanisms of Renal Bile Acid Transporters in a Rat Model of Carbon Tetrachloride-Induced Liver Cirrhosis.

Background: Acute kidney injury (AKI) is common in advanced liver cirrhosis, a consequence of reduced kidney perfusion due to splanchnic arterial vasodilation and intrarenal vasoconstriction. It clinically manifests as hepatorenal syndrome type 1, type 2, or as acute tubular necrosis. Beyond hemodynamic factors, an additional mechanism may be hypothesized to explain the renal dysfunction during liver cirrhosis. Recent evidence suggest that such mechanisms may be closely related to obstructive jaundice. Methods: Given the not completely elucidated role of bile acids in kidney tissue damage, this study developed a rat model of AKI with liver cirrhosis induction by carbon tetrachloride (CCl4) inhalation for 12 weeks. Histological analyses of renal and liver biopsies were performed at sacrifice. Organic anion tubular transporter distribution and apoptosis in kidney cells were analyzed by immunohistochemistry. Circulating and urinary markers of inflammation and tubular injury were assayed in 21 treated rats over time (1, 2, 4, 8, and 12 weeks of CCl4 administration) and 5 controls. Results: No renal histopathological alterations were found at sacrifice. Comparing treated rats with controls, organic anion transporters were differentially expressed and localized. High serum bile acid values were detected in cirrhotic animals, while caspase-3 staining was negative in both groups. Increased levels of serum inflammatory and urinary tubular injury biomarkers were observed during cirrhosis progression, with a peak after 4 and 8 weeks of treatment. Conclusions: These findings suggest possible adaptive tubular mechanisms for bile acid transporters in response to cirrhosis-induced AKI.

Evaluation of liver T1rho and T2 values in acute liver inflammation models using 7T-MRI

PurposeWe measured the T1rho and T2 values the liver of acute liver inflammation model mice administered carbon tetrachloride (CCl4) after 3 days and 6 days after dispensed, and we compared and examined whether each relaxation time can be used for detect acute liver inflammation. MethodsTo create an acute liver inflammation model, a mixture of 0.2 ml / 100 g of CCl4 with an equal amount of Sesame Oil was administered once intraperitoneally to C57BL / 6JJmsSlc mice (n = 15). On the 3 days and 6 days after administration, we acquired T1rho mapping images and T2 mapping images of the liver under respiratory synchronization using for preclinical 7T-MRI, and we measured T1rho and T2 values and compared statistically. ResultsThe liver T1rho value of control mice was 33.9 ± 2.5 ms before CCl4 administration, 43.2 ± 4.9 ms (p < 0.01) on the 3 days post CCl4 injection, and 41.0 ± 1.2 ms (p < 0.001) on the 6 days post CCl4 injection. The rate showed a significant increase of 27% on the 3 days after, as well as significant increase of 21% on the 6 days after. On the other hand, the liver T2 value of control mice was 26.7 ± 1.9 ms before CCl4 administration, 31.5 ± 3.4 ms (p < 0.05) 3 days post CCl4 injection, and 29.0 ± 2.0 ms (p = 0.06) 6 days post CCl4 injection. The rate 3 days after CCl4 administration showed a significant increase of 18%, after 6 days rate increased 9%, but no significant difference was confirmed compared with normal mice. ConclusionsThe T1rho value changed significantly compared to the T2 value, and a continuous change was observed even after 6 days. T1rho mapping can diagnose acute liver inflammation.

Tetramethylpyrazine prevents liver fibrotic injury in mice by targeting hepatocyte-derived and mitochondrial DNA-enriched extracellular vesicles.

Liver fibrosis is the common consequence of almost all liver diseases and has become an urgent clinical problem without efficient therapies. Recent evidence has shown that hepatocytes-derived extracellular vesicles (EVs) play important roles in liver pathophysiology, but little is known about the role of damaged hepatocytes-derived EVs in hepatic stellate cell (HSC) activation and following fibrosis. Tetramethylpyrazine (TMP) from Ligusticum wallichii Franchat exhibits a broad spectrum of biological activities including liver protection. In this study, we investigated whether TMP exerted liver-protective action through regulating EV-dependent intercellular communication between hepatocytes and HSCs. Chronic liver injury was induced in mice by CCl4 (1.6 mg/kg, i.g.) twice a week for 8 weeks. In the last 4 weeks of CCl4 administration, mice were given TMP (40, 80, 160 mg·kg-1·d-1, i.g.). Acute liver injury was induced in mice by injection of a single dose of CCl4 (0.8 mg/kg, i.p.). After injection, mice were treated with TMP (80 mg/kg) every 24 h. We showed that TMP treatment dramatically ameliorated CCl4-induced oxidative stress and hepatic inflammation as well as acute or chronic liver fibrosis. In cultured mouse primary hepatocytes (MPHs), treatment with CCl4 or acetaminophen resulted in mitochondrial dysfunction, release of mitochondrial DNA (mtDNA) from injured hepatocytes to adjacent hepatocytes and HSCs through EVs, mediating hepatocyte damage and fibrogenic responses in activated HSCs; pretreatment of MPHs with TMP (25 μM) prevented all these pathological effects. Transplanted serum EVs from TMP-treated mice prevented both initiation and progression of liver fibrosis caused by CCl4. Taken together, this study unravels the complex mechanisms underlying the protective effects of TMP against mtDNA-containing EV-mediated hepatocyte injury and HSC activation during liver injury, and provides critical evidence inspiring the development of TMP-based innovative therapeutic agents for the treatment of liver fibrosis.

Involvement of CD4+ and CD8+ T-lymphocytes in the modulation of nociceptive processing evoked by CCL4 in mice

AimsTo explore the mechanisms involved in the transformation of analgesia produced by low doses of CCL4 (pg/kg) to hyperalgesia when higher doses (ng/kg) are administered to mice. Main methodsThe unilateral hot plate test was used to assess thermal nociception. CD3+, CD4+ or CD8+ blood cells were depleted with selective antibodies. Expression of CCR5 and IL-16 in lymphocytes was studied by flow cytometry and IL-16 blood levels were measured by ELISA. IL-16 and CD8 were detected by immunofluorescence. Key findingsIL-16 and CCR5 expression were demonstrated in CD4+ and CD8+ T-lymphocytes by flow cytometry. Furthermore, CCL4-induced hyperalgesia was abolished by reducing circulating T-lymphocyte levels or by selectively depleting CD4+ lymphocytes. In contrast, when the anti-CD4 antibody was acutely administered, CCL4 induced analgesia instead of hyperalgesia. A similar response was obtained when administering A-770041, that prevents CD4-mediated CCR5 desensitization by inhibiting p56lck kinase. As occurred with the analgesic effect evoked by low doses of CCL4, analgesia evoked by combining CCL4 and A-770041 was reverted by naloxone, naltrindole or an anti-met-enk antibody. Interestingly, flow cytometry assays showed that the number of CD8+, but not CD4+, T-cells expressing IL-16 is reduced after the acute administration of CCL4, a result compatible with the description that CD8+-lymphocytes can rapidly release preformed IL-16. Accordingly, the rise in IL-16 blood concentration evoked by CCL4 was prevented after CD8+ lymphocyte depletion. SignificanceCCL4-evoked hyperalgesia is related to the desensitization of CCR5 in CD4+ T-cells and to the release of IL-16 from CD8+ lymphocytes.

HEPATOPROTECTIVE EFFECT OF CRYOPRESERVED PLACENTA EXTRACT ON THE TETRACHLOROMETHANE LIVER INJURY

Introduction. Toxic lesions of the hepatobiliary system are an important medical and social problem due to the steady increase in the frequency of chemical hepatosis, which develops as a result of accumulation of various xenobiotics in the body. Tetrachloromethane (CCl4) is one of the most studied hepatotoxins. Biochemical and pathomorphological changes in the liver lesions modeled by the administration of CCl4 are analogous to acute hepatitis of various etiologies in humans. As a potential hepatoprotective agent, cryopreserved placenta extract (CPE) attracted our attention. The objective is to characterize the effect of cryopreserved placenta extract on the functional state of the liver of rats with tetrachloromethane-induced damage. Materials and Methods. In vivo study was conducted on 28 male rats weighing 200–220 g. Acute CCl4-induced hepatitis was reproduced by injecting a 50.0% oil solution of CCl4 at a dose of 10 ml/kg body weight of the animal once, which caused acute fatty liver dystrophy. Animals were sacrificed 24 hours after the administration of CCl4. CPE was administered once per day for five days before CCl4 administration. The research material was blood and liver homogenates. The concentration of bilirubin in blood was determined by the spectrophotometrical method. The content of adenosine monophosphoric, adenosine diphosphoric, and adenosine triphosphoric acids was studied in the deproteinized homogenate by chromatographic method. Results and discussion. It was established that the development of CCl4-induced hepatitis is accompanied by the formation of a cholestatic syndrome and a disorder of energy metabolism in liver tissues. This was indicated by an increase (p &lt; 0.001) in the level of total bilirubin by 4.7 times and a decrease (p &lt; 0.001) in the energy charge by 42.6% compared to the indicators of intact rats. Prophylactic five-day administration of CPE led to a decrease (p &lt; 0.001) in the level of total bilirubin by 32.1% and a decrease (p &lt; 0.001) in the levels of direct and indirect bilirubin by 10.6% and 65.1%, respectively, compared to the indicators of animals with experimental toxic hepatitis. This indicated the weakening of the cholestatic syndrome and, indirectly, the weakening of inflammatory infiltration against the background of cytolytic processes in the liver. CPE had an energy-stabilizing effect on hepatocytes of rats with simulated tetrachloromethane liver injury. This was indicated by an increase (p = 0.02) of the energy charge by 18.2% compared to the indicators in the control group. Conclusions. The study showed anticholestatic and energy-stabilizing activity of the cryoextract of placenta used as a prophylactic measure in the model of tetrachloromethane-induced hepatitis.