Effect of bone marrow mesenchymal stem cells on the Smad expression of hepatic fibrosis rats

Objective:To investigate the impact of bone marrow mesenchymal stem cells on Smad expression of hepatic fibrosis rats.Methods:A total of 48 adult female SD rats were randomly divided into three groups,normal control group(n=10),observation group(n=19)with liver fibrosis model rats injected with BMSCs cells:model group(n=19),with liver fibrosis model rats injected with physiological saline.Serum index,TGF-β1 and Smad expression were detected.Results:TypeⅢprocollagen,Ⅳcollagen,hyaluronic acid,laminin levels of observation group were significantly lower than those of model group(P0.05).The content and expression of TGF-β1in serum and liver tissue of observation group were significantly lower than those of model group(P0.05).Compared with normal control group,the Smad3,Smad4 mRNA and protein expression of model group were significantly increased,the Smad7 mRNA and protein expression were significantly reduced(P0.05).Compared with model group.Smad3,Smad4 mRNA and protein expression of observation group were significantly reduced,and Smad7 mRNA expression were significantly increased(P0.05).Conclusions:BMSCs can regulate Smad expression to some extent,and reduce the degree of liver fibrosis.     

Treatment of pig serum-induced rat liver fibrosis with Boschniakia rossica, oxymatrine and interferon-α

AIM: To investigate the effect of Boschniakia rossica (BR), oxymatrine (OM) and interferon-alpha (IFN-α) 1b on the therapy of rat liver fibrosis and its mechanism. METHODS: By establishing a rat model of pig serum-induced liver fibrosis, liver/weight index and serum alanine transaminase (ALT) were observed to investigate the therapeutic effect of BR,OM and IFN-α. Radioimmunoassay was utilized to measure procollagen type Ⅲ (PCⅢ) and collagen type Ⅳ (CIV). RT-PCR was used to assay the expression of liver transforming growth factor- beta 1 (TGF-β1) mRNA. Immunohistochemistry of alpha-smooth muscle actin (α-SMA) and pathologic changes of liver tissues were also under investigation. RESULTS: Serum PCⅢ and CIV in BR, OM and IFN-α groups were significantly declined compared with those in model group, and their RT-PCR revealed that TGF-β1 mRNA expression was also reduced more than that in model group. Immunohistochemistry demonstrated that α-SMA also declined more than that in model group. Serum ALT in IFN-α, control and model groups was within normal level. Serum ALT in BR group had no significant difference from those of IFN-α, control and model groups. Serum ALT in OM group was significantly higher than those in BR, IFN-α, model, and control groups. CONCLUSION: BR, OM and IFN-α can prevent pig serum-induced liver rat fibrosis by inhibiting the activation of hepatic stellate cells and synthesizing collagen. OM has hepatotoxicity to rat liver fibrosis induced by pig serum.     

Therapeutic effects and molecular mechanisms of anti-fibrosis herbs and selenium on rats with hepatic fibrosis

AIM:To study the therapeutic effects of anti-fibrosis herbsand selenium on hepatic fibrosis induced by carbontetrachloride (CCl_4) in rats and the underlining molecularmechanisms.METHODS:Fifty-three Wistar rats were randomly dividedinto:normal control group,model control group,colchicinegroup,anti-fibrosis herbs group (AF group) and anti-fibrosisherbs plus selenium group (AS group).The last four groupswere administered with CCl_4 at the beginning of experimentto induce hepatic fibrosis.Then colchicine,anti-fibrosis herbsand selenium were used to treat them.The normal controlgroup and the model control group were given normal salineat the same time.At the end of the 6~(th) week,rats in eachgroup were sacrificed.Blood and tissue specimens weretaken.Serum indicators (ALT,AST,HA,LN) were determinedand histopathological changes were graded.Lymphocyte CD_4and CD_8 were examined by flow cytometry.Expression ofTGF-β_1 and NF-κB was detected by immunohistochemistryand expression of TGF-β_1 mRNA was detected by semi-quantified RT-PCR.RESULTS:Histological grading showed much a smallerdegree of hepatic fibrogenesis in AS group and AF groupthan that in colchicine group and model control group.Theserum content of ALT,AST,HA and LN in AF group and ASgroup were significantly lower than that in colchicine group(ALT:65.8±26.5,67.3±18.4 and 96.2±20.9 in AF,AS andcolchicine groups respectively;AST:150.8±34.0,154.6±27.3and 215.8±24.6 respectively;HA:228±83,216±58 and416±135 respectively;LN:85.9±15.0,80.6±18.6 and106.3±14.2 respectively) (P<0.05).The level of CD_4 andCD_4/CD_8 ratio in AF group and AS group was significantly higherthat those in cochicine group (CD_4:50.8±3.8,52.6±3.4 and40.2±2.1 in AF,AS and colchicine groups respectively;CD_4/CD_8 ratio:1.45,1.46 and 1.26,respectively (P<0.05).The expression level of NF-κB and TGF-β_1 in the liver tissuesof AF and AS treatment groups was markedly decreasedcompared with that in cochicine group,and TGF-β_1 mRNAwas also markedly decreased (1.07±0.31 and 0.98±0.14 vs2.34±0.43,P<0.05).CONCLUSION:Anti-fibrosis herbs and selenium havebeneficial effects on hepatic fibrosis in rats by enhancingimmunity and inhibiting NF-κB and TGF-β_1 expressions.     

