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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.
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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.
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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.
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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.
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Objective • To construct a non-alcoholic steatohepatitis (NASH) mouse model by the combination of Western diet (WD) and low-dose carbon tetrachloride (CCl4), and explore the time nodes of typical NASH pathological changes. Methods • Male 8-week C57BL/6 mice were fed WD and intraperitoneally injected with CCl4 at a dose of 2 μL/g of body weight per week to construct NASH models. At different time points, the fasting blood glucose, and the levels of triacylglyceride, glutamic-pyruvic transaminase and glutamic-oxaloacetic transaminase were tested; glucose tolerance was tested at the 24th week. Besides, the liver index was calculated and oil red O staining, Sirius red staining, hematoxylin-eosin staining and TUNEL test were conducted to evaluate liver pathological changes after liver sampling. Results • Between the control group and model group, there was no significant difference in fasting blood glucose and glucose tolerance test result, while the significant differences of liver index were observed at the 8th, 12th and 24th week (P<0.05). And at the 24th week, the levels of triacylglyceride, glutamic-pyruvic transaminase and glutamic-oxaloacetic transaminase were higher in the model group than those in the control group (P<0.05). According to the results of oil red O staining, Sirius red staining, hematoxylin-eosin staining and TUNEL test, in the model group, a large amount of small lipid droplets accumulation in the liver tissues was detected and hepatocytes were mainly in apoptotic state at the 8th week; large lipid droplets, hepatocellular ballooning and spot-like necrosis were observed, and hepatocyte apoptosis persisted at the 16th week; stage 3 fibrosis of liver was observed, and the number of spot-like necrosis increased but lipid droplets decreased, while hepatocytes were mainly in a proliferative state at the 24th week. Conclusion • The mouse model of NASH can be established successfully by WD combined with low-dose CCl4, which can simulate the pathologic features of NASH in a short time.
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Objective:To replicate the animal model of liver injury in rats by using carbon tetrachloride (CCl4), investigate the dynamic changes of early biomarkers of liver injury, namely glutamate dehydrogenase (GLDH), purine nucleotide phosphorylase(PNP), α-dynamic changes of glutathione-S-transferase (α-GST) and arginase 1(Arg1), and provide experimental evidence for early detection of acute liver injury. Method:Forty-eight Wistar rats were randomly divided into a blank group and a model group. The model group was intraperitoneally injected with 10 mL·kg-1 10% CCl4 olive oil solution, fasting but except water. Animals were sacrificed at 3, 6, 12, and 24 h. The serum liver function alanine aminotransferase(ALT), aspartate aminotransferase (AST), bilirubin (TBIL), alkaline phosphatase (ALP) levels, α-GST, Arg1, GLDH, PNP levels, and liver homogenate superoxide dismutase (SOD), glutathione (GSH), malondialdehyde (MDA) levels were then detected. Result:As compared with blank group, the levels of ALT, AST, TBIL, α-GST, Arg1, GLDH, PNP and MDA were increased significantly 3 h after administration, and SOD was decreased significantly(Pα-GST, ARG-, GLDH, TBIL, ALP and MDA were increased significantly, while GSH and SOD were decreased significantly (PPα-GST, Arg1, TBIL, ALP and MDA were significantly increased, while GSH and SOD were significantly decreased (PConclusion:α-GST, Arg1, GLDH and PNP have better sensitivity than traditional liver function test indicators, and can be used for early detection of liver injury induced by CCl4 in rats.
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OBJECTIVE: To investigate the effect of Rhaponticum uniflorum water extract (RUWE) on the oxidative stress and DNA damage of liver with an acute liver injury induced by carbon tetrachloride (CCl4) in mice. METHODS: Mice were randomly assigned to the normal control, model, bifendate (BFD), as well as high and low dose groups of RUWE. Animals were treated once daily for 7 d. At the end of the experiment, CCl4 was given intraperitoneally to the mice of groups, then the alanine aminotransferase (ALT), aspartate aminotransferase (AST), lipid hydroperoxide (LOOH), malondialdehyde (MDA), glutathione (GSH), catalase (CAT), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), Na+-K+-ATPase and Ca2+-Mg2+-ATPase were detected with the colorimetric method, and DNA damage was analyzed using the electrophoretic method. RESULTS: The administration with RUWE reduced the serum activities of ALT and AST, decreased the LOOH and MDA contents of liver, increased the CAT, GSH-Px, total SOD, Mn-SOD activities and GSH level of liver, increased the Na+-K+-ATPase and Ca2+-Mg2+-ATPase activities of liver mitochondria, and reduced the DNA damage of hepatocyte in mice with acute liver injury. CONCLUSION: RUWE has protective effect on acute liver injury induced by CCl4 in mice, probably via reduction of the oxidative stress and DNA damage of liver tissue.
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The effects of a fermentation filtrate of Ganoderma lucidum (FGL) on carbon tetrachloride (CCl4)-induced hepatic fibrosis were investigated in rats. Male Sprague-Dawley rats were orally administered with FGL (20 or 100 mg/kg) for 33 days, and orally administered with CCl4 (1.0 mL/kg; 2 mL/kg of 50% in corn oil) at 3-day intervals 1 h after FGL treatment. Body and liver weights, blood and histopathological findings in accordance with hydroxyproline concentrations were analyzed. Chronic exposure to CCl4 reduced the body weight gain, but increased liver weights and fibrosis, resulting in 3.35-fold increase in hydroxyproline level. Although FGL did not significantly reduce the CCl4-induced body and liver weight changes, it attenuated the increases in the hepatic fibrosis and hydroxyproline contents. Taken together, it is suggested that FGL might prevent hepatic fibrosis, and that FGL or its ingredient could be a potential candidate for the prevention of chronic hepatic disorders.
Subject(s)
Animals , Humans , Male , Rats , Body Weight , Carbon Tetrachloride , Fermentation , Fibrosis , Ganoderma , Hydroxyproline , Liver , Rats, Sprague-Dawley , Reishi , Weights and Measures , Zea maysABSTRACT
Objective: To study the protective function of Gardenia yellow (GY) against CCl 4 induced hepatotoxicity in mice.Methods: Healthy Kunming male mice, weighting (20?2) g, 10 per group were randomized into 5 groups:control group, CCl 4 injured group, low dose group(CCl 4 injured+0.1 ml GY solution), medium dose group(CCl 4 injured+ 0.2 ml GY solution) and high dose group(CCl 4 injured+0.4ml GY solution). GY solution was given i.g. 5 d prior to CCl 4 injury. Serum glutamate pyruvate transaminase(SGPT) and glutamic oxaloacetic transaminase(SGOT) and lactic dehydrogenase(LDH) activities were determined 18 h after CCl 4 injury. Hepatic malondialdehyde(MDA), glutathione(GSH) and liver index were also detected. Results: In GY treated groups, the increases of serum SGOT, SGPT, LDH activities and liver GSH were inhibited obviously. The elevations of MDA and liver index were prevented significantly. The lesions in liver lobule were ameliorated obviously. Conclusion: Gardenia yellow can protect against CCl 4 induced hepatotoxicity.