AIM： To investigate if iron deposition and fat accumulation in the liver play a pathogenetic role in dimethylnitrosamine （DMN）-induced liver fibrosis in rat.METHODS： Thirty rats were treated with DMN at does conse...AIM： To investigate if iron deposition and fat accumulation in the liver play a pathogenetic role in dimethylnitrosamine （DMN）-induced liver fibrosis in rat.METHODS： Thirty rats were treated with DMN at does consecutive days of 10 μL/kg daily, i.p., for 3 consecutive day each week for 4 wk. Rats （n = 30） were sacrificed on the first day （model group A） and 21st d （model group B） after cessation of DMN injection. The control group （n = 10） received an equivalent amount of saline. Liver tissues were stained with hematoxylin ＆ eosin （HE） and Masson and Prussian blue assay and oberserved under electron microscopy. Serum alanine aminotransferase （ALT）and liver tissue hydroxyproline （Hyp） content were tested.RESULTS： The liver fibrosis did not automaticallyreverse, which was similar to previous reports, the perilobular deposition of iron accompanied with collagen showed marked characteristics at both the first and 21st d after cessation of DMN injection. However, fat accumulation in hepatocytes occurred only at the 21^st d after cessation of DMN injection.CONCLUSION： Iron deposition and fat accumulation may play important roles in pathological changes in DMN-induced rat liver fibrosis. The detailed mechanisms of these characteristics need further research.展开更多
AIM： To evaluate the therapeutic effect of Chunggan extract （CGX）, a modified traditional Chinese hepatotherapeutic herbal, on the dimethylnitrosamine （DMN）-induced chronic liver injury model in rats. METHODS： L...AIM： To evaluate the therapeutic effect of Chunggan extract （CGX）, a modified traditional Chinese hepatotherapeutic herbal, on the dimethylnitrosamine （DMN）-induced chronic liver injury model in rats. METHODS： Liver injuries were induced in Wistar rats by injection of DMN （ip, 10 mg/mL per kg） for 3 consecutive days per week for 4 wk. The rats were administered with CGX Coo, 100 or 200 mg/kg per day） or distilled water as a control daily for 4 wk starting from the 15^th d of the DMN treatment. Biochemical parameters （serum albumin, bilirubin, ALP, AST and ALT）, lipid peroxides, hydroxyproline, as well as histological changes in liver tissues were analyzed. In addition, gene expression of TNF-α, TGF-β, TIMP-1, TIMP-2, PDGF-β, and MMP-2, all of which are known to be associated with liver fibrosis, were analyzed using real-time PCR. RESULTS： CGX administration restored the spleen weight to normal alter having been increased by DMN treatment. Biochemical analysis of the serum demonstrated that CGX significantly decreased the serum level of ALP （P 〈 0.05）, ALT （P 〈 0.02）, and AST （P 〈 0.02） that had been elevated by DMN treatment. CGX administration moderately lowered lipid peroxide production and markedly lowered hydroxyproline generation caused by DMN treatment in accordance with histopathological examination. DMN treatment induced a highly upregulated expression of TNF-α, TGF-β,TIMP-1, TIMP-2, PDGF-β, and MMP-2. Of these, the gene expression encoding PDGF-β and MMP-2 was still further enhanced 2 wk after secession of the 4-wk DMN treatment, and was remarkably ameliorated by CGX administration. CONCLUSION： CGX exhibits hepatotherapeutic properties against chronic hepatocellular destruction and consequential liver fibrosis.展开更多
Objective:To observe the expression level of TNF-α mRNA in rats with hepatic fibrosis induced by dimethylnitrosamine (DMN) and to explore its relationship with collagen metabolism and its diagnostic value for hepatic...Objective:To observe the expression level of TNF-α mRNA in rats with hepatic fibrosis induced by dimethylnitrosamine (DMN) and to explore its relationship with collagen metabolism and its diagnostic value for hepatic fibrosis.Methods: Twenty-five male Wistar rats were randomly divided into normal control group (n=10) and model group (n=15). Model rats were induced by DMN for 4 weeks and at final stage were executed. TNF-α mRNA were detected by RT-PCR and the inflammatory necrosis and collagen deposition in hepatic tissue were observed by HE stain and Sirius red stain. The liver functions were determined by automatic biochemical analytic device. The serum marks of liver fibrosis, such as HA, LN and Ⅳ-C were measured with ELISA and RIA. Results: In this study, the rat model of liver fibrosis induced by DMN was successfully constructed. RT-PCR reveals that TNF-α mRNA expression in control group is lower than that of model group. The liver functions of model group were impaired compared with those of the control group (P<0.01). Semi-quantitive analysis revealed that TNF-α/β-actin of normal rats was 0.39±0.12, while 0.93±0.05 of model rats. The concentration of HA (434.44±98.81 vs 252.9±26.59 ng/ml, P<0.01), LN (70.67±6.32 vs 37.90±5.97 ng/ml, P<0.01) and Ⅳ-C (79.39±10.52 vs 21.40±4.17 ng/ml, P<0.01) were significantly increased in the model group as well. Changes of the indexes were similar to the pathological damage of the liver. Conclusion: The results suggested that activation of TNF-α in liver tissues may be the common pathogenic mechanism of liver fibrosis. TNF-α may be a useful index for the diagnosis of hepatic fibrosis which worthies further investigation.展开更多
Salvia miltiorrhiza （Sm） is a traditional herbal medicine with multiple effects on various diseases. Its water-soluble parts have been used to produce injectional powder. In this study, liver fibrosis rats were indu...Salvia miltiorrhiza （Sm） is a traditional herbal medicine with multiple effects on various diseases. Its water-soluble parts have been used to produce injectional powder. In this study, liver fibrosis rats were induced by intraperitoneal injection of dimethylnitrosarnine for 3 consecutive days per week for 4 weeks. After 2 weeks, rats in the positive drug group were subcutaneously injected with 8×10^5 IU/kg IFNα2b, while the Sm treatment groups were intraperitoneally injected with 50, 100 and 200 mg/kg solution of Sm injectional powder, respectively, for 6 days per week for 4 weeks. The results showed that either IFNα2b or the Sm injectional powder significantly increased the body weight and liver to spleen ratio, and three doses of the powder brought down the spleen index. Serum analysis showed that both IFNα2b and the Sm powder reduced levels of alanine transaminase and total bilirubin, while only 100 and 200 mg/kg of the Sm powder ameliorated aspartate transaminase and albumin levels. In the collagen examination, reduced hyaluronic acid and procollagen type III levels, less fibrous hyperplasia and collagen deposits, and improved hepatocyte states were clearly observed in rats treated with either IFNα2b or Sm injectional powder. In addition, the mechanism of action of the Sm powder was also studied. Immunohistochemical staining showed that IFNα2b and Sm injectional powder significantly down-regulated the expression of transforming growth factor-β1 （TGF-β1） and platelet derived growth factor （PDGF）. In conclusion, Sm injectional powder has protective effects on dimethylnitrosamine-initiated liver fibrosis in rats, and the mechanism may include the down-regulation of TGF-β1 and PDGF.展开更多
Objective：To study the effect of total flavonoids of Astmgali Radix（TFA）on liver cirrhosis induced with dimethylnitrosamine（DMN）in rats,and the effect on peroxisome proliferator-activated receptorγ（PPARγ）,unc...Objective：To study the effect of total flavonoids of Astmgali Radix（TFA）on liver cirrhosis induced with dimethylnitrosamine（DMN）in rats,and the effect on peroxisome proliferator-activated receptorγ（PPARγ）,uncoupling protein 2（UCP2）and farnesoid X receptor（FXR）.Methods：Fifty-three Sprague-Dawley rats were randomly divided into a control group（10 rats）and a DMN group（43 rats）.Rats in the DMN group were given DMN for 4 weeks and divided randomly into a model group（14 rats）,a low-dosage TFA group（14 rats）and a high-dosage TFA group（15 rats）in the 3rd week.Rats were given TFA for 4 weeks at the dosage of 15 and 30 mg/kg in the low-and high-TFA groups,respectively.At the end of the experiment blood and liver samples were collected.Serum liver function and liver tissue hydroxyproline content were determined.hematoxylin-eosin（HE）,Sirus red and immunohistochemical stainings of collagenⅠ,smooth muscle actin（α-SMA）was conducted in paraffinembedded liver tissue slices.Real time polymerase chain reaction（PCR）was adopted to determine PPARγ,UCP2 and FXR m RNA levels.Western blot was adopted to determine protein levels of collagenⅠ,α-SMA,PPARγ,UCP2 and FXR.Results：Compared with the model group,TFA increased the ratio of liver/body weight（low-TFA group P〈0.05,high-TFA group P〈0.01）,improved liver biochemical indices（P〈0.01 for ALT,AST,GGT in both groups,P〈0.05 for albumin and TBil in the high-TFA group）and reduced liver tissue hydroxproline content（P〈0.01 in both groups）in treatment groups significantly.HE staining showed that TFA alleviated liver pathological changes markedly and Sirus red staining showed that TFA reduced collagen deposition,alleviated formation and extent of liver pseudolobule.CollagenⅠandα-SMA immunohistochemical staining showed that staining area and extent markedly decreased in TFA groups compared with the model group.TFA could increase PPARγ,it regulated target UCP2,and FXR levels significantly compared with the model group（in the low-TFA group all P〈0.05,in the high group all P〈0.01）.Conclusions：TFA could improve liver function,alleviate liver pathological changes,and reduce collagen deposition and formation of liver pseudolobule in rats with liver cirrhosis.The antifibrotic effect of TFA was through regulating PPARγsignal pathway and the interaction with FXR.展开更多
AIM: To evaluate the antifibrotic effect of different doses of recombinant human Gamma-Interferon (IFN-gamma) in two rat models of hepatic fibrosis, and to observe its effect on moderate chronic hepatitis B virus fibr...AIM: To evaluate the antifibrotic effect of different doses of recombinant human Gamma-Interferon (IFN-gamma) in two rat models of hepatic fibrosis, and to observe its effect on moderate chronic hepatitis B virus fibrosis. METHODS: Hepatic fibrosis was successfully induced in 150 and 196 rats by subcutaneous injection of carbon tetrachloride (CCl4) and intraperitoneal injection of dimethylnitrosamine (DMN), respectively. Each of the two model groups was divided into: (1) fibrotic model group; (2) colchicine treatment group (0.1 mg/kg/day, gastrogavage for 8 weeks); (3) high-dose IFN-gamma group (15 MU/kg per day, i.m. for 8 weeks); (4) medium-dose IFN-gamma group (5 MU/kg daily, i.m. for 8 weeks); and (5) Y low-dose IFN-gamma group (1.67 MU/kg daily, i.m. for 8 weeks). Another group of 10 rats without any treatment was used as normal controls. At the end of the experiment, semi-quantitative histopathological scores of inflammation and fibrosis, liver alpha smooth muscle actin (alpha-SMA) expression level, liver hydroxyl proline content and serum hyaluronic acid levels were compared. And 47 medium chronic hepatitis B viral fibrosis patients were studied. They were given IFN-gamma treatment, 100 MU/day i.m. for the first three months and 100 MU qod i.m. for the next six months. Semi-quantitative pathological scores of inflammation and fibrosis and serum hepatic fibrosis indices were compared within the 9 months. RESULTS: In animal experiment, the pathological fibrosis scores and liver hydroxyl proline content were found to be significantly lower in rats treated with different doses of IFN-gamma as compared with rats in fibrotic model group induced by either CCl4 or DMN, in a dose-dependent manner. For CCl4-induced model, pathological fibrosis scores in high, medium and low doses IFN-gamma groups were 5.10 +/- 2.88, 7.70 +/- 3.53 and 8.00 +/- 3.30, respectively, but the score was 14.60 +/- 7.82 in fibrotic model group. Hydroxyl proline contents were 2.83 +/- 1.18, 3.59 +/- 1.22 and 4.80 +/- 1.62, in the three IFN-gamma groups, and 10.01 +/- 3.23 in fibrotic model group. The difference was statistically significant (P【0.01). Similar results were found in DMN-induced model. Pathological fibrosis scores were 6.30 +/- 0.48, 8.10 +/- 2.72 and 8.30 +/- 2.58, in high, medium and low doses IFN-gamma groups, and 12.60 +/- 3.57 in fibrotic model group. Hydroxyl proline contents were 2.72 +/- 0.58, 3.14 +/- 0.71 and 3.62 +/- 1.02, in the three IFN-gamma groups, and 12.79 +/- 1.54 in fibrotic model group. The difference was statistically significant (P【0.01).Serum hepatic fibrosis indices decreased significantly in the 47 patients after IFN-gamma treatment (HA: 433.38 +/- 373.00 vs 281.57 +/- 220.48; LN: 161.22 +/- 41.02 vs 146 +/- 35 +/- 44. 67; PC III: 192.59 +/- 89.95 vs 156.98 +/- 49.22; C-I: 156.30 +/- 44.01 vs 139.14 +/- 34.47) and the differences between the four indices were significant (P 【0.05). Thirty-three patients received two liver biopsies, one before and one after IFN-gamma treatment. In thirty of 33 patients IFN-gamma had better effects according to semi-quantitative pathological scores (8.40 +/- 5.83 vs 5.30 +/- 4.05, P【0.05). CONCLUSION: All the three doses of IFN-gamma are effective in treating rat liver fibrosis induced by either CCl4 or DMN, the higher the dose, the better the effect. And IFN-gamma is effective for patients with moderate chronic hepatitis B viral fibrosis.展开更多
AIM: To evaluate the efficacy of umbilical cord-derived mesenchymal stem cells(UC-MSCs) transplantation in the treatment of liver fibrosis.METHODS: Cultured human UC-MSCs were isolated and transfused into rats with li...AIM: To evaluate the efficacy of umbilical cord-derived mesenchymal stem cells(UC-MSCs) transplantation in the treatment of liver fibrosis.METHODS: Cultured human UC-MSCs were isolated and transfused into rats with liver fibrosis induced by dimethylnitrosamine(DMN). The effects of UC-MSCs transfusion on liver fibrosis were then evaluated by histopathology; serum interleukin(IL)-4 and IL-10 levels were also measured. Furthermore, Kupffer cells(KCs) in fibrotic livers were isolated and cultured to analyze their phenotype. Moreover, UC-MSCs were cocultured with KCs in vitro to assess the effects of UCMSCs on KCs' phenotype, and IL-4 and IL-10 levels were measured in cell culture supernatants. Finally, UCMSCs and KCs were cultured in the presence of IL-4 antibodies to block the effects of this cytokine, followed by phenotypical analysis of KCs.RESULTS: UC-MSCs transfused into rats were recruited by the injured liver and alleviated liver fibrosis, increasing serum IL-4 and IL-10 levels. Interestingly, UC-MSCs promoted mobilization of KCs not only in fibrotic livers, but also in vitro. Co-culture of UC-MSCs with KCs resulted in increased production of IL-4 and IL-10. The addition of IL-4 antibodies into the coculture system resulted in decreased KC mobilization.CONCLUSION: UC-MSCs could increase IL-4 and promote mobilization of KCs both in vitro and in vivo, subsequently alleviating the liver fibrosis induced by DMN.展开更多
AIM: To examine the protective effect of green tea extract (GT) on hepatic fi brosis in vitro and in vivo in dimethylnitrosamine (DMN)-induced rats.METHODS: HSC-T6, a rat hepatic stellate cell line, was used as an in ...AIM: To examine the protective effect of green tea extract (GT) on hepatic fi brosis in vitro and in vivo in dimethylnitrosamine (DMN)-induced rats.METHODS: HSC-T6, a rat hepatic stellate cell line, was used as an in vitro assay system. Cell proliferation, collagen content, and type 1 collagen expression were examined in activated HSC-T6 cells. Collagen was determined by estimating the hydroxyproline content. In rats with DMN-induced hepatic fi brosis, serum aspartate aminotransferase and alanine aminotransferase concentrations, liver hydroxyproline and lipid peroxides were determined. Pathologic changes were examined by hematoxylin & eosin staining.RESULTS: GT administration prevented the development of hepatic fibrosis in the rat model of DMN-induced liver fi brosis. These results were confi rmed both by liver histology and by quantitative measurement of hepatic hydroxyproline content, a marker of liver collagen deposition. Accordingly, inhibition of proliferation, reduced collagen deposition, and type 1 collagen expression were observed in activated HSC-T6 cells following GT treatment. These results imply that GT reduced the proliferation of activated HSC and down regulated the collagen content and expression of collagen type 1, thereby ameliorating hepatic fibrosis.CONCLUSION: This study demonstrates that greentea administration can effectively improve liver fibrosis caused by DMN, and may be used as a therapeutic option and preventive measure against hepatic f ibrosis.展开更多
AIM: To determine the gene expression profile data for the whole liver during development of dimethylni-trosamine (DMN)-induced hepatic fibrosis.METHODS: Marker genes were identified for different types of hepatic cel...AIM: To determine the gene expression profile data for the whole liver during development of dimethylni-trosamine (DMN)-induced hepatic fibrosis.METHODS: Marker genes were identified for different types of hepatic cells, including hepatic stellate cells (HSCs), Kupffer cells (including other inflammatory cells), and hepatocytes, using independent temporal DNA microarray data obtained from isolated hepatic cells. RESULTS: The cell-type analysis of gene expression gave several key results and led to formation of three hypotheses: (1) changes in the expression of HSC-specific marker genes during fibrosis were similar to gene expression data in in vitro cultured HSCs, suggesting a major role of the self-activating characteristics of HSCs in formation of fibrosis; (2) expression of mast cell-specific marker genes reached a peak during liver fibrosis, suggesting a possible role of mast cells in formation of fibrosis; and (3) abnormal expression of hepatocyte-specific marker genes was found across several metabolic pathways during fibrosis, including sulfur-containing amino acid metabolism, fatty acid metabolism, and drug metabolism, suggesting a mechanistic relationship between these abnormalities and symptoms of liver fibrosis. CONCLUSION: Analysis of marker genes for specific hepatic cell types can identify the key aspects of fibro-genesis. Sequential activation of inflammatory cells and the self-supporting properties of HSCs play an important role in development of fibrosis.展开更多
AIM: To study the preventive effects of Qianggan-Rongxian Decoction on liver fibrosis induced by dimethylnitrosamine (DMN) in rats. METHODS: Male Wistar rats were randomly divided into hepatic fibrosis model group, co...AIM: To study the preventive effects of Qianggan-Rongxian Decoction on liver fibrosis induced by dimethylnitrosamine (DMN) in rats. METHODS: Male Wistar rats were randomly divided into hepatic fibrosis model group, control group and 3 treatment groups (12 rats in each group). Except for the normal control group, all the rats received 1% DMN (10 μL/kg body weight, i.p), 3 times a week for 4 wk. The rats in the 3 treatment groups including a high-dose DMN group (10 mL/kg), a medium-dose DMN group (7 mL/kg), and a low-dose DMN group (4 mL/kg) were daily gavaged with Qianggan-Rongxian Decoction, and the rats in the model and normal control groups were given saline vehicle. Enzyme-linked immunosorbent assay (ELISA) was used to determine the changes in serum hyaluronic acid (HA), laminin (LN), and type IV collagen levels. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured using routine laboratory methods. Pathologic changes, particularly fibrosis, were examined by hematoxylin and eosin (HE) and Sirius red staining. Hepatic stellate cells (HSC) were examined by transmission electron microscopy. RESULTS: Compared with the model control group, the serum levels of HA, LN, type Ⅳ collagen, ALT and AST were decreased markedly in the other groups after treatment with Qianggan-Rongxian Decoction, especially in the medium-dose DMN group (P < 0.05).Moreover, the area-density percentage of collagen fibrosis was lower in the Qianggan-Rongxian Decoction treatment groups than in the model group, and a more significant drop was observed in the medium-dose DMN group (P < 0.05). CONCLUSION: Qianggan-Rongxian Decoction can inhibit hepatic fibrosis due to chronic liver injury, delay the development of cirrhosis, and notably ameliorate liver function. It may be used as a safe and effective thera-peutic drug for patients with fibrosis.展开更多
[ Objectives ] This study was conducted to evaluate the action of the aqueous extract of a medicinal plant Plumbago zeylanica L. in the reversal of dime- thylnitrosamine （DMN） -induced hepatic fibrosis in rats, and ...[ Objectives ] This study was conducted to evaluate the action of the aqueous extract of a medicinal plant Plumbago zeylanica L. in the reversal of dime- thylnitrosamine （DMN） -induced hepatic fibrosis in rats, and to provide a scientific basis for the utilization of P. zeylanica in treatment of hepatic fibrosis. [Meth- ods ] Hepatic fibrosis in rats was induced by intraperitoneal injection of DMN. The rats were then given the aqueous extract of P. zeylanica at high, medium or low concentration by garage for five weeks. Serum level of alanine aminotransferase （ALT） was determined by lactate dehydrogenase （LDH）-release assay, and serum level of aspartate transaminase （AST） was measured by UV-malate dehydrogenase （MDH） assay. Serum levels of total bilirubin （TBIL）, direct bilirubin （DBIL） and indirect bilirubin （1BIL） were measured by vanadate oxidation assay. Four indices of hepatic fibrosis （hyaluronic acid, laminin, procollagen type III and colla- gen type IV） were determined by radioimmunoassay （RIA） assay. Morphological damage of liver tissue was observed by hematoxylin-eosin staining （H＆E stai- ning）. Immunohistochemical staining was performed to determine the location and area of deposited collagen type I, collagen type III and ct-smeoth muscle actin （a-SMA） in liver tissue. [ Results] Compared with the negative control （rats with diseased fiver and untreated with P. zeylanica aqueous extract）, the serum lev- els of ALT, AST, TBIL, DBIL and IBIL were significantly decreased by P. zeylanica aqueous extract; the levels of the four serum indices of hepatic fibrosis were also obviously reduced. H＆E staining showed that hepatic fibrosis in rats was obviously inhibited or even reversed by P. zeylanica aqueous extract. Immunohisto- chemical staining proved that the aqueous extract of P. zeylanica significantly reduced the area and coloration of collagen type I, collagen type III and ct-SMA in rat liver. [ Conclusions] The aqueous extract of P. zeylanica has a definite effect in reversal of DMN-indueed hepatic fibrosis in rat by promoting the recovery of liver function, reversal of histopathological changes and reducing fibrotic collagen.展开更多
The present study was designed to evaluate the hepatoprotective and antioxidant potentials of silibinin(SBN)against N-nitrosodimethylamine(DMN)-induced toxic insults in the rat liver.The liver damage was induced in Wi...The present study was designed to evaluate the hepatoprotective and antioxidant potentials of silibinin(SBN)against N-nitrosodimethylamine(DMN)-induced toxic insults in the rat liver.The liver damage was induced in Wistar albino rats by repeated administration of DMN(10 mg·kg-1 b.w.,i.p.)on 3 consecutive days per week for 3 weeks.SBN(100 mg·kg-1 b.w.,p.o.)was given daily to the DMN treated rats for two weeks.The marker enzymes of liver toxicity and second-line enzymic and non-enzymic antioxidants were evaluated in serum and liver tissues before and after SBN treatment.Histopathology of the liver was evaluated by H & E staining.The DMN treatment produced a progressive increase in all the serum marker enzymes(AST,ALT,ALP,LDH,and γ-GT),peaking on Day 21.This treatment produced highly significant decreases in all the second-line antioxidant parameters(GSH,GST,GR,GPx,and vitamins C and E).The SBN treatment significantly reversed the DMN-induced damages,towards normalcy.Histopathological studies confirmed the development of liver toxicity in DMN-treated rats,which was reversed by SBN treatment in corroboration with the aforementioned biochemical results,indicating the hepatoprotective and antioxidant properties of SBN.In conclusion,the DMN-induced degenerative changes in the liver were alleviated by SBN treatment and this protective ability may be attributed to its antioxidant,free radical scavenging,and membrane stabilizing properties.展开更多
文摘AIM： To investigate if iron deposition and fat accumulation in the liver play a pathogenetic role in dimethylnitrosamine （DMN）-induced liver fibrosis in rat.METHODS： Thirty rats were treated with DMN at does consecutive days of 10 μL/kg daily, i.p., for 3 consecutive day each week for 4 wk. Rats （n = 30） were sacrificed on the first day （model group A） and 21st d （model group B） after cessation of DMN injection. The control group （n = 10） received an equivalent amount of saline. Liver tissues were stained with hematoxylin ＆ eosin （HE） and Masson and Prussian blue assay and oberserved under electron microscopy. Serum alanine aminotransferase （ALT）and liver tissue hydroxyproline （Hyp） content were tested.RESULTS： The liver fibrosis did not automaticallyreverse, which was similar to previous reports, the perilobular deposition of iron accompanied with collagen showed marked characteristics at both the first and 21st d after cessation of DMN injection. However, fat accumulation in hepatocytes occurred only at the 21^st d after cessation of DMN injection.CONCLUSION： Iron deposition and fat accumulation may play important roles in pathological changes in DMN-induced rat liver fibrosis. The detailed mechanisms of these characteristics need further research.
基金Supported by the Oriental Medicine R and D Project, Ministry of Health and Welfare, Republic of Korea, No. B050018
文摘AIM： To evaluate the therapeutic effect of Chunggan extract （CGX）, a modified traditional Chinese hepatotherapeutic herbal, on the dimethylnitrosamine （DMN）-induced chronic liver injury model in rats. METHODS： Liver injuries were induced in Wistar rats by injection of DMN （ip, 10 mg/mL per kg） for 3 consecutive days per week for 4 wk. The rats were administered with CGX Coo, 100 or 200 mg/kg per day） or distilled water as a control daily for 4 wk starting from the 15^th d of the DMN treatment. Biochemical parameters （serum albumin, bilirubin, ALP, AST and ALT）, lipid peroxides, hydroxyproline, as well as histological changes in liver tissues were analyzed. In addition, gene expression of TNF-α, TGF-β, TIMP-1, TIMP-2, PDGF-β, and MMP-2, all of which are known to be associated with liver fibrosis, were analyzed using real-time PCR. RESULTS： CGX administration restored the spleen weight to normal alter having been increased by DMN treatment. Biochemical analysis of the serum demonstrated that CGX significantly decreased the serum level of ALP （P 〈 0.05）, ALT （P 〈 0.02）, and AST （P 〈 0.02） that had been elevated by DMN treatment. CGX administration moderately lowered lipid peroxide production and markedly lowered hydroxyproline generation caused by DMN treatment in accordance with histopathological examination. DMN treatment induced a highly upregulated expression of TNF-α, TGF-β,TIMP-1, TIMP-2, PDGF-β, and MMP-2. Of these, the gene expression encoding PDGF-β and MMP-2 was still further enhanced 2 wk after secession of the 4-wk DMN treatment, and was remarkably ameliorated by CGX administration. CONCLUSION： CGX exhibits hepatotherapeutic properties against chronic hepatocellular destruction and consequential liver fibrosis.
文摘Objective:To observe the expression level of TNF-α mRNA in rats with hepatic fibrosis induced by dimethylnitrosamine (DMN) and to explore its relationship with collagen metabolism and its diagnostic value for hepatic fibrosis.Methods: Twenty-five male Wistar rats were randomly divided into normal control group (n=10) and model group (n=15). Model rats were induced by DMN for 4 weeks and at final stage were executed. TNF-α mRNA were detected by RT-PCR and the inflammatory necrosis and collagen deposition in hepatic tissue were observed by HE stain and Sirius red stain. The liver functions were determined by automatic biochemical analytic device. The serum marks of liver fibrosis, such as HA, LN and Ⅳ-C were measured with ELISA and RIA. Results: In this study, the rat model of liver fibrosis induced by DMN was successfully constructed. RT-PCR reveals that TNF-α mRNA expression in control group is lower than that of model group. The liver functions of model group were impaired compared with those of the control group (P<0.01). Semi-quantitive analysis revealed that TNF-α/β-actin of normal rats was 0.39±0.12, while 0.93±0.05 of model rats. The concentration of HA (434.44±98.81 vs 252.9±26.59 ng/ml, P<0.01), LN (70.67±6.32 vs 37.90±5.97 ng/ml, P<0.01) and Ⅳ-C (79.39±10.52 vs 21.40±4.17 ng/ml, P<0.01) were significantly increased in the model group as well. Changes of the indexes were similar to the pathological damage of the liver. Conclusion: The results suggested that activation of TNF-α in liver tissues may be the common pathogenic mechanism of liver fibrosis. TNF-α may be a useful index for the diagnosis of hepatic fibrosis which worthies further investigation.
文摘Salvia miltiorrhiza （Sm） is a traditional herbal medicine with multiple effects on various diseases. Its water-soluble parts have been used to produce injectional powder. In this study, liver fibrosis rats were induced by intraperitoneal injection of dimethylnitrosarnine for 3 consecutive days per week for 4 weeks. After 2 weeks, rats in the positive drug group were subcutaneously injected with 8×10^5 IU/kg IFNα2b, while the Sm treatment groups were intraperitoneally injected with 50, 100 and 200 mg/kg solution of Sm injectional powder, respectively, for 6 days per week for 4 weeks. The results showed that either IFNα2b or the Sm injectional powder significantly increased the body weight and liver to spleen ratio, and three doses of the powder brought down the spleen index. Serum analysis showed that both IFNα2b and the Sm powder reduced levels of alanine transaminase and total bilirubin, while only 100 and 200 mg/kg of the Sm powder ameliorated aspartate transaminase and albumin levels. In the collagen examination, reduced hyaluronic acid and procollagen type III levels, less fibrous hyperplasia and collagen deposits, and improved hepatocyte states were clearly observed in rats treated with either IFNα2b or Sm injectional powder. In addition, the mechanism of action of the Sm powder was also studied. Immunohistochemical staining showed that IFNα2b and Sm injectional powder significantly down-regulated the expression of transforming growth factor-β1 （TGF-β1） and platelet derived growth factor （PDGF）. In conclusion, Sm injectional powder has protective effects on dimethylnitrosamine-initiated liver fibrosis in rats, and the mechanism may include the down-regulation of TGF-β1 and PDGF.
基金Supported by the Shanghai Natural Science Foundation(N0.10ZR1430700)Three-Year Plan of Action of Traditional Chinese Medicine in Shanghai(No.ZY3-RCPY-1-1011)
文摘Objective：To study the effect of total flavonoids of Astmgali Radix（TFA）on liver cirrhosis induced with dimethylnitrosamine（DMN）in rats,and the effect on peroxisome proliferator-activated receptorγ（PPARγ）,uncoupling protein 2（UCP2）and farnesoid X receptor（FXR）.Methods：Fifty-three Sprague-Dawley rats were randomly divided into a control group（10 rats）and a DMN group（43 rats）.Rats in the DMN group were given DMN for 4 weeks and divided randomly into a model group（14 rats）,a low-dosage TFA group（14 rats）and a high-dosage TFA group（15 rats）in the 3rd week.Rats were given TFA for 4 weeks at the dosage of 15 and 30 mg/kg in the low-and high-TFA groups,respectively.At the end of the experiment blood and liver samples were collected.Serum liver function and liver tissue hydroxyproline content were determined.hematoxylin-eosin（HE）,Sirus red and immunohistochemical stainings of collagenⅠ,smooth muscle actin（α-SMA）was conducted in paraffinembedded liver tissue slices.Real time polymerase chain reaction（PCR）was adopted to determine PPARγ,UCP2 and FXR m RNA levels.Western blot was adopted to determine protein levels of collagenⅠ,α-SMA,PPARγ,UCP2 and FXR.Results：Compared with the model group,TFA increased the ratio of liver/body weight（low-TFA group P〈0.05,high-TFA group P〈0.01）,improved liver biochemical indices（P〈0.01 for ALT,AST,GGT in both groups,P〈0.05 for albumin and TBil in the high-TFA group）and reduced liver tissue hydroxproline content（P〈0.01 in both groups）in treatment groups significantly.HE staining showed that TFA alleviated liver pathological changes markedly and Sirus red staining showed that TFA reduced collagen deposition,alleviated formation and extent of liver pseudolobule.CollagenⅠandα-SMA immunohistochemical staining showed that staining area and extent markedly decreased in TFA groups compared with the model group.TFA could increase PPARγ,it regulated target UCP2,and FXR levels significantly compared with the model group（in the low-TFA group all P〈0.05,in the high group all P〈0.01）.Conclusions：TFA could improve liver function,alleviate liver pathological changes,and reduce collagen deposition and formation of liver pseudolobule in rats with liver cirrhosis.The antifibrotic effect of TFA was through regulating PPARγsignal pathway and the interaction with FXR.
文摘AIM: To evaluate the antifibrotic effect of different doses of recombinant human Gamma-Interferon (IFN-gamma) in two rat models of hepatic fibrosis, and to observe its effect on moderate chronic hepatitis B virus fibrosis. METHODS: Hepatic fibrosis was successfully induced in 150 and 196 rats by subcutaneous injection of carbon tetrachloride (CCl4) and intraperitoneal injection of dimethylnitrosamine (DMN), respectively. Each of the two model groups was divided into: (1) fibrotic model group; (2) colchicine treatment group (0.1 mg/kg/day, gastrogavage for 8 weeks); (3) high-dose IFN-gamma group (15 MU/kg per day, i.m. for 8 weeks); (4) medium-dose IFN-gamma group (5 MU/kg daily, i.m. for 8 weeks); and (5) Y low-dose IFN-gamma group (1.67 MU/kg daily, i.m. for 8 weeks). Another group of 10 rats without any treatment was used as normal controls. At the end of the experiment, semi-quantitative histopathological scores of inflammation and fibrosis, liver alpha smooth muscle actin (alpha-SMA) expression level, liver hydroxyl proline content and serum hyaluronic acid levels were compared. And 47 medium chronic hepatitis B viral fibrosis patients were studied. They were given IFN-gamma treatment, 100 MU/day i.m. for the first three months and 100 MU qod i.m. for the next six months. Semi-quantitative pathological scores of inflammation and fibrosis and serum hepatic fibrosis indices were compared within the 9 months. RESULTS: In animal experiment, the pathological fibrosis scores and liver hydroxyl proline content were found to be significantly lower in rats treated with different doses of IFN-gamma as compared with rats in fibrotic model group induced by either CCl4 or DMN, in a dose-dependent manner. For CCl4-induced model, pathological fibrosis scores in high, medium and low doses IFN-gamma groups were 5.10 +/- 2.88, 7.70 +/- 3.53 and 8.00 +/- 3.30, respectively, but the score was 14.60 +/- 7.82 in fibrotic model group. Hydroxyl proline contents were 2.83 +/- 1.18, 3.59 +/- 1.22 and 4.80 +/- 1.62, in the three IFN-gamma groups, and 10.01 +/- 3.23 in fibrotic model group. The difference was statistically significant (P【0.01). Similar results were found in DMN-induced model. Pathological fibrosis scores were 6.30 +/- 0.48, 8.10 +/- 2.72 and 8.30 +/- 2.58, in high, medium and low doses IFN-gamma groups, and 12.60 +/- 3.57 in fibrotic model group. Hydroxyl proline contents were 2.72 +/- 0.58, 3.14 +/- 0.71 and 3.62 +/- 1.02, in the three IFN-gamma groups, and 12.79 +/- 1.54 in fibrotic model group. The difference was statistically significant (P【0.01).Serum hepatic fibrosis indices decreased significantly in the 47 patients after IFN-gamma treatment (HA: 433.38 +/- 373.00 vs 281.57 +/- 220.48; LN: 161.22 +/- 41.02 vs 146 +/- 35 +/- 44. 67; PC III: 192.59 +/- 89.95 vs 156.98 +/- 49.22; C-I: 156.30 +/- 44.01 vs 139.14 +/- 34.47) and the differences between the four indices were significant (P 【0.05). Thirty-three patients received two liver biopsies, one before and one after IFN-gamma treatment. In thirty of 33 patients IFN-gamma had better effects according to semi-quantitative pathological scores (8.40 +/- 5.83 vs 5.30 +/- 4.05, P【0.05). CONCLUSION: All the three doses of IFN-gamma are effective in treating rat liver fibrosis induced by either CCl4 or DMN, the higher the dose, the better the effect. And IFN-gamma is effective for patients with moderate chronic hepatitis B viral fibrosis.
基金Supported by National Natural Science Foundation of China,No.81072913
文摘AIM: To evaluate the efficacy of umbilical cord-derived mesenchymal stem cells(UC-MSCs) transplantation in the treatment of liver fibrosis.METHODS: Cultured human UC-MSCs were isolated and transfused into rats with liver fibrosis induced by dimethylnitrosamine(DMN). The effects of UC-MSCs transfusion on liver fibrosis were then evaluated by histopathology; serum interleukin(IL)-4 and IL-10 levels were also measured. Furthermore, Kupffer cells(KCs) in fibrotic livers were isolated and cultured to analyze their phenotype. Moreover, UC-MSCs were cocultured with KCs in vitro to assess the effects of UCMSCs on KCs' phenotype, and IL-4 and IL-10 levels were measured in cell culture supernatants. Finally, UCMSCs and KCs were cultured in the presence of IL-4 antibodies to block the effects of this cytokine, followed by phenotypical analysis of KCs.RESULTS: UC-MSCs transfused into rats were recruited by the injured liver and alleviated liver fibrosis, increasing serum IL-4 and IL-10 levels. Interestingly, UC-MSCs promoted mobilization of KCs not only in fibrotic livers, but also in vitro. Co-culture of UC-MSCs with KCs resulted in increased production of IL-4 and IL-10. The addition of IL-4 antibodies into the coculture system resulted in decreased KC mobilization.CONCLUSION: UC-MSCs could increase IL-4 and promote mobilization of KCs both in vitro and in vivo, subsequently alleviating the liver fibrosis induced by DMN.
文摘AIM: To examine the protective effect of green tea extract (GT) on hepatic fi brosis in vitro and in vivo in dimethylnitrosamine (DMN)-induced rats.METHODS: HSC-T6, a rat hepatic stellate cell line, was used as an in vitro assay system. Cell proliferation, collagen content, and type 1 collagen expression were examined in activated HSC-T6 cells. Collagen was determined by estimating the hydroxyproline content. In rats with DMN-induced hepatic fi brosis, serum aspartate aminotransferase and alanine aminotransferase concentrations, liver hydroxyproline and lipid peroxides were determined. Pathologic changes were examined by hematoxylin & eosin staining.RESULTS: GT administration prevented the development of hepatic fibrosis in the rat model of DMN-induced liver fi brosis. These results were confi rmed both by liver histology and by quantitative measurement of hepatic hydroxyproline content, a marker of liver collagen deposition. Accordingly, inhibition of proliferation, reduced collagen deposition, and type 1 collagen expression were observed in activated HSC-T6 cells following GT treatment. These results imply that GT reduced the proliferation of activated HSC and down regulated the collagen content and expression of collagen type 1, thereby ameliorating hepatic fibrosis.CONCLUSION: This study demonstrates that greentea administration can effectively improve liver fibrosis caused by DMN, and may be used as a therapeutic option and preventive measure against hepatic f ibrosis.
文摘AIM: To determine the gene expression profile data for the whole liver during development of dimethylni-trosamine (DMN)-induced hepatic fibrosis.METHODS: Marker genes were identified for different types of hepatic cells, including hepatic stellate cells (HSCs), Kupffer cells (including other inflammatory cells), and hepatocytes, using independent temporal DNA microarray data obtained from isolated hepatic cells. RESULTS: The cell-type analysis of gene expression gave several key results and led to formation of three hypotheses: (1) changes in the expression of HSC-specific marker genes during fibrosis were similar to gene expression data in in vitro cultured HSCs, suggesting a major role of the self-activating characteristics of HSCs in formation of fibrosis; (2) expression of mast cell-specific marker genes reached a peak during liver fibrosis, suggesting a possible role of mast cells in formation of fibrosis; and (3) abnormal expression of hepatocyte-specific marker genes was found across several metabolic pathways during fibrosis, including sulfur-containing amino acid metabolism, fatty acid metabolism, and drug metabolism, suggesting a mechanistic relationship between these abnormalities and symptoms of liver fibrosis. CONCLUSION: Analysis of marker genes for specific hepatic cell types can identify the key aspects of fibro-genesis. Sequential activation of inflammatory cells and the self-supporting properties of HSCs play an important role in development of fibrosis.
基金the Research Program of Department of Scienceand Technology of Hebei Province, No. 200721047
文摘AIM: To study the preventive effects of Qianggan-Rongxian Decoction on liver fibrosis induced by dimethylnitrosamine (DMN) in rats. METHODS: Male Wistar rats were randomly divided into hepatic fibrosis model group, control group and 3 treatment groups (12 rats in each group). Except for the normal control group, all the rats received 1% DMN (10 μL/kg body weight, i.p), 3 times a week for 4 wk. The rats in the 3 treatment groups including a high-dose DMN group (10 mL/kg), a medium-dose DMN group (7 mL/kg), and a low-dose DMN group (4 mL/kg) were daily gavaged with Qianggan-Rongxian Decoction, and the rats in the model and normal control groups were given saline vehicle. Enzyme-linked immunosorbent assay (ELISA) was used to determine the changes in serum hyaluronic acid (HA), laminin (LN), and type IV collagen levels. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured using routine laboratory methods. Pathologic changes, particularly fibrosis, were examined by hematoxylin and eosin (HE) and Sirius red staining. Hepatic stellate cells (HSC) were examined by transmission electron microscopy. RESULTS: Compared with the model control group, the serum levels of HA, LN, type Ⅳ collagen, ALT and AST were decreased markedly in the other groups after treatment with Qianggan-Rongxian Decoction, especially in the medium-dose DMN group (P < 0.05).Moreover, the area-density percentage of collagen fibrosis was lower in the Qianggan-Rongxian Decoction treatment groups than in the model group, and a more significant drop was observed in the medium-dose DMN group (P < 0.05). CONCLUSION: Qianggan-Rongxian Decoction can inhibit hepatic fibrosis due to chronic liver injury, delay the development of cirrhosis, and notably ameliorate liver function. It may be used as a safe and effective thera-peutic drug for patients with fibrosis.
基金Supported by National Natural Science Foundation of China(81403189,81460628,81660705,81560690)Scientific Research Project for Higher Education of Guangxi Zhuang Autonomous Region Education Department(YB2014182)
文摘[ Objectives ] This study was conducted to evaluate the action of the aqueous extract of a medicinal plant Plumbago zeylanica L. in the reversal of dime- thylnitrosamine （DMN） -induced hepatic fibrosis in rats, and to provide a scientific basis for the utilization of P. zeylanica in treatment of hepatic fibrosis. [Meth- ods ] Hepatic fibrosis in rats was induced by intraperitoneal injection of DMN. The rats were then given the aqueous extract of P. zeylanica at high, medium or low concentration by garage for five weeks. Serum level of alanine aminotransferase （ALT） was determined by lactate dehydrogenase （LDH）-release assay, and serum level of aspartate transaminase （AST） was measured by UV-malate dehydrogenase （MDH） assay. Serum levels of total bilirubin （TBIL）, direct bilirubin （DBIL） and indirect bilirubin （1BIL） were measured by vanadate oxidation assay. Four indices of hepatic fibrosis （hyaluronic acid, laminin, procollagen type III and colla- gen type IV） were determined by radioimmunoassay （RIA） assay. Morphological damage of liver tissue was observed by hematoxylin-eosin staining （H＆E stai- ning）. Immunohistochemical staining was performed to determine the location and area of deposited collagen type I, collagen type III and ct-smeoth muscle actin （a-SMA） in liver tissue. [ Results] Compared with the negative control （rats with diseased fiver and untreated with P. zeylanica aqueous extract）, the serum lev- els of ALT, AST, TBIL, DBIL and IBIL were significantly decreased by P. zeylanica aqueous extract; the levels of the four serum indices of hepatic fibrosis were also obviously reduced. H＆E staining showed that hepatic fibrosis in rats was obviously inhibited or even reversed by P. zeylanica aqueous extract. Immunohisto- chemical staining proved that the aqueous extract of P. zeylanica significantly reduced the area and coloration of collagen type I, collagen type III and ct-SMA in rat liver. [ Conclusions] The aqueous extract of P. zeylanica has a definite effect in reversal of DMN-indueed hepatic fibrosis in rat by promoting the recovery of liver function, reversal of histopathological changes and reducing fibrotic collagen.
基金supported by the DST-INSPIRE(Department of Science and Technology,Government of India Innovation in Science Pursuit for Inspired Research)fellowship(Award No:DST/INSPIRE Fellowship/2010/dated 16.03.2010)
文摘The present study was designed to evaluate the hepatoprotective and antioxidant potentials of silibinin(SBN)against N-nitrosodimethylamine(DMN)-induced toxic insults in the rat liver.The liver damage was induced in Wistar albino rats by repeated administration of DMN(10 mg·kg-1 b.w.,i.p.)on 3 consecutive days per week for 3 weeks.SBN(100 mg·kg-1 b.w.,p.o.)was given daily to the DMN treated rats for two weeks.The marker enzymes of liver toxicity and second-line enzymic and non-enzymic antioxidants were evaluated in serum and liver tissues before and after SBN treatment.Histopathology of the liver was evaluated by H & E staining.The DMN treatment produced a progressive increase in all the serum marker enzymes(AST,ALT,ALP,LDH,and γ-GT),peaking on Day 21.This treatment produced highly significant decreases in all the second-line antioxidant parameters(GSH,GST,GR,GPx,and vitamins C and E).The SBN treatment significantly reversed the DMN-induced damages,towards normalcy.Histopathological studies confirmed the development of liver toxicity in DMN-treated rats,which was reversed by SBN treatment in corroboration with the aforementioned biochemical results,indicating the hepatoprotective and antioxidant properties of SBN.In conclusion,the DMN-induced degenerative changes in the liver were alleviated by SBN treatment and this protective ability may be attributed to its antioxidant,free radical scavenging,and membrane stabilizing properties.