Treatment of pig serum-induced rat liver fibrosis with Boschniakia rossica, oxymatrine and interferon-α

AIM: To investigate the effect of Boschniakia rossica (BR), oxymatrine (OM) and interferon-alpha (IFN-α) lb on the therapy of rat liver fibrosis and its mechanism. METHODS: By establishing a rat model of pig serum-induced liver fibrosis, liver/weight index and serum alanine transaminase (ALT) were observed to investigate the therapeutic effect of BR, OM and IFN-α Radioimmunoassay was utilized to measure procollagen type Ⅲ (PCⅢ) and collagen type Ⅳ (CⅣ). RT-PCR was used to assay the expression of liver transforming growth factor- beta 1 (TGF-β1) mRNA. Immunohistochemistry of alpha-smooth muscle actin (α-SMA) and pathologic changes of liver tissues were also under investigation. RESULTS: Serum PCⅢ and CⅣ in BR, OM and IFN-α groups were significantly declined compared with those in model group, and their RT-PCR revealed that TGF-β1 mRNA expression was also reduced more than that in model group. Immunohistochemistry demonstrated that α-SMA also declined more than that in model group. Serum ALT in IFN-α, control and model groups was within normal level. Serum ALT in BR group had no significant difference from those of IFN-α, control and model groups. Serum ALT in ON group was significantly higher than those in BR, IFN-α,model, and control groups. CONCLUSION: BR, OM and IFN-α can prevent pig serum-induced liver rat fibrosis by inhibiting the activation of hepatic stellate cells and synthesizing collagen. OM has hepatotoxicity to rat liver fibrosis induced by pig serum.     

Effect of matrine on transforming growth factor β1 and hepatocyte growth factor in rat liver fibrosis model

Objective:To observe the preventive and control effect of matrine on transforming growth factor(TCF- β1) and hepatocyte.growth factor(HCF) of liver fibrosis tissue in rals.Methods:A total of48 SD rats were randomly divided into A,B,C,D groups with 12 in each,group A as the normal control group and groups B.C,D as liver fibrosis models using composite modulus method with carbon tetrachloride(CCL_4).Group B was the model group,group C adopted γ— interferon lavage therapy in the second day of modeling,and group D adopted matrine lavage treatment,at 4 and8 weeks after treatment.Six rats were executed for detection of TGF- β1 and HGF,liver tissue histology and comparison fibrosis degree changes of rat liver tissue between groups.Results:Croups B,C,D showed a more significantly increased TCF- β1 at each time point compared with group A(P0.05);Group B showed a more significantly increased TGF- β1 than groups C and D at weeks 4 and 8(P0.05);group D showed a lowest level of TGF-β1,followed by groups C and B.HGF of group B decreased more significantly than A group at weeks 4 and 8(P0.05);HGF of groups C and D was significantly elevated at 4 and 8 weeks than groups A and B(P0.05),in which the group D showed the highest level of HGF.According to tissue histologic observation,rat liver tissue structure of group A was clear and normal,tissue structure of group B was destroyed with obvious fibrous tissue hyperplasia and fatty change of hepatic cells;groups C and D showed a slighter liver tissue damage,cell necrosis and connective tissue hyperplasia in collect abbacy than group B with a trend of obvious improvement.Conclusions:Matrine can reduce TGF- β1expression and enhance the activity of HGF,so as to realize the inhibition effect on liver fibrosis in rats.     

ACEI attenuates the progression of CCl_4-induced rat hepatic fibrogenesis by inhibiting TGF-β1, PDGF-BB, NF-κB and MMP-2,9

AIM: Angiotensin II has pro-fibrotic function in the liver. Blockade of the renin-angiotensin-aldosterone-system (RMS) attenuates hepatic fibrosis. The aim of the present study was to determine the mechanism of angiotensin-converting enzyme inhibitor (ACEI) on the progression of rat hepatic fibrosis. METHODS: Forty male Wistar rats were divided into three groups. Model group (Mo): The rats were injected subcutaneously with 40% of CCl4 0.25 mL/100 g. Perindopril group (Pe): The rats were injected subcutaneously with 40% of CCl4. Perindopril, equivalent to 2 mg/(kg·d), was administrated. Control group (Nc): the rats were treated with olive oil only. After 4 and 6 wk, the rats were killed. The liver sections were stained with Masson. The protein expressions of AT1R, TGF-β1 and PDGF-BB were examined by Western blot. Nuclear factor κB (NF-κB) DNA binding activity was examined by EMSA (Electrophoretic gel mobility shift assay). Matrix metalloproteinase-2,9 (MMP-2,9) activity was assessed by zymography. Serum laminin (LN) and hyaluronic acid (HA) were measured using radioimmunoassays. RESULTS: Using Western blot, we clearly provided direct evidence for the expression of AT1R in liver. The expression was up-regulated when fibrogenesis occurred. Perindopril treatment significantly reduced mean fibrosis score, protein levels of AT1R, TGF-β1 and PDGF-BB, serum levels of HA and LN, and the activity of MMP-2,9. NPκB DNA binding activity markedly increased in model group, perindopril treatment considerably reduced NPκB DNA binding activity. CONCLUSION: Perindopril attenuates CCl4-induced hepatic fibrogenesis of rat by inhibiting TGF-β1, PDGF-BB, NF-κB and MMP-2,9.     

