Chemical and Biological Evaluation of Bakeries Produced from Golden Berries

This investigation was performed with the purpose of researching the influence of pizza containing dried golden berry fruits (DGBF) at different doses against carbon tetrachloride - induced hepatotoxicity in rats. The study shows phenols content of golden berry. 25 male rats were used in the biological investigation. Rats were divided into five groups (5 rats in group) the investigation was 12 weeks. The first group (negative group) was given a basal diet and the second group (G2, G3, G4, and G5) was injected intramuscularly with carbon tetrachloride 2 ml/kg BW (50% v/v in liquid paraffin) weekly to induce hepatotoxicity. After the injury, group G3, G4 and G5 fed on 50% basal diet supplemented with 50%pizza containing 5, 10 and 15% DGBF. Findings indicate that DGBF had a high antioxidant activity, total phenol, flavonoids, ascorbic acid and carotenoids content. Rats fed 50% pizza containing (5,10 and 15%) DGBF had a lower serum total cholesterol, LDL cholesterol, triglyceride, urea, uric acid, creatinine, GOT, GPT, MDA and SOD compared to rats fed simply the basal diet (positive control). The DGBF was added to the pizza with different proportions, and its sensory properties were evaluated, and all proportions were proper to the panelists, compared to the control. The findings of this work suggest that golden berries could be used to treat and prevent hepatotoxicity patients.

Acutely applied melatonin prevents CCl4-induced testicular lesions in rats: the involvement of the oxidative capacity and arginine metabolism

Abstract Carbon tetrachloride (CCl4) represents an organic chemical that causes reactive oxygen species derived organ disturbances including male infertility. Melatonin (MLT) is a neurohormone with strong antioxidant capacity, involved in numerous physiological processes. In this study we evaluated the capability of MLT, administered in a single dose of 50 mg/kg, to preserve the testicular tissue function after an acute administration of CCl4 to rats. The disturbance in testicular tissue and the effects of MLT after CCl4 exposure were estimated using biochemical parameters that enabled us to determine the tissue (anti)oxidant status and the intensity of arginine/nitric oxide metabolism. Also, the serum levels of testosterone and the histopathological analysis of tissue gave us a better insight into the occurring changes. A significant diminution in tissue antioxidant defences, arginase activity and serum testosterone levels, followed by the increased production of nitric oxide and extensive lipid and protein oxidative damage, was observed in the CCl4-treated group. The application of MLT after the CCl4 caused changes, clearly visible at both biochemical and histological level, which could be interpreted mainly as a consequence of general antioxidant system stimulation and a radical scavenger. On the other hand, the application of MLT exerted a limited action on the nitric oxide signalling pathway.

Effect of Acute Liver Injury on Pharmacokinetics and Tissue Distribution of Baicalin Magnesium in Rats / 中国实验方剂学杂志

ObjectiveTo establish a high performance liquid chromatography （HPLC） for simultaneous determination of baicalin magnesium and baicalein in rat plasma and tissues， and to investigate the effect of acute liver injury on pharmacokinetics and tissue distribution of baicalin magnesium in rats. MethodAcute liver injury rat model was induced by carbon tetrachloride （CCl4）. Normal rats and acute liver injury model rats were given an equal dose （287.31 mg·kg-1） of baicalin magnesium aqueous solution by intragastric administration， the orbital blood was collected at different time points， and HPLC was used to simultaneously determine the concentrations of baicalin magnesium and baicalein in rat plasma at each time point， the concentration-time curves were drawn， the pharmacokinetic parameters were calculated with DAS 3.0， and SPSS 23.0 was used for statistical analysis. After oral administration of baicalin magnesium aqueous solution， HPLC was used to simultaneously determine the contents of baicalin magnesium and baicalein in rat liver， lung， kidney， stomach， brain and small intestine at different time points， the mobile phase was 0.1% phosphoric acid aqueous solution-methanol， and the detection wavelength was 278 nm. ResultIn the acute liver injury model group， the peak concentration （Cmax） of baicalin magnesium was 0.58 times that of the normal group， the area under concentration-time curve （AUC0-t） was 0.5 times that of the normal group （P<0.05）， the apparent volume of distribution （Vd） was 2.3 times that of the normal group （P<0.05）， and baicalein is almost undetectable in plasma. The content of baicalin magnesium in liver， stomach and brain of the acute liver injury model group was higher than that of the normal group at each time point， while the content of baicalin magnesium in the samples of lung at 8 h， kidney at 8 h and 12 h， and small intestine at 0.333 h was lower than that of the normal group. The content of baicalein in lung， stomach and small intestine of the model group was higher than that of the normal group at each time point， while the content of baicalein in the tissue samples of liver at 6， 8 h and kidney at 0.333， 4， 6 h was lower than that in the normal group， and baicalein could hardly be detected in the brain. ConclusionAfter intragastric administration of the same dose of baicalin magnesium aqueous solution， acute liver injury induced by CCl4 can affect the pharmacokinetics and tissue distribution characteristics of baicalin magnesium in rats， and there is biotransformation of baicalin magnesium and baicalein in liver， lung， kidney， stomach and small intestine.

Mechanism of Zhizi Prescription in Protection of CCl4-induced Acute and Subacute Liver Injury in Mice / 中国实验方剂学杂志

ObjectiveTo investigate the protective effect of Zhizi prescription （ZZP） on carbon tetrachloride （CCl4）-induced acute and subacute liver injury and its mechanism. MethodAcute and subacute liver injury animal models were induced. C57 mice were randomly divided into a normal group， model group， obeccholic acid group， ZZP high-dose （0.5 g·kg-1） group， and ZZP low-dose （0.25 g·kg-1） group. According to the experiment design， the serum and liver tissue of mice were collected after the last administration. Hematoxylin-eosin （HE） and Sirius staining was used to observe the liver pathological changes. Serum alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， total bilirubin （TBIL）， liver homogenate hydroxyproline （Hyp）， malondialdehyde （MDA）， and superoxide dismutase （SOD） levels were determined by kit. The levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in the liver tissue were determined by enzyme-linked immunosorbent assay （ELISA）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expressions of collagen 1A1 （Col1a1）， collagen 3A1 （Col3a1）， fibronectin （FN）， transforming growth factor β receptor Ⅱ （Tgfbr2） and α-smooth muscle actin （α-SMA） in the liver tissue. ResultIn terms of the acute liver injury， as compared with the normal group， the levels of ALT， AST， TBIL and MDA in the model group were significantly increased （P<0.01）， while the activity of liver SOD was significantly decreased （P<0.01）. Compared with model group， the ZZP high-dose and low-dose groups both significantly reduced the degree of liver cell injury， and protected the acute liver injury induced by CCl4. The ZZP high-dose group had a better effect than the ZZP low-dose group. In terms of the subacute liver injury， the levels of ALT， AST， MDA，TNF-α and IL-6 in the model group were significantly increased （P<0.01）， while the activity of liver SOD was significantly decreased （P<0.01）. As compared with the model group， liver Hyp content in the ZZP high-dose and low-dose groups was significantly decreased （P<0.01）， and the collagen deposition in liver of both groups was significantly reduced. The ZZP high-dose group also significantly down-regulated the mRNA expressions of α-SMA， Col1a1， Col3a1， FN， and Tgfbr2 in the liver of mice （P<0.05， P<0.01）. ConclusionZZP effectively protects the acute and subacute liver injury induced by CCl4， and the protective effect is proportional to its concentration. The mechanism may be related to the increase of the activity of antioxidant enzymes in the liver tissue， the decrease of the level of lipid peroxidation， and the inhibition of inflammatory response， thus reducing collagen deposition and improving early liver fibrosis.

Mechanism of Zhizi Prescription in Protection of CCl4-induced Acute and Subacute Liver Injury in Mice / 中国实验方剂学杂志

ObjectiveTo investigate the protective effect of Zhizi prescription （ZZP） on carbon tetrachloride （CCl4）-induced acute and subacute liver injury and its mechanism. MethodAcute and subacute liver injury animal models were induced. C57 mice were randomly divided into a normal group， model group， obeccholic acid group， ZZP high-dose （0.5 g·kg-1） group， and ZZP low-dose （0.25 g·kg-1） group. According to the experiment design， the serum and liver tissue of mice were collected after the last administration. Hematoxylin-eosin （HE） and Sirius staining was used to observe the liver pathological changes. Serum alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， total bilirubin （TBIL）， liver homogenate hydroxyproline （Hyp）， malondialdehyde （MDA）， and superoxide dismutase （SOD） levels were determined by kit. The levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in the liver tissue were determined by enzyme-linked immunosorbent assay （ELISA）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expressions of collagen 1A1 （Col1a1）， collagen 3A1 （Col3a1）， fibronectin （FN）， transforming growth factor β receptor Ⅱ （Tgfbr2） and α-smooth muscle actin （α-SMA） in the liver tissue. ResultIn terms of the acute liver injury， as compared with the normal group， the levels of ALT， AST， TBIL and MDA in the model group were significantly increased （P<0.01）， while the activity of liver SOD was significantly decreased （P<0.01）. Compared with model group， the ZZP high-dose and low-dose groups both significantly reduced the degree of liver cell injury， and protected the acute liver injury induced by CCl4. The ZZP high-dose group had a better effect than the ZZP low-dose group. In terms of the subacute liver injury， the levels of ALT， AST， MDA，TNF-α and IL-6 in the model group were significantly increased （P<0.01）， while the activity of liver SOD was significantly decreased （P<0.01）. As compared with the model group， liver Hyp content in the ZZP high-dose and low-dose groups was significantly decreased （P<0.01）， and the collagen deposition in liver of both groups was significantly reduced. The ZZP high-dose group also significantly down-regulated the mRNA expressions of α-SMA， Col1a1， Col3a1， FN， and Tgfbr2 in the liver of mice （P<0.05， P<0.01）. ConclusionZZP effectively protects the acute and subacute liver injury induced by CCl4， and the protective effect is proportional to its concentration. The mechanism may be related to the increase of the activity of antioxidant enzymes in the liver tissue， the decrease of the level of lipid peroxidation， and the inhibition of inflammatory response， thus reducing collagen deposition and improving early liver fibrosis.

Transformation mechanism of carbon tetrachloride and the associated micro-ecology in landfill cover, a typical functional layer zone / 生物工程学报

Landfill is one of the important sources of carbon tetrachloride (CT) pollution, and it is important to understand the degradation mechanism of CT in landfill cover for better control. In this study, a simulated landfill cover system was set up, and the biotransformation mechanism of CT and the associated micro-ecology were investigated. The results showed that three stable functional zones along the depth, i.e., aerobic zone (0-15 cm), anoxic zone (15-45 cm) and anaerobic zone (> 45 cm), were generated because of long-term biological oxidation in landfill cover. There were significant differences in redox condition and microbial community structure in each zone, which provided microbial resources and favorable conditions for CT degradation. The results of biodegradation indicated that dechlorination of CT produced chloroform (CF), dichloromethane (DCM) and Cl- in anaerobic and anoxic zones. The highest concentration of dechlorination products occurred at 30 cm, which were degraded rapidly in aerobic zone. In addition, CT degradation rate was 13.2-103.6 μg/(m2·d), which decreased with the increase of landfill gas flux. The analysis of diversity sequencing revealed that Mesorhizobium, Thiobacillus and Intrasporangium were potential CT-degraders in aerobic, anaerobic and anoxic zone, respectively. Moreover, six species of dechlorination bacteria and eighteen species of methanotrophs were also responsible for anaerobic transformation of CT and aerobic degradation of CF and DCM, respectively. Interestingly, anaerobic dechlorination and aerobic transformation occurred simultaneously in the anoxic zone in landfill cover. Furthermore, analysis of degradation mechanism suggested that generation of stable anaerobic-anoxic-aerobic zone by regulation was very important for the harmless removal of full halogenated hydrocarbon in vadose zone, and the increase of anoxic zone scale enhanced their removal. These results provide theoretical guidance for the removal of chlorinated pollutants in landfills.

In vivo protective effect of cinnamon aqueous extract in carbon tetrachloride-treated male albino rats

The liver as an organ is important for the metabolism of drugs and toxins. However, it is not immune from environmental insults. Exposure of liver cells to carbon tetrachloride (CCl4) results in the generation of tricholoromethyl radicals, which induce liver toxicity. This study aims at investigating the ameliorative effect of the cinnamon aqueous extract (CAE) against CCl4-induced hepatotoxicity in male albino rats. Hepatotoxicity was induced in rats through the intraperitoneal administration of 0.5 mL kg-1body weight of CCl4. The analyses of the results obtained showed significant reduction in the levels of serum biochemical markers for 400 and 600 mg kg-1bw of CAE protected rats as compared with CCl4group. In addition, CAE administration reversed liver tissue damaged via increased antioxidants markers. Histopathological examination of CAE treatment on rats showed improved changes to the liver damage caused by CCl4 with no evidence of steatosis and inflammation. This result hence suggests that CAE has marked hepatoprotective and healing activities against CCl4-induced liver damage and could serve as a suitable candidate in drug discovery for the treatment of liver toxicity.

Ameliorative effect of psoralen on acute liver injury induced by carbon tetrachloride in mice / 中国药科大学学报

@#To investigate the ameliorative effect of psoralen (PSO) on carbon tetrachloride (CCl4)-induced acute liver injury in mice and its potential mechanism, female C57BL/6J mice aged 6-8 weeks were continuously administrated with psoralen or positive control drug diallyl sulfide (DAS) intragastrically for 4 days.On day 4, except that the control group were treated with vehicle control, other groups were all given carbon tetrachloride intraperitoneally to establish a carbon tetrachloride acute liver injury model.Serum biochemical indicators alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were detected; liver pathological changes were observed by HE staining; cytochrome P450 2E1 (CYP2E1) protein levels were detected by Western blot; the protein level of CYP2E1 were detected by immunohistochemistry (IHC) staining; and the gene levels of CYP2E1, tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were detected by RT-PCR.Compared with the model group, psoralen could improve the inflammatory cell infiltration and hepatocyte necrosis caused by carbon tetrachloride, significantly reducing the serum ALT and AST levels, down-regulating the inflammatory factors TNF-α and IL-6 levels, and inhibiting CYP2E1 protein expression.The results show that psoralen can ameliorate the acute liver injury induced by carbon tetrachloride in mice, with the possible mechanism inhibiting the protein expression of CYP2E1.

LIX1-like protein promotes liver cancer progression

Limb and CNS expressed 1 like (LIX1L) is over-expressed in several types of tumors. However, the function of LIX1L in glucose metabolism and hepatocellular carcinoma (HCC) progression remains elusive. Here we report that LIX1L is over-expressed in human HCC tissues, which predicts unfavorable prognosis. LIX1L deficiency

Protective Effect of Gentiopicroside Against Liver Injury in Mice via TLR4/MyD88/NF-κB Signaling Pathway / 中国实验方剂学杂志

Objective:To explore the mechanism of gentiopicroside （GPS） in preventing acute liver injury induced by carbon tetrachloride （CCl₄） in mice and its effect on the Toll-like receptor 4 （TLR4）/myeloid differentiation factor 88 （MyD88）/nuclear factor-<italic>κ</italic>B （NF-<italic>κ</italic>B） signaling pathway. Method:Sixty mice were randomly divided into a normal control group， a model group， a silymarin group （150 mg·kg^-1）， and high- （200 mg·kg^-1）， medium- （100 mg·kg^-1）， and low-dose （50 mg·kg^-1） GPS groups， with 10 in each group. The mice in the groups with drug intervention were administered correspondingly by gavage at 10 mL·kg^-1， and those in the normal control group and the model group receive an equal volume of distilled water， once per day. Ten days after administration， mice in the normal control group were subjected to the intraperitoneal injection of peanut oil （10 mL·kg^-1） and those in other groups were injected with peanut oil （10 mL·kg^-1） containing 0.12% CCl₄for the induction of acute liver injury model. After fasting for 16 hours， blood was collected from eyeballs and liver tissues were collected. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of liver tissues. The content or activities of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， total bilirubin （TBIL）， alkaline phosphatase （ALP）， total superoxide dismutase （T-SOD）， and <italic>γ</italic>-glutamyl transpeptidase （<italic>γ</italic>-GT） in the serum， malondialdehyde （MDA） and glutathione peroxidase （GSH-Px） in liver tissues were determined by biochemistry techniques. The levels of tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>）， interleukin-1<italic>β</italic> （IL-1<italic>β</italic>）， and interleukin-6 （IL-6） in liver tissues were measured by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the protein expression of TLR4， MyD88， and NF-<italic>κ</italic>B in liver tissues. The expression of phosphorylated NF-<italic>κ</italic>B （p-NF-<italic>κ</italic>B） was detected by immunohistochemistry. Result:Compared with the normal control group， the model group showed increased levels of ALT， AST， ALP， TBIL， <italic>γ</italic>-GT， and MDA （<italic>P</italic><0.01）， and blunted activities of T-SOD and GSH-Px （<italic>P</italic><0.01）. Compared with the model group， the high- and medium-dose GPS groups exhibited declining levels of ALT， AST， ALP， TBIL， <italic>γ</italic>-GT， and MDA （<italic>P</italic><0.05， <italic>P</italic><0.01） and potentiated T-SOD and GSH-Px activities （<italic>P</italic><0.05， <italic>P</italic><0.01）. Compared with the normal control group， the model group displayed elevated levels of TNF-<italic>α</italic>， IL-1<italic>β</italic>， and IL-6 in liver tissues （<italic>P</italic><0.01） and increased protein expression of TLR4， MyD88， and p-NF-<italic>κ</italic>B （<italic>P</italic><0.01）. Compared with the model group， the high- and medium-dose GPS groups showed decreased TNF-<italic>α</italic>， IL-1<italic>β</italic>， and IL-6 content in liver tissues （<italic>P</italic><0.05， <italic>P</italic><0.01） and dwindled TLR4， MyD88， and p-NF-<italic>κ</italic>B protein expression （<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:GPS possesses a protective effect on mice with acute liver injury induced by CCl₄， and its mechanism of action may be related to the regulation of TLR4/MyD88/NF-<italic>κ</italic>B signaling pathway and inhibition of oxidative stress.

Hepatoprotective Activity Of Ethanolic Stem Bark Extract Of Knema Attenuata (HOO K. F. And Thomson) Warb

Objective: To investigate the hepatoprotective activity of ethanolic stem bark extract (ESBE) of Knema attenuata against carbon tetrachloride (CCl4) induced hepatotoxicity in Wistar rats using both in vivo and in vitro models. Methods: Animals were treated orally with ESBE (250 mg kg-1 and 500 mg kg-1) once daily for 6 d and CCl4 on the 4th d. On the 7th d, animals were sacrificed and the blood samples were collected to measure the serum levels of biochemical parameters, whereas the liver homogenates were utilized for estimating the antioxidant defense. The hepatoprotective efficacy of the extract was further ensured in vitro using human liver hepatocellular carcinoma (HepG2) cell line against CCl4 induced toxicity. The cell line viability was determined using 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Results: ESBE effectively reduced (p<0.001) the elevated serum levels of Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), and Alkaline phosphatase (ALP) when compared to the toxicant control group. ESBE 500 mg kg-1significantly raised the antioxidant defense (p<0.0001) by reducing the malondialdehyde (MDA) level and enhancing hepatic reduced glutathione (GSH) level in comparison to the CCl4 control group. The in vitro effect was investigated using CCl4 exposed HepG2 cells. Pretreatment with ESBE showed a dose-dependent increase in percentage cell viability ranged between 44 to 57% at 12.5-100 μg ml-1concentrations (p<0.001, when compared to the control cells).  Conclusion: Present study confirms the hepatoprotective activity of the stem bark extract of K. attenuata against CCl4‑induced liver damage.

Therapeutic Effect Of Silver Nanoparticles Against Diethyl Nitrosamin And Carbon Tettrachloride-Induced Hepatocellular Carcinoma In Rats

Objective: Hepatic cancer is known as primary liver cancer and hepatocellular carcinoma (HCC). Newly silver nanoparticles gained importance due to its advantages and multiple potential such as molecular imaging agent, antimicrobial, wound healing, anti-inflammatory and anticancer activity. The current study deals to assess therapeutic property silver nanoparticles (AgNPs) against diethylnitrosamine (DENA), and carbon tetrachloride (CCL4) induced hepatic cancer. Methods: Thirty male albino rats (200-250g) were distributed into four groups and hepatic cancer was induced with a single intraperitoneal dose of 200 mg/kg body weight of DENA. Two weeks later, animals received subcutaneous injections of CCl4 once a week in a dose of 3 ml/kg body weight for 6weeks. Serum biomarkers, antioxidants enzymes, inflammatory markers were evaluated to find the anti-proliferative potential of silver nanoparticles. Histological evaluation and microscopic reports were also done to document the results of the current work. Results: AgNPs significantly recover the serum marker enzymes of hepatic parameter AST, ALT, ALP, and total bilirubin and also decreased the levels of NO, IL-6 and TNF-α. Histopathological features also exhibited recovery of a hepatic architecture in cancer-induced rats. Moreover, the immunohistochemical investigation demonstrated that the levels of PCNA, and Caspase-3, which are hepatocarcinogenic markers, were significantly improved by AgNPs. Conclusion: These results concluded that AgNPs showed promising curing effects on hepatocellular ailments.

Bidens pilosa L. (Asteraceae) cultivated in Brazil on acute liver disease in dogs / [Bidens pilosa L. (Asteraceae) cultivada no Brasil na doença hepática aguda em cães]

Bidens pilosa L. is a medicinal plant popularly used for treatment of liver diseases. In this study, the dry extract of aerial parts of Bidens pilosa and Silymarin, a phytocomplex obtained from the Silybum marianum fruits and marketed as hepatoprotective, were tested in dogs experimentally acutely intoxicated with carbon tetrachloride. The liver activity was evaluated by hematological and biochemical profiles, and histological and ultrasound analyzes. It was observed that the lowest serum activities of ALT and serum concentrations of total bilirubin occurred in the groups treated with the dry extract of Bidens pilosa, while only decreased serum concentrations of total bilirubin occurred in the group treated with Silymarin. Best liver recovery was also observed for the dry extract of B. pilosa at a 400mg/Kg dose by ultrasonography. This study showed that the dry extract of Bidens pilosa acted more efficiently in the treatment of acute toxic hepatitis induced in dogs than Silymarin.(AU)

Bidens pilosa L. é uma planta medicinal utilizada popularmente para tratamento de doenças hepáticas. Neste trabalho, o extrato seco das partes aéreas da Bidens pilosa e a silimarina, um fitocomplexo obtido dos frutos da Silybum marianum e comercializado como hepatoprotetor, foram testados em cães intoxicados experimentalmente de forma aguda com tetracloreto de carbono. A atividade hepática foi avaliada por meio dos perfis hematológico e bioquímico, análises histológica e ultrassonográfica. Observou-se que, nos grupos tratados com o extrato seco da Bidens pilosa, ocorreram as menores atividades séricas da ALT e de concentrações séricas de bilirrubina total, enquanto no grupo tratado com silimarina, ocorreu apenas diminuição de concentrações séricas de bilirrubina total. Melhor recuperação hepática também foi verificada para o extrato seco de B. pilosa na dose de 400mg/kg por ultrassonografia. Este estudo evidenciou que o extrato seco da Bidens pilosa atuou de forma mais eficiente no tratamento da hepatite aguda tóxica induzida em cães do que a silimarina.(AU)

A novel polyherbal formulation containing thymoquinone attenuates carbon tetrachloride-induced hepatorenal injury in a rat model

Objective: To evaluate a novel polyherbal formulation (BSVT) containing the standardized extracts from the leaves of Boerhavia diffusa, Solidago virgaurea, Vitex negundo, and thymoquinone in CCl

Resveratrol downregulates TGF-β1 and Smad3 expression and attenuates oxidative stress in CCl

Objective: To evaluate the effect of resveratrol against CCl

Resveratrol downregulates TGF-β1 and Smad3 expression and attenuates oxidative stress in CCl4-induced kidney damage in rats

Objective: To evaluate the effect of resveratrol against CCl4-induced nephrotoxicity. Methods: Forty-two male Wistar rats were divided into seven groups randomly. After six weeks, kidney weight, body weight, blood urea, serum creatinine, oxidative stress markers, and gene expression of renal transforming growth factor-beta1 (TGF-β1), TGF-β receptor type 1 (TGF-βR1) and Smad3 were determined. In addition, the protein level of TGF-β1 in the tissue lysate was measured. Results: Resveratrol had a protective role in renal tissue by the improvement of antioxidant balance and reduction of renal parameters such as creatinine and urea (P<0.001). In addition, the renal mRNA level of TGF-β1, TGF-βR1, Smad3, as well as the protein level of TGF-β1 were decreased in rats treated with resveratrol (P<0.001), compared to the CCl4 group. Conclusions: Overall, resveratrol shows a protective effect against nephrotoxicity in CCl4 treated rats by reducing oxidative stress status and modulating the TGF-β signaling.

Effects of silymarin on liver injury and bone metabolism induced by carbon tetrachloride in mice / 中国组织工程研究

BACKGROUND: Studies have found that silymarin has a regulatory role in multiple genes, which contributes to bone remodeling and prevents bone loss. In a mouse model of fracture healing, silymarin supplementation can improve tibia healing by increasing bone mineral density and serum alkaline phosphatase and osteocalcin levels. OBJECTIVE: To study the effect of silymarin on liver injury and bone metabolism induced by carbon tetrachloride in mice. METHODS: Twenty-four common Kunming mice, 10 weeks old, half male and half female, were randomly divided into three groups according to body mass. In the control group, subcutaneous injection of peanut oil 10 μL/g (double first dose) was given once every 5 days, followed by intragastric administration of 10 mL/kg/d distilled water. In the model group, animal models were made by subcutaneous injection of 40% carbon tetrachloride, followed by the same treatments as described in the control group. In the silymarin group, intragastric administration of silymarin solution 50 mg/kg/d was given after modeling. Treatments in each group lasted for 4 weeks. Each mouse was weighed every other week and was fasted for 12 hours the night before the final treatment. Under anesthesia, the mouse eyeballs were taken and blood sample from each mouse was taken to determine the serum aspartate aminotransferase and alanine aminotransferase activity; the liver was taken to measure the levels of malondialdehyde, glutathione peroxidase and superoxide dismutase in the liver homogenate; the right femur was taken to measure the bone calcium content; and the right tibia was taken for Micro CT detection to detect the changes in bone structure parameters. An approval by the Animal Ethics Committee of Guangdong Medical University was obtained with an approval No. PJ2013011. RESULTS AND CONCLUSION: Compared with the control group, the activity of aspartate aminotransferase and alanine aminotransferase in the model group was significantly increased (P < 0.05), glutathione peroxidase activity was significantly reduced (P < 0.05), bone calcium and tibial bone volume fraction, bone mineral density, and connection density were significantly reduced (P < 0.05), structural model index and anisotropy degree were significantly increased (P < 0.05), and significant liver damage and decreased bone mass and bone microstructure damage were observed. Compared with the model group, silymarin significantly reduced the activity of alanine aminotransferase (P < 0.05), and also significantly reduced the structural model index and the degree of anisotropy (P < 0.05), making the trabecular bone structure and trend more consistent. There was a clear network structure, and the bone microstructure remained intact. After the administration of carbon tetrachloride, the mice suffered liver damage with decreased bone mass and damaged bone microstructure, and silymarin administration had a certain preventive effect on liver damage and bone loss caused by carbon tetrachloride in mice.

Therapeutic effect of human adipose stem cells derived exosomes on carbon tetrachloride induced liver fibrosis in rats / 中国组织工程研究

BACKGROUND: Liver fibrosis has higher morbidity and mortality. Activation and proliferation of hepatic stellate cells is a key link in the progression of liver fibrosis. At present, there are still no effective anti-fibrosis agents targeting single links or targets. OBJECTIVE: To analyze the effect of human adipose stem cells derived exosomes on rats with liver fibrosis induced by carbon tetrachloride. METHODS: Human adipose stem cells were obtained from healthy people by enzyme dissolution method. After in vitro culture, human adipose stem cells derived exosomes were obtained by multiple ultrafiltration. Different concentrations of exosomes were used to treat the hepatic stellate cells activated by transforming growth factor β1. The human adipose stem cells activated by transforming growth factor β1 were treated with different concentrations of exosomes. The expression of α-smooth actin in the cells was detected by quantitative PCR, and the growth and apoptosis of activated hepatic stellate cells were detected by CCK-8 and flow cytometry respectively. Rat models of liver fibrosis were established by intraperitoneal injection of carbon tetrachloride and treated by tail vein injection of exosomes. Rat liver function, serum levels of type III procollagen and type IV collagen, and Ishak score were determined. Semi-quantitative analysis of liver fibrosis was performed. The expression levels of tissue inhibitor of matrix metalloproteinase-1, matrix metalloproteinase 9 and α-smooth actin in liver tissue were measured by immunofluorescence method. The study protocol was approved by the Animal Ethics Committee and Medical Ethics Committee, Tongji University, China in January, 2017. RESULTS AND CONCLUSION: Human adipose stem cells derived exosomes inhibited the proliferation of activated hepatic stellate cells. The possible mechanism is to inhibit the proliferation of activated macrophages, reduce the production of collagen fibers, α-smooth actin actin, and tissue inhibitor of matrix metalloproteinase-1, and to increase the expression of matrix metalloproteinase 9. These findings suggest that exosomes can be used to treat carbon tetrachloride induced liver fibrosis.

Protective effect of notoginsenoside R1 on carbon tetrachloride-induced liver fibrosis in rats / 中国组织工程研究

BACKGROUND: Previous studies have found that panax notoginseng saponins have a certain protective effect on immunological liver injury in mice. OBJECTIVE: To explore the therapeutic effect of notoginsenoside R1 on carbon tetrachloride-induced liver fibrosis in rats. METHODS: Experimental liver fibrosis model was made by carbon tetrachloride in male Sprague-Dawley rats. Then 30 g/L notoginsenoside R1 (60 mg/kg) was given once daily for 4 and 6 weeks in the treatment group. Rats in the control and model group were given distilled water of the same volume. Histopathological observation with hematoxylin-eosin staining and Masson’s trichrome staining was used to evaluate the changes of liver structure and fibrosis degree. The expression of collage type I, α-smooth muscle actin and transforming growth factor-β1 mRNA of hepatic tissue was measured by qRT-PCR method. The experimental protocol was approved by the Animal Experiment Ethics Committee of Kunming Medical University (approval No. KMMU2018018). RESULTS AND CONCLUSION: Liver histopathology showed that notoginsenoside R1 improved the degree of liver fibrosis. The expression levels of collagen type I, α-smooth muscle actin and transforming growth factor-β1 mRNA were reduced significantly in the treatment group compared with the model group (P < 0.05). But there was no significant difference after 4 and 6 weeks of treatment with notoginsenoside R1. Overall findings indicate that notoginsenoside R1 can slow down the progression of carbon tetrachloride-induced liver fibrosis in rats to a certain extent.

Effects of total flavonoids from Nymphaea candida on hepatic fibrosis induced by carbon tetrachloride in rats / 中草药

Objective: To investigate the preventive effect of total flavonoids from Nymphaea candida (NCTF) on carbon tetrachloride (CCl4)-induced hepatic fibrosis in rats. Methods: Wistar rats were randomly divided into control group, model group, NCTF groups (50, 100, 200 mg/kg) and colchicine group (0.2 mg/kg). Rats were subcutaneously injected with 50% CCl4 peanut oil solution (0.1 mL/100g, twice a week for 12 weeks) to induce hepatic fibrosis. In addition to control and model group given with 0.5% CMC-Na, the other groups were intragastrically administered with drugs (1.0 mL/100 g, once a day for 12 weeks), all rats were put to death. Blood, hepatic and splenic tissue were collected to detect liver indexes. Pathological histology observation by Masson and HE staining were performed for other hepatic tissues. The expression of α-smooth muscle actin (α-SMA) in hepatic tissue was analyzed by immunohistochemical technique. The levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total protein (TP) and albumin (ALB), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH) and nitric oxide (NO) were determined by automatic biochemical analyzer. The levels of serum laminin (LN), type III procollagen (PCIII), type IV collagen (CIV), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) were measured by enzyme-linked immunoassay assay (ELISA). Serum hyaluronic acid (HA) was detected by radioimmunoassay. Hepatic tissue hydroxyproline (Hyp) was measured by spectrophotometric method. Result Compared with the control group, the rats showed energielos, liver and spleen index of those were increased, as well as inflammatory cell infiltration and obvious fibrosis of hepatic tissue were appeared in the model group; The levels of ALT, AST, ALP, HA, LN, PCIII, CIV, MDA, NO, TNF-α, IL-1β and IL-6 in serum in model group were increased significantly (P < 0.05, 0.01); The levels of TP, ALB, GSH and SOD in serum were decreased significantly (P < 0.01); The expressions of α-SMA and Hyp in hepatic tissue were significantly improved (P < 0.01). Compared with the model group, the levels of ALT, AST, ALP, MDA, NO in serum in NCTF group were significantly reduced (P < 0.05, 0.01); The levels of HA, LN, PC III, CIV, TNF-α, IL-1β and IL-6 were significantly reduced (P < 0.05, 0.01); The levels of TP, ALB, GSH and SOD were significantly increased (P < 0.05, 0.01); The expressions of α-SMA and Hyp in hepatic tissue were significantly reduced (P < 0.05, 0.01). Compared with the model group, the pathological damage of hepatic tissue was significantly improved, the degeneration and necrosis of liver cells in rat were significantly reduced (P < 0.05). Conclusion: NCTF has a better anti-hepatic fibrosis effect, its mechanism is related to antioxidant, regulation of collagen synthesis and inhibition of the proinflammatory factors expression.