AIM:To evaluate the hepatoprotective roles of (Z)5-(4-methoxybenzylidene)thiazolidine-2,4-dione (SKLB010) against carbon tetrachloride (CCl 4)-induced acute and chronic liver injury and its underlying mechanisms of ac...AIM:To evaluate the hepatoprotective roles of (Z)5-(4-methoxybenzylidene)thiazolidine-2,4-dione (SKLB010) against carbon tetrachloride (CCl 4)-induced acute and chronic liver injury and its underlying mechanisms of action.METHODS:In the first experiment,rats were weighed and randomly divided into 5 groups (five rats in each group) to assess the protective effect of SKLB010 on acute liver injury.For induction of acute injury,rats were administered a single intraperitoneal injection of 2 mL/kg of 50% (v/v) CCl 4 dissolved in olive oil (1:1).Group 1 was untreated and served as the control group;group 2 received CCl 4 for induction of liver injury and served as the model group.In groups 3,4 and 5,rats receiving CCl 4 were also treated with SKLB010 at doses of 25,50 and 100 mg/kg,respectively.Blood samples were collected at 6,12 and 24 h after CCl 4 intoxication to determine the serum activity of alanine amino transferase.Tumour necrosis factor-α (TNF-α),interleukin-1β (IL-1β) were determined using enzyme-linked immunosorbent assay.At 24 h after CCl 4 injection,liver fibrogenesis was evaluated by hematoxylin-eosin (HE) staining and immunohistochemical analyses.Cytokine transcript levels of TNF-α,IL-1β and inducible nitric oxide synthase in the liver tissues of rats were measured using a reverse transcriptase reverse transcription-polymerase chain reaction technique.In the second experiment,rats were randomly divided into 2 groups (15 rats in each group),and liver injury in the CCl 4-administered groups was induced by a single intraperitoneal injection of 2 mL/kg of 50% (v/v) CCl 4 dissolved in olive oil (1:1).The SKLB010-treated groups received oral 100 mg/kg SKLB010 before CCl 4 administration.Five rats in each group were sacrificed at 2 h,6 h,12 h after CCl 4 intoxication and small fortions of livers were rapidly frozen for extraction of total RNA,hepatic proteins and glutathione (GSH) assays.In the hepatic fibrosis model group,rats were randomly divided into 2 groups (5 rats each group).Rats were injected intraperitoneally with a mixture of CCl 4 (1 mL/kg body weight) and olive oil [1:1 (v/v)] twice a week for 4 wk.In the SKLB010-treated groups,SKLB010 (100 mg/kg) was given once daily by oral gavage for 4 wk after CCl 4 administration.The rats were sacrificed one week after the last injection and the livers from each group were harvested and fixed in 10% formalin for HE and immunohistochemical staining.RESULTS:In this rat acute liver injury model,oral administration of SKLB010 blocked liver tissue injury by down-regulating the serum levels of alanine aminotransferase,suppressing inflammatory infiltration to liver tissue,and improving the histological architecture of liver.SKLB010 inhibited the activation of NF-κB by suppressing the degradation of IκB,and prevented the secretion of pro-inflammatory mediators such as tumor necrosis factor-α,interleukin-1β,and the reactive free radical,nitric oxide,at the transcriptional and translational levels.In this chronic liver fibrosis model,treatment with 100 mg/kg per day SKLB010 attenuated the degree of hepatic fibrosis and area of collagen,and blocked the accumulation of smooth-muscle actinexpressed cells.CONCLUSION:These results suggest that SKLB010 is a potent therapeutic agent for the treatment of CCl 4 induced hepatic injury.展开更多
[Objectives] To study the protection of compatibility of Saikosapon d and Baicalin on carbon tetrachloride( CCl_4) injured L-02 cells. [Methods] Normal human hepatocyte cell line L-02 cells were cultured in vitro,and ...[Objectives] To study the protection of compatibility of Saikosapon d and Baicalin on carbon tetrachloride( CCl_4) injured L-02 cells. [Methods] Normal human hepatocyte cell line L-02 cells were cultured in vitro,and CCl_4 was used to induce hepatocellular injury. Interventions were carried out with Saikosaponin d and Baicalin at different dosage. The proliferation of L-02 cells in each group was determined by methylthiazolyl tetrazolium( MTT) assay; the levels of AST and ALT in the culture supernatants were detected by enzyme-linked immunosorbent assay( ELISA); the expressions of TLR4 and NFκBp65 proteins in each group were determined by immunohistochemistry.[Results] In the CCl_4 injured group,the proliferation of L-02 cells was significantly declined,the levels of AST and ALT in cell culture medium were significantly increased,and the expressions of TLR4 and NFκBp65 in L-02 cells were increased; after the intervention of Saikosaponin d and Baicalin,1. 75 μg/mL group and 1. 5 μg/mL group had an effect of promoting the proliferation of L-02 cells and could reduce the levels of AST and ALT in the cell culture medium,and TLR4 and NFκBp65 proteins in L-02 cells also had a certain inhibitory effect. [Conclusions] The compatibility of Saikosapon d and Baicalin has a certain protective effect on CCl_4 injured L-02 cells. The protection mechanism may be related with its down-regulating TLR4-NFκB signaling pathway and reducing the inflammation.展开更多
[Objectives]To study the protective effect of Ershiwuwei Songshi Pills on chronic liver injury induced by carbon tetrachloride(CCL4)in rats before and after the modification conforming to the compatibility theory of T...[Objectives]To study the protective effect of Ershiwuwei Songshi Pills on chronic liver injury induced by carbon tetrachloride(CCL4)in rats before and after the modification conforming to the compatibility theory of Tibetan medicine,and to explore its action mechanism.[Methods]Male Wistar rats were randomly divided into the blank control group,model group,Hugan tablets group(0.490 g/kg),Ershiwuwei Songshi Pills group(0.117 g/kg),and Modified Ershiwuwei Songshi Pills group(removing cinnabaris,Aristolochia contorta,and Aconitum naviculare,0.105 g/kg).Except the blank group,the remaining groups were injected subcutaneously with 20%carbon tetrachloride olive oil solution every 3 d,and modeled for 6 weeks.During this time,intragastrically administered corresponding drugs.Six weeks later,blood was taken from the femoral artery,and the rats were killed through dislocating the cervical spine,the liver was taken,and the content of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)was determined.Then,liver fibrosis indicators tumor necrosis factor-α(TNF-α),nuclear factor-κB(NF-κB),interleukin-6(IL-6)and interleukin-1β(IL-1β)were detected by immunohistochemical method.[Results]Compared with the model group,the pathological map of the liver section showed that liver injury was improved in each administration group.The serum ALT and AST contents in rats of each administration group were significantly reduced(P<0.05),and the protein expressions of NF-κB,TNF-α,IL-1βand IL-6 in liver tissue were also reduced by varying degrees(P<0.05).[Conclusions]Ershiwuwei Songshi Pills and its modification group have a protective effect on liver injury induced by carbon tetrachloride.The modified prescription conforms to the compatibility rules of Tibetan medicine.The mechanism may be related to reducing the damage caused by inflammatory factors through regulating the role of inflammatory signaling pathway.Thus,it can be used as a reference for future optimization proposals.展开更多
Background and aim:Advanced liver fibrosis is a major risk for developing hepatocellular carcinoma(HCC)in chronic hepatitis C virus(HCV)patients.Direct-acting antivirals(DAAs)which are used for treating HCV infection,...Background and aim:Advanced liver fibrosis is a major risk for developing hepatocellular carcinoma(HCC)in chronic hepatitis C virus(HCV)patients.Direct-acting antivirals(DAAs)which are used for treating HCV infection,produce more than 90%cure rate but do not seem to diminish the rate of occurrence or recurrence of HCC.This study aimed to investigate the effect of DAAs sofosbuvir(SOF)and daclatasvir(DAC)on carbon tetrachloride(CCl4)-induced fibrotic changes in mice.Methods:Eighty adult male Swiss albino mice were randomly allocated into 8 groups(10 mice/group):normal control group,SOF group(receiving SOF 80 mg/kg body weight(BW),oral gavage,daily),DAC group(receiving DAC 30 mg/kg BW,oral gavage,daily),SOF t DAC group(receiving a combination of both,daily),CCl4 model group(receiving CCl42 mL/kg BW,intraperitoneal twice weekly)and three CCl4-intoxicated groups receiving either SOF or DAC or their combination.All CCl4 groups received CCl4 for 12 weeks followed by DAAs for another 12 weeks.Results:CCl4-induced a significant elevation of alanine aminotransferase(ALT)and aspartate aminotransferase(AST),and produced histopathological evidence of fibrosis and liver degeneration along with a significant increase(P0.001)of the proliferation markers(proliferating cell nuclear antigen(PCNA)and Ki-67),hepatic stellate cells(HSCs)activation markers(alpha-smooth muscle actin(a-SMA)and glial fibrillary acidic protein(GFAP)),fibrosis marker(matrix metalloproteinase-9(MMP-9))and proinflammatory cytokine(tumor necrosis factor-alpha(TNF-a)).CCl4-intoxicated mice treated with SOF,DAC,or their combination revealed a significant amelioration(P0.001)of CCl4-induced elevation of liver enzymes,fibrotic changes,and liver degeneration along with a significant attenuation(P0.001)of CCl4-induced upregulation of all tested markers.The effects of SOF,DAC,and their combination on liver enzymes were comparable while the effect of SOF t DAC combination on mitigating CCl4-induced upregulation of the proliferation and HSCs activation markers was significantly stronger than either SOF or DAC alone.As for MMP-9 and TNF-a,the effects of DAC and SOF t DAC combination were comparable and both were more significant than that of SOF alone.Conclusions:SOF and DAC may possess an antifibrotic effect that is independent of their role as antiviral agents against CCl4-induced liver injury.This might exclude the role of DAAs in early occurrence or accelerated recurrence of HCC through the progression of the HCV patients'pre-existing fibrosis.However,HCC patients treated with DAAs should be closely monitored with continuous HCC surveillance during and post-therapy.展开更多
基金Supported by National Natural Science Foundation of China and National Key Technologies R and D Program of the 11th five-year plan,No.2009ZX09501-015
文摘AIM:To evaluate the hepatoprotective roles of (Z)5-(4-methoxybenzylidene)thiazolidine-2,4-dione (SKLB010) against carbon tetrachloride (CCl 4)-induced acute and chronic liver injury and its underlying mechanisms of action.METHODS:In the first experiment,rats were weighed and randomly divided into 5 groups (five rats in each group) to assess the protective effect of SKLB010 on acute liver injury.For induction of acute injury,rats were administered a single intraperitoneal injection of 2 mL/kg of 50% (v/v) CCl 4 dissolved in olive oil (1:1).Group 1 was untreated and served as the control group;group 2 received CCl 4 for induction of liver injury and served as the model group.In groups 3,4 and 5,rats receiving CCl 4 were also treated with SKLB010 at doses of 25,50 and 100 mg/kg,respectively.Blood samples were collected at 6,12 and 24 h after CCl 4 intoxication to determine the serum activity of alanine amino transferase.Tumour necrosis factor-α (TNF-α),interleukin-1β (IL-1β) were determined using enzyme-linked immunosorbent assay.At 24 h after CCl 4 injection,liver fibrogenesis was evaluated by hematoxylin-eosin (HE) staining and immunohistochemical analyses.Cytokine transcript levels of TNF-α,IL-1β and inducible nitric oxide synthase in the liver tissues of rats were measured using a reverse transcriptase reverse transcription-polymerase chain reaction technique.In the second experiment,rats were randomly divided into 2 groups (15 rats in each group),and liver injury in the CCl 4-administered groups was induced by a single intraperitoneal injection of 2 mL/kg of 50% (v/v) CCl 4 dissolved in olive oil (1:1).The SKLB010-treated groups received oral 100 mg/kg SKLB010 before CCl 4 administration.Five rats in each group were sacrificed at 2 h,6 h,12 h after CCl 4 intoxication and small fortions of livers were rapidly frozen for extraction of total RNA,hepatic proteins and glutathione (GSH) assays.In the hepatic fibrosis model group,rats were randomly divided into 2 groups (5 rats each group).Rats were injected intraperitoneally with a mixture of CCl 4 (1 mL/kg body weight) and olive oil [1:1 (v/v)] twice a week for 4 wk.In the SKLB010-treated groups,SKLB010 (100 mg/kg) was given once daily by oral gavage for 4 wk after CCl 4 administration.The rats were sacrificed one week after the last injection and the livers from each group were harvested and fixed in 10% formalin for HE and immunohistochemical staining.RESULTS:In this rat acute liver injury model,oral administration of SKLB010 blocked liver tissue injury by down-regulating the serum levels of alanine aminotransferase,suppressing inflammatory infiltration to liver tissue,and improving the histological architecture of liver.SKLB010 inhibited the activation of NF-κB by suppressing the degradation of IκB,and prevented the secretion of pro-inflammatory mediators such as tumor necrosis factor-α,interleukin-1β,and the reactive free radical,nitric oxide,at the transcriptional and translational levels.In this chronic liver fibrosis model,treatment with 100 mg/kg per day SKLB010 attenuated the degree of hepatic fibrosis and area of collagen,and blocked the accumulation of smooth-muscle actinexpressed cells.CONCLUSION:These results suggest that SKLB010 is a potent therapeutic agent for the treatment of CCl 4 induced hepatic injury.
基金Supported by Project of Shaanxi Administration of Traditional Chinese Medicine(zy06)Special Project of Education Department of Shaanxi Provincial Government(12JK1016)Program of Shaanxi Provincial Department of Science and Technology(2013jk4023)
文摘[Objectives] To study the protection of compatibility of Saikosapon d and Baicalin on carbon tetrachloride( CCl_4) injured L-02 cells. [Methods] Normal human hepatocyte cell line L-02 cells were cultured in vitro,and CCl_4 was used to induce hepatocellular injury. Interventions were carried out with Saikosaponin d and Baicalin at different dosage. The proliferation of L-02 cells in each group was determined by methylthiazolyl tetrazolium( MTT) assay; the levels of AST and ALT in the culture supernatants were detected by enzyme-linked immunosorbent assay( ELISA); the expressions of TLR4 and NFκBp65 proteins in each group were determined by immunohistochemistry.[Results] In the CCl_4 injured group,the proliferation of L-02 cells was significantly declined,the levels of AST and ALT in cell culture medium were significantly increased,and the expressions of TLR4 and NFκBp65 in L-02 cells were increased; after the intervention of Saikosaponin d and Baicalin,1. 75 μg/mL group and 1. 5 μg/mL group had an effect of promoting the proliferation of L-02 cells and could reduce the levels of AST and ALT in the cell culture medium,and TLR4 and NFκBp65 proteins in L-02 cells also had a certain inhibitory effect. [Conclusions] The compatibility of Saikosapon d and Baicalin has a certain protective effect on CCl_4 injured L-02 cells. The protection mechanism may be related with its down-regulating TLR4-NFκB signaling pathway and reducing the inflammation.
基金Supported by the Fundamental Research Funds for the Central Universities(2018NZD19)Systematic Study and Industrial Demonstration of Prevention and Treatment of Liver Diseases with Tibetan Medicines of the Qinghai-Tibet PlateauInnovating Research Program of Postgraduates of Southwest Minzu University in 2018(CX2018SZ81).
文摘[Objectives]To study the protective effect of Ershiwuwei Songshi Pills on chronic liver injury induced by carbon tetrachloride(CCL4)in rats before and after the modification conforming to the compatibility theory of Tibetan medicine,and to explore its action mechanism.[Methods]Male Wistar rats were randomly divided into the blank control group,model group,Hugan tablets group(0.490 g/kg),Ershiwuwei Songshi Pills group(0.117 g/kg),and Modified Ershiwuwei Songshi Pills group(removing cinnabaris,Aristolochia contorta,and Aconitum naviculare,0.105 g/kg).Except the blank group,the remaining groups were injected subcutaneously with 20%carbon tetrachloride olive oil solution every 3 d,and modeled for 6 weeks.During this time,intragastrically administered corresponding drugs.Six weeks later,blood was taken from the femoral artery,and the rats were killed through dislocating the cervical spine,the liver was taken,and the content of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)was determined.Then,liver fibrosis indicators tumor necrosis factor-α(TNF-α),nuclear factor-κB(NF-κB),interleukin-6(IL-6)and interleukin-1β(IL-1β)were detected by immunohistochemical method.[Results]Compared with the model group,the pathological map of the liver section showed that liver injury was improved in each administration group.The serum ALT and AST contents in rats of each administration group were significantly reduced(P<0.05),and the protein expressions of NF-κB,TNF-α,IL-1βand IL-6 in liver tissue were also reduced by varying degrees(P<0.05).[Conclusions]Ershiwuwei Songshi Pills and its modification group have a protective effect on liver injury induced by carbon tetrachloride.The modified prescription conforms to the compatibility rules of Tibetan medicine.The mechanism may be related to reducing the damage caused by inflammatory factors through regulating the role of inflammatory signaling pathway.Thus,it can be used as a reference for future optimization proposals.
文摘Background and aim:Advanced liver fibrosis is a major risk for developing hepatocellular carcinoma(HCC)in chronic hepatitis C virus(HCV)patients.Direct-acting antivirals(DAAs)which are used for treating HCV infection,produce more than 90%cure rate but do not seem to diminish the rate of occurrence or recurrence of HCC.This study aimed to investigate the effect of DAAs sofosbuvir(SOF)and daclatasvir(DAC)on carbon tetrachloride(CCl4)-induced fibrotic changes in mice.Methods:Eighty adult male Swiss albino mice were randomly allocated into 8 groups(10 mice/group):normal control group,SOF group(receiving SOF 80 mg/kg body weight(BW),oral gavage,daily),DAC group(receiving DAC 30 mg/kg BW,oral gavage,daily),SOF t DAC group(receiving a combination of both,daily),CCl4 model group(receiving CCl42 mL/kg BW,intraperitoneal twice weekly)and three CCl4-intoxicated groups receiving either SOF or DAC or their combination.All CCl4 groups received CCl4 for 12 weeks followed by DAAs for another 12 weeks.Results:CCl4-induced a significant elevation of alanine aminotransferase(ALT)and aspartate aminotransferase(AST),and produced histopathological evidence of fibrosis and liver degeneration along with a significant increase(P0.001)of the proliferation markers(proliferating cell nuclear antigen(PCNA)and Ki-67),hepatic stellate cells(HSCs)activation markers(alpha-smooth muscle actin(a-SMA)and glial fibrillary acidic protein(GFAP)),fibrosis marker(matrix metalloproteinase-9(MMP-9))and proinflammatory cytokine(tumor necrosis factor-alpha(TNF-a)).CCl4-intoxicated mice treated with SOF,DAC,or their combination revealed a significant amelioration(P0.001)of CCl4-induced elevation of liver enzymes,fibrotic changes,and liver degeneration along with a significant attenuation(P0.001)of CCl4-induced upregulation of all tested markers.The effects of SOF,DAC,and their combination on liver enzymes were comparable while the effect of SOF t DAC combination on mitigating CCl4-induced upregulation of the proliferation and HSCs activation markers was significantly stronger than either SOF or DAC alone.As for MMP-9 and TNF-a,the effects of DAC and SOF t DAC combination were comparable and both were more significant than that of SOF alone.Conclusions:SOF and DAC may possess an antifibrotic effect that is independent of their role as antiviral agents against CCl4-induced liver injury.This might exclude the role of DAAs in early occurrence or accelerated recurrence of HCC through the progression of the HCV patients'pre-existing fibrosis.However,HCC patients treated with DAAs should be closely monitored with continuous HCC surveillance during and post-therapy.