Effect of NLRC5 on activation and reversion of hepatic stellate cells by regulating the nuclear factor-κB signaling pathway

BACKGROUND The formation of liver fibrosis is mainly caused by the activation of hepatic stellate cells(HSCs)and the imbalance of extracellular matrix(ECM)production and degradation.The treatment of liver fibrosis mainly includes removing the cause,inhibiting the activation of HSCs,and inhibiting inflammation.NOD-like receptor(NLR)family,caspase activation and recruitment domain(CARD)domain containing 5/NOD27/CLR16.1(NLRC5)is a highly conserved member of the NLR family and is involved in inflammation and immune responses by regulating various signaling pathways such as nuclear factor-κB(NF-κB)signaling.It has been found that NLRC5 plays an important role in liver fibrosis,but its specific effect and possible mechanism remain to be fully elucidated.AIM To investigate the role of NLRC5 in the activation and reversion of HSCs induced with transforming growth factor-β(TGF-β)and MDI,and to explore its relationship with liver fibrosis.METHODS A total of 24 male C57BL/6 mice were randomly divided into three groups,including normal,fibrosis,and recovery groups.Twenty-four hours after a liver fibrosis and spontaneous reversion model was established,the mice were sacrificed and pathological examination of liver tissue was performed to observe the degree of liver fibrosis in each group.LX-2 cells were cultured in vitro and treated with TGF-β1 and MDI.Real-time quantitative PCR(qPCR)and Western blot were used to analyze the expression levels of NLRC5,α-smooth muscle actin(α-SMA),and collagen type I alpha1(Col1a1)in each group.The activity of NF-κB in each group of cells transfected with NLRC5-siRNA was detected.RESULTS Compared with the normal mice,the expression level of NLRC5 increased significantly(P<0.01)in the fibrosis group,but decreased significantly in the recovery group(P<0.01).In in vitro experiments,the content of NLRC5 was enhanced after TGF-β1 stimulation and decreased to a lower level when treated with MDI(P<0.01).The expression ofα-SMA and Col1a1 proteins and mRNAs in TGF-β1-mediated cells was suppressed by transfection with NLRC5-siRNA(P<0.01).Western blot analysis showed that the expression of NF-κB p65 protein and phosphorylated IκBα(p-IκBα)was increased in the liver of mice in the fibrosis group but decreased in the recovery group(P<0.01),and the protein level of nuclear p65 and p-IκBαwas significantly increased after treatment with NLRC5-siRNA(P<0.01).CONCLUSION NLRC5 may play a key role in the development and reversal of hepatic fibrosis through the NF-κB signaling pathway,and it is expected to be one of the clinical therapeutic targets.     

Therapeutic effect of interleukin-10 on CCI_4-induced hepatic fibrosis in rats

AIM: To study the therapeutic effect of exogenous inter-leukin-10 on CCI4-induced hepatic fibrosis in rats and its possible mechanisms. METHODS: Fourty-seven SD rats were randomly divided into control group (group N) and CCI4-induced hepatic fibrosis model group (group C). After CCI4 was given for 9 wk, the model group was divided into three groups. Rats in group M were put to death immediately, rats in group T were treated with IL-10 for another three wk and then put to death, rats in group R recovered after three weeks and were then killed. The degree of hepatic fibrosis was measured by HE staining and histological activity index (HAI). Histological activity index (HAI), change of collagen typesⅠandⅢwere measured by Picrosirius staining. The expression of TNF-α, MMP-2 and TIMP-1 in liver tissue was measured by S-P immunohis-tochemistry. RESULTS: CCI4- induced experimental rat hepatic fibrosis model was established successfully. The degree of hepatic fibrosis was markedly lower in group T than in groups M and R, and there was no difference between the two groups. The expression of collagen typesⅠandⅢwas significantly suppressed in group T and was slightly suppressed in groups M and R. The positive levels of TNF-α, MMP-2 and TIMP-1 in group M increased significantly compared to those in group N (P<0.01). The positive signals decreased significantly in groups T and R (P<0.01), but positive score was significantly lower in group T than in group R (P< 0.01). CONCLUSION: Exogenous IL-10 can reverse CCI4-in-duced hepatic fibrosis in rats. IL-10 may exert its reversible effects on hepatic fibrosis by blocking CCI4-induced inflammation, inhibiting expression of MMP-2 and TIMP-1 and promoting resolution of collagen typesⅠandⅢ.     

The effects of gossypol on expressions of transforming growth factor-β1, fibronectin and 11β-hydroxysteroid dehydrogