CCl4 Solution Research Articles

Mechanism of action involved in the hepatoprotective activities of methanol extract of Cassytha filiformis L. aerial parts in CCl4-induced liver damage

Cassytha filiformis alcohol decoction is used by traditional healers in Idem-Ogwa, Mbaitoli Local Government Area of Imo State, Nigeria, in the treatment of liver disorders ranging from hepatitis to susceptive poisoning and alcohol intoxication. The methanol extract of the aerial parts of Cassytha filiformis L. was investigated for hepatoprotective activity in rats challenged with carbon tetrachloride (CCl4). Apart from group 1 (normal), all other groups were pre-challenged with a single intraperitoneal injection of 0.7 ml of 20 % (v/v) CCl4 solution in olive oil. Group 2 (negative control) was given distilled water (10 ml kg−1, p.o.). Group 3 were treated with silymarin (35 mg kg−1 p.o.). Groups 4, 5, and 6 received oral treatments of 75, 150, and 300 mg kg−1 of the extract of C. filiformis respectively. All treatments were given by stomach intubation and at 12 h intervals for 7 days. Sodium pentobarbitone-induced sleeping time assay was carried out on the animals at 12 h after the last treatment on day 7 by intraperitoneal administration of Na-pentobarbitone (35 mg kg−1). Serum from individual animal was used to determine the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total bilirubin, total protein and in vitro red blood cell membrane stability. The extract was safe up to 2000 mg kg−1 orally and did not induce overt clinical manifestation or death in rats. The extract significantly (p < 0.01) demonstrated potency at all test doses (75, 150, and 300 mg kg−1) in CCl4-challenged and treated animals when compared with prolonged Na-pentobarbitone-induced sleeping time observed in untreated CCl4-intoxicated rats. The methanol extracts of C. filiformis (150 and 300 mg kg−1) were also able to reduce hepatic tissue injury as revealed by a reduction in the serum levels of AST, ALT, ALP, and total bilirubin significantly (p < 0.05) but increased total serum protein significantly (p < 0.05) relative to values in untreated CCl4-induced liver damaged rats. Exposure of rat erythrocytes to varying saline concentrations (0.2–1.0 mg mL−1) of the extract did not produce observable measured increase in osmotic fragility in the red blood cell membranes. The findings suggest that the methanol extract of the aerial parts of C. filiformis could be useful in protecting hepatocytes from toxins especially from alcohol intoxication.

Thermostimulated structural changes in C60-CCl4 and C70-CCl4 crystal solvates

The crystal solvates in C60-CCl4 and C70-CCl4 systems obtained by crystallization from CCl4 solution at room temperature were studied by IR Fourier spectroscopy and X-ray diffraction. The C70 · (CCl4)n crystal solvate was obtained and studied for the first time; its hexagonal crystal lattice has unit cell parameters a = 10.658 ± 0.001 A and c = 10.811 ± 0.002 A. The temperature ranges of its destruction were determined.

The novel Janus kinase inhibitor ruxolitinib confers protection against carbon tetrachloride-induced hepatotoxicity via multiple mechanisms

Therapeutic targeting of the JAK/STAT pathway, the principal signaling mechanism for numerous cytokines, might be an effective approach for limiting inflammation in different organs, including the liver. Therefore, we investigated whether targeting this pathway by the novel JAK inhibitor ruxolitinib could mitigate hepatic damage provoked by carbon tetrachloride (CCl4). Male mice received ruxolitinib treatments (75 and 150mg/kg, oral) 2h prior to intoxication with CCl4 (10ml/kg of 0.3% v/v CCl4 solution in olive oil, intraperitoneal) for 24h. Our results showed that ruxolitinib treatments dose-dependently alleviated CCl4-induced hepatic injury and necroinflammation, as indicated by biochemical markers of injury and histopathology. We unraveled also the mechanisms involved in these hepatoprotective effects. These comprise (i) reducing infiltration of neutrophils and macrophages, as demonstrated by reducing myeloperoxidase activity and F4/80 positive macrophages; (ii) abating apoptosis of hepatocytes, as denoted by decreasing hepatocytes positive for Bax protein; (iii) inhibiting elevation of TNF-α, IL-1β and IL-10; (iv) inhibiting NF-κB activation and translocation to the nucleus, as visualized immunohistochemically; (v) attenuating activation of the IL-23/IL-17 pathway via targeting IL-17, but not IL-23; (vi) antagonizing hepatic oxidative stress by increasing the antioxidant levels (reduced glutathione, glutathione-S-transferase and superoxide dismutase) and decreasing products of lipid peroxidation (malondialdehyde and 4-hydroxynonenal) and total nitrate/nitrite; and (vii) more interestingly, modulating hepatocyte regeneration according to the extent of damage, as quantified by PCNA-immunohistochemistry. In conclusion, our study sheds light on the therapeutic usefulness and the potential underlying mechanisms of the novel JAK inhibitor ruxolitinib in hepatic inflammatory disorders.

Fasting ameliorates metabolism, immunity, and oxidative stress in carbon tetrachloride-intoxicated rats

Fasting has been recently discovered to improve overall health, but its beneficial effects in the presence of hepatic insufficiency have not been proven. The influence of fasting on the metabolism, immunological aspects, and oxidative stress of 40 male carbon tetrachloride (CCl4)-intoxicated Wistar rats was investigated in the present study. The rats were divided into four groups, including a placebo group, CCl4-intoxicated rats, which were injected subcutaneously with 1.0 ml/kg of CCl4 solution, a fasting group, which was fasted 12 h/day for 30 days, and a fourth group, which was injected with CCl4 and fasted. The metabolism, immunity, and oxidative stress improved in CCl4-intoxicated rats fasted for 12 h/day for 30 days, as evidenced in significant increase (p < 0.05) in total protein, globulin, immunoglobulin M (IgM) and IgG levels, and total antioxidant capacity. In contrast, significant decrease (p < 0.05) in blood glucose, total cholesterol, low-density lipoprotein-cholesterol, alanine aminotransferase, C-reactive protein, and malondialdehyde levels were observed. Compared with CCl4-intoxicated rats, significant differences in the albumin, triacylglycerol, high-density lipoprotein-cholesterol, very low-density lipoprotein-cholesterol, cardiovascular risk factor, calcium and magnesium levels were not detected. The results of the present study showed that fasting improved metabolism, immunity, and oxidative stress in CCl4-intoxicated rats. Thus, fasting during Ramadan is safe for patients with hepatic disorders, as the prophet Mohammed (S) said "Keep the fast, keep your health".

Rotational Isomers, Intramolecular Hydrogen Bond, and IR Spectra of o-Vinylphenol

Absorption bands of OH stretching vibrations in IR spectra of o-vinylphenol (o-VP) in the weakly polar solvents CCl4 and n-hexane were studied. Several rotamers of the free OH group were observed for o-VP in n-hexane. The fraction of o-VP rotamers with an O-H…�a intramolecular hydrogen bond (IHB) was less than 20% according to experimental estimates for CCl4 solutions and calculations in the gas phase and cyclohexane. The theoretical effective enthalpy of the o-VP IHB was estimated for rotamer A (-�˚ H = 0.20 kcal/mol). basicity of the �a -bond and the geometric cono guration of the functional groups involved in forming the H-bond. The in? uence of substituents on the IHB strength between the OH group and the double-bond �a -electrons was studied by IR spectroscopy using nine o-VPs with variously substituted vinyl groups as examples (4). It was found that two bands were observed as a rule in the high-frequency region of IR spectra of the o-VPs. These were �� (OH)f in the range 3609-3612 cm -1 and �� (OH)b at 3524-3557 cm -1 . The quantity �˚�� (OH) varied in the range 55-85 cm -1 . The intensity ratio of these bands depended strongly on the position and number of alkyl groups in the vinyl substituent. The compound o-VP itself was studied experimentally (4, 5). The �� (OH)f band in IR spectra of its CCl4 solutions was observed at 3611 cm -1 ; �� (OH)b, 3557 with �˚�� (OH) = 54 cm -1 . IR spectra of the o-phenols were examined only in the range 3700-3500 cm -1 . Any absorption bands in the range 3500-3200 cm -1 were not reported (3-5). The enthalpy of the IHB in o-VP was not determined previously although an estimate of this parameter for its homolog o-isopropenylphenol was -�˚ H = 0.76 ± 0.11 kcal/mol (6). A conformational analysis of structurally ? uxional o-VP was recently completed (7). An analysis of the surface potential energy (SPE) of o-VP that was constructed using the B3LYP/6-311G(d) method showed that o-VP existed in the gas phase and in CCl4 solution as a mixture of rotamers A, B, C, and D that were produced by internal rotation of the OH and vinyl groups and three optical isomers A1, B1, and C1 of these rotamers:

Infrared Spectroscopy of Hydrogen Bonds in Benzoic Acid Derivatives

We have measured the Fourier transform IR spectra of CCl4 solutions of benzoic acid and its biologically active derivatives. We investigated the proton-acceptor properties of the studied molecules theoretically by the molecular electrostatic potential method. The calculations are compared with experimental results. Based on an estimate of the proton-acceptor properties, we give an interpretation of the specific features of the IR spectra of benzoic acid and its derivatives in the region of the O–H and C = O vibrations. The mechanisms for interactions of the molecules are determined by the nature of substituents which are added to the benzene ring in positions para and meta to the carboxyl group. We identify the conditions for appearance of intermolecular hydrogen bonds of O–H · · · O = C, O–H · · · O–H types with formation of cyclic and linear dimers. We show that intramolecular hydrogen bonds of the type O–H · · · O–CH3 prevent the hydroxyl groups from participating in intermolecular interactions.

Dependence of the adsorption of chiral compounds on their enantiomeric composition

The adsorption of two different chiral molecules on platinum surfaces has been explored as a function of enantiomeric composition. In the first case, the saturation monolayers obtained by the adsorption of 1-(naphthyl)ethylamine (NEA) from CCl4 solutions were characterized in situ by reflection–absorption infrared spectroscopy (RAIRS). It was found that racemic mixtures yield different IR spectra than those obtained from enantiopure monolayers, a behavior that was interpreted as the result of the formation of racemate pairs via hydrogen bonding at the amine moiety also responsible for bonding to the surface. NEA adsorption under these conditions is reversible and can be modified by subsequent exposures to solutions of different chiral compositions, but that appears to take place only via changes in the relative fractions of enantiopure versus racemic domains on the surface; no other enantiomeric ratios are apparent in the IR data. The second study focused on the uptake of propylene oxide (PO) on Pt(111) under ultrahigh vacuum (UHV) conditions. In that case, racemic monolayers show densities up to ~20% lower than those obtained with one single enantiomer. This can be explained by kinetic arguments, since data from isothermal molecular-beam experiments indicated that the PO sticking coefficient depends on the chirality of the incoming PO molecules relative to that of the neighboring adsorbed PO species. Monte Carlo simulations could reproduce the experimental data by assuming adsorbate-assisted adsorption and enantiospecific adsorption geometries for molecules impinging on sites adjacent to previously adsorbed surface species.

A Perylene Bisimide Organogelator for Chlorinated Solvents

AbstractDye‐incorporated gel‐like materials are an attractive functional soft matter for optoelectronic applications. Chlorinated solvents are commonly used for the development of organic devices. Herein, we report that a perylene‐bisimide‐functionalized flexible bisurea can work as an efficient organogelator for several chlorinated solvents. The bisurea formed organogels in CHCl3 and 1,1,2,2‐tetrachloroethane (TCE), but it gave viscous solutions in CCl4 at millimolar concentrations. UV/vis and fluorescence spectroscopies, atomic force microscopy, and transmission electron microscopy observation of the diluted solutions revealed that the bisurea forms soluble supramolecular polymers in CCl4 whereas it exists as monomeric or at least oligomeric species in CHCl3 and TCE at micromolar concentrations. These results suggest that differences in the aggregation process could explain the gelation behavior in chlorinated solvents. By using a viscous CCl4 solution, we fabricated top contact organic field effect transistors. The devices showed n‐type semiconductivity with a mobility of 1.5×10−5 cm2 V−1 s−1.

Ameliorative Action of Mn-Salen Derivatives on CCl&amp;lt;sub&amp;gt;4&amp;lt;/sub&amp;gt;-Induced Destructive Effects and Lipofuscin-Like Pigment Formation in Rats’ Liver and Brain: Post-Treatment of Young Rats with EUKs

Lipofuscin-like pigments (LFPs) are highly oxidized cross-linked aggregates of oxidized protein and lipids which are formed under oxidative state conditions by free radicals produced. The present study aimed to evaluate the probable ameliorative effects of some of the Mn-salens namely EUKs 8, 134, 15, 115, 122 and 132 (compounds 1-6) and vitamin C against carbon tetrachloride (CCl4)-induced acute damage on rats’ livers and brains. Exposure to CCl4 is believed to induce oxidative stress and cause tissue damage due to the formation of trichloromethyl (·CCl3) and peroxy trichloromethyl (·OOCCl3) radicals. In this study, 54 rats were randomly divided into 9 groups of six each: normal group received only vehicle (olive oil; 2 ml/kg b.w.) for 6 consecutive days; CCl4- intoxicated group received the vehicle and CCl4 (50% solution of CCl4 in olive oil, 2 ml/kg b.w.) on the first and second days and the vehicle on the third to sixth days; test rats received Mn-salens or vitamin C (20 mg/kg b.w.) and CCl4 (2 ml/kg b.w.) on the first and second days and Mn-salens or vitamin C (20 mg/kg b.w.) on the third to sixth days. Mn-salens administration ameliorated the effects of CCl4 by decreasing the levels of ROS, lipid and protein oxidations and LFPs formation on liver and brain as well as cholesterol and triglycerides, aminotransferases and alkaline phosphatase contents in sera of rats whereas increased the activities of catalase, superoxide dismutase,glutathione reductase, glutathione peroxidase and reduced glutathione in liver and brain tissues. Histopathological studies confirmed the toxic effects of CCl4 and ameliorative action of Mn-salens on tissues. These results suggest that the evaluated EUKs were able to attenuate LFPs accumulation and morphological changes caused by CCl4 in rats and thus, confirming the ameliorative role of Mn-salens against CCl4-induced oxidative damage and age-related diseases.

Anti-fibrosis effects of Huisheng oral solution in CCl4-induced hepatic fibrosis in rat.

Aim:Some gradient of Huisheng oral solution (HOS) has been reported to have anti-fibrosis activity. This study was designed to investigate whether HOS could inhibit liver fibrosis and to elucidate its molecular mechanism of action.Materials and Methods:Hepatic fibrosis model in rat was induced by subcutaneous injection of CCl4. Rats in the treatment group were administrated with HOS intragastrically. Hematoxylin and eosin (H and E) staining and Masson's trichrome staining were used to examine the changes in liver pathology. Levels of ALT, AST, LDH, hyaluronic acid (HA) and laminin (LN) in serum and hydroxyproline (Hyp) in liver were detected by biochemical examination and radioimmunoassay, respectively. The expression and distribution of Smad3, TGF-β1, α-SMA and TIMP-1 were observed and the active TGF-β1 was tested.Results:Our data demonstrated that HOS alleviated CCl4-induced collagen deposition in liver tissue, improved liver condition and liver function in rats. HOS also significantly reduced the expression and distribution of Smad3, TGF-β1, α-SMA and TIMP-1 as well as decreased active TGF-β1.Conclusions:This study revealed that HOS attenuates the development of liver fibrosis through suppressing the TGF-β1 pathway. It provides us a new approach to treatment of liver fibrosis.

Experimental Study on the Effect of Different Pretreatment Methods on Rubber Concrete Work Performance and Compressive Strength

In untreated, clean water, NaOH solution to clean, CCl4solution to clean the rubber particles of different pretreatments as the main influencing factors, by 51 rubber concrete block pilot study reached different pretreatment methods on different rubber particle size and dosage of rubber concrete workability and compressive strength were investigated. The results show that: water, NaOH solution, CCl4pretreatment solution, such as a rubber concrete workability and compressive strength were improved, and the rubber particles larger pretreatment on rubber compressive strength of concrete work to improve performance and more obvious.

Hydrogen-Bond Acidity of OH Groups in Various Molecular Environments (Phenols, Alcohols, Steroid Derivatives, and Amino Acids Structures): Experimental Measurements and Density Functional Theory Calculations

The hydrogen-bond (H-bond) donating strengths of a series of 36 hydroxylic H-bond donors (HBDs) with N-methylpyrrolidinone have been measured in CCl4 solution by FTIR spectrometry. These data allow the definition of a H-bond acidity scale named pKAHY covering almost three pK units, corresponding to 16 kJ mol(-1). These results are supplemented by equilibrium constants determined in CH2Cl2 for one-third of the data set to study compounds showing a poor solubility in CCl4. A systematic comparison of these experimental results with theoretical data computed in the gas phase using DFT (density functional theory) calculations has also been carried out. Quantum electrostatic parameters appear to accurately describe the H-bond acidity of the hydroxyl group, whereas partial atomic charges according to the Merz-Singh-Kollman and CHelpG schemes are not suitable for this purpose. A substantial decrease of the H-bond acidity of the OH group is pointed out when the hydroxyl moiety is involved in intramolecular H-bond interactions. In such situations, the interactions are further characterized through AIM and NBO analyses, which respectively allow localizing the corresponding bond critical point and the quantification of a significant charge transfer from the available lone pair to the σ*OH antibonding orbital. Eventually, the H-bond ability of the hydroxyl groups of steroid derivatives and of lateral chains of amino acids are evaluated on the basis of experimental and/or theoretical data.

Synthesis, Properties, and Structures of Chromium(VI) and Chromium(V) Complexes with Heterocyclic Nitrogen Ligands

AbstractThe reaction of CrO2Cl2 with 2, 2′‐bipyridyl or 1, 10‐phenanthroline (diimine) in CCl4 or anhydrous CH3CO2H solution, produces orange‐brown diamagnetic [CrO2Cl2(diimine)]. The X‐ray structure of [CrO2Cl2(2, 2′‐bipy)] shows a six‐coordinate central chromium(VI) atom with cis‐dioxo groups trans to the diimine. In contrast, the diimines react with CrO3 in CH3CO2H / conc. aqueous HCl to form bright red paramagnetic CrV complexes, [CrOCl3(diimine)]. The X‐ray structure of [CrOCl3(2, 2′‐bipy)] shows a six‐coordinate central chromium atom with mer‐chlorines and the diimine trans to O/Cl. The addition of [2, 2‐bipyH2]Cl2 to a solution of CrO3 in CH3CO2H saturated with HCl gas, produces the CrV anion [2, 2′‐bipyH2][CrOCl4]Cl, which loses HCl on heating in vacuo to form [CrOCl3(2, 2′‐bipy)]. IR, UV/Vis, and 1H NMR spectra (CrVI only) are reported for the new complexes. Attempts to extend these routes to oxygen donor ligands, including ethers and phosphine oxides, were unsuccessful. The diimine complexes are the first structurally autheticated adducts of chromium(VI) and (V) oxide‐chlorides with neutral ligands.

Mass Transport across the Porous Oxide Shells of Core–Shell and Yolk–Shell Nanostructures in Liquid Phase

The mass transport of a number of molecules dissolved in a liquid solvent through the shells of Pt@SiO2 core–shell and Pt@Void@TiO2 yolk–shell nanostructures was characterized in situ by infrared absorption spectroscopy. Our samples, which were used here to represent the core–shell and yolk–shell nanostructures that have become so popular in recent times, were determined to exhibit a dual distribution of pore sizes, with a majority of micropores with diameters of less than 1 nm and a second group of mesopores with windows approximately 4 nm in diameter. The uptake of carbon monoxide from CCl4 or ethanol solutions onto the surface of the metal of these nanostructures was characterized first. It was determined that adsorption is possible with both samples, and that saturation on the metal surface precedes saturation on the oxide shell. This latter observation suggests that the access of the CO molecules (and all other molecules studied here) to the inside of the yolk–shell structures may be controlled by the mass transport of the liquid through the porous network of the shells, not by the diffusion of the dissolved gas alone. Additional studies were carried out with a family of cinchona alkaloid derivatives and with a porphyrin, covering a range of molecular sizes of approximately 5–20 Å in diameter. Adsorption of all those molecules on the Pt surfaces of both nanoarchitectures was deemed possible by a combination of tests, including the detection of changes in peak frequencies or peak intensities and measurements of the reversibility of the adsorption. Our results indicate that although the sol–gel synthetic method used to prepare the shells usually produces solids with microporous structures, a secondary mesoporous network that also develops in these nanostructures can provide a direct path to the metal nanocores, affording the mass transport of large molecules in liquid solutions in and out of the inside volumes. We also show that the attoliter-size volumes encased by typical shells in yolk–shell nanostructures are sufficient to afford adsorption displacement processes.

Intermolecular C–H⋯O interactions in cyclopentanone: An inelastic neutron scattering study

The inelastic neutron scattering (INS) spectra of cyclopentanone were obtained for pure and 50% CCl4 solution forms. Spectra are compared with infrared and Raman data, and with DFT calculated eigenvectors. This exercise aims to find spectroscopic evidence in the neutron spectra for the presence of C–H⋯O hydrogen bonds. These are weak interactions with an energy of ca. −6kJmol−1 as predicted by DFT. The neutron spectra show narrow and sharp bands which allows for an assignment of the vibrational modes. The simulated neutron spectrum of C–H⋯O bonded cyclopentanone dimers matches the experimental spectrum of the pure compound, whereas the monomer simulation monomer matches the experimental spectrum of the diluted solution, meaning that such interaction can be probed by INS. Assignment of the 95cm−1 band to the νH⋯O anti-translational mode, being supported by DFT results and in agreement with previous literature data, is considered and discussed.

Is There a Simple Way to Reliable Simulations of Infrared Spectra of Organic Compounds?

To assess the ability of the quantum-chemical computations to reproduce the experimental relative intensities in the infrared (IR) spectra of both the gas- and condensed-phase systems, the hybrid DFT functional B3LYP has been applied to simulation of IR spectra for species containing from three to twelve first- or second-row atoms, both in the gas phase and in CCl4 solutions. The results demonstrate that B3LYP, combined with the highly compact double-ζ basis set 6-31+G* and "scaled quantum mechanics" techniques, offers excellent quantitative performance in the calculations of relative IR intensities and frequencies (ν ≤ 2200 cm(-1)) for the bands of vibrations of medium-size isolated molecules, whereas it produces unsatisfactory results for the solutions of the same species. Neither larger basis sets nor implicit treatment of the media effects improve the agreement of the simulated spectra with the condensed-phase experiment. At the same time, some preliminary results suggest that explicit modeling of media effects could offer better quality of the IR spectral simulations for the condensed-phase systems.

Morphological features of C60 and C70 microcrystals obtained from C6H5CH3 and CCl4 solutions

Fullerene C60 and C70 microcrystals of different morphologies obtained by crystallization from toluene and carbon tetrachloride solutions were investigated using scanning electron microscopy and diffuse reflectance IR Fourier transform spectroscopy. The obtained results are discussed with regard to the features of fullerenes clusterization processes in solutions.

Glycyrrhizic acid attenuates CCl4-induced hepatocyte apoptosis in ratsviaa p53-mediated pathway

To investigate the effect of glycyrrhizic acid (GA) on carbon tetrachloride (CCl4)-induced hepatocyte apoptosis in rats via a p53-dependent mitochondrial pathway. Forty-five male Sprague-Dawley rats were randomly and equally divided into three groups, the control group, the CCl4 group, and the GA treatment group. To induce liver fibrosis in this model, rats were given a subcutaneous injection of a 40% solution of CCl4 in olive oil at a dose of 0.3 mL/100 g body weight biweekly for 8 wk, while controls received the same isovolumetric dose of olive oil by hypodermic injection, with an initial double-dose injection. In the GA group, rats were also treated with a 40% solution of CCl4 plus 0.2% GA solution in double distilled water by the intraperitoneal injection of 3 mL per rat three times a week from the first week following previously published methods, with modifications. Controls were given the same isovolumetric dose of double distilled water. Liver function parameters, such as alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were determined. Pathologic changes in the liver were detected by hematoxylin and eosin staining. Collagen fibers were evaluated by Sirius red staining. Hepatocyte apoptosis was investigated using the terminal deoxynucleotidyl transferase-mediated deoxyuridine 5-triphosphate nick end labeling (TUNEL) assay and the cleaved caspase-3 immunohistochemistry assay. The expression levels of p53 and apoptosis-related proteins were evaluated by immunohistochemistry or Western blotting analysis. After 8 wk of treatment, GA significantly reduced serum activity of ALT (from 526.7 ± 57.2 to 342 ± 44.8, P < 0.05) and AST (from 640 ± 33.7 to 462.8 ± 30.6, P < 0.05), attenuated the changes in liver histopathology and reduced the staging score (from 3.53 ± 0.74 to 3.00 ± 0.76, P < 0.05) in CCl4-treated rats. GA markedly reduced the positive area of Sirius red and the ratio of the hepatic fibrotic region (from 7.87% ± 0.66% to 3.68% ± 0.32%, P < 0.05) compared with the CCl4 group. GA also decreased the expression level of cleaved caspase-3 compared to the CCl4 group. TUNEL assay indicated that GA significantly diminished the number of TUNEL-positive cells compared with the CCl4 group (P < 0.05). GA treatment clearly decreased the level of p53 (P < 0.05) detected by immunohistochemistry and Western blotting analysis. Compared with the CCl4 group, we also found that GA reduced the Bax/Bcl-2 ratio (P < 0.05), the expression of cleaved caspase-3 (P < 0.05), cleaved caspase-9 (P < 0.05), and inhibited cytochrome C and second mitochondria-derived activator of caspases (Smac) release from mitochondria to cytoplasm, i.e., GA reduced the expression level of Smac, which inhibited c-IAP1 activity (P < 0.05), ultimately inhibiting the activity of caspase-3, according to Western blotting analysis. As a result, GA suppressed activation of the caspase cascades and prevented hepatocyte apoptosis. GA can inhibit CCl4-induced hepatocyte apoptosis via a p53-dependent mitochondrial pathway to retard the progress of liver fibrosis in rats.

Dynamic Molecular Behavior and Cluster Structure of Octanoic Acid in Its Liquid and CCl&amp;lt;sub&amp;gt;4&amp;lt;/sub&amp;gt; Solution

Fatty acids such as oleic and stearic acids having a long hydrocarbon chain are known to exist as dimers in their melt and even in a non-polar solvent. In their melt the dimers arrange longitudinally and alternately to form clusters which resemble a smectic liquid crystal. The clusters determine the liquid properties of the fatty acids such as density, viscosity and fluidity. Then, do the dimers of fatty acid having a moderate-length hydrocarbon chain construct such the clusters? In the present study the dynamic molecular behavior and assembly structure of octanoic acid in its melt and also in CCl4 solution have been investigated by the X-ray diffraction, near infrared spectroscopy, 1H-NMR chemical shift, self-diffusion coefficient and 13C-NMR spin-lattice relaxation time measurements. From these results it has been revealed that the clusters of octanoic acid exist in its melt and also in CCl4 and that the clusters in the melt disintegrate with an increase in temperature. The dissociation profile of dimers of octanoic acid into monomers in CCl4 also has been clarified.

Fuzheng Huayu inhibits carbon tetrachloride-induced liver fibrosis in mice through activating hepatic NK cells

Aim of the studyFuzheng Huayu (FZHY) is a Chinese compound herbal preparation which consists of six Chinese herbs. This study examines the preventative effects of FZHY on liver fibrosis induced by carbon tetrachloride (CCl4) and explores its possible mechanisms of action. Materials and methodsLiver fibrosis was induced in male C57BL/6N mice by injecting a 10% CCl4 solution intraperitoneal twice a week for six weeks. After 6 weeks of treatment, serum ALT and AST assay, liver tissue histological examination and immunostaining were carried out to examine the liver function and fibrosis degree. The expression levels of alpha-smooth muscle actin (SMA) were measured by quantitative real-time PCR and western blot. Hepatic natural killer (NK) cells were isolated from liver and evaluated by FACS. ResultsUpon pathological examination, the FZHY-treated mice showed significantly reduced liver damage. The expression of α-SMA increased markedly upon treatment with CCl4 and the increase was reversed by FZHY treatment. FZHY treatment also enhanced the activation of hepatic NK cells and the production of interferon-gamma (IFN-γ). The protective effects of FZHY were reversed in the mice that were depleted of NK cells by anti-ASGM-1 Ab treatment. ConclusionsFZHY can efficiently inhibit CCl4-induced liver fibrosis. Furthermore, the depletion of NK cells attenuates the protective effects of FZHY. We conclude that FZHY could be an effective drug for liver fibrosis, and its mechanism of action involves the activation of hepatic NK cells.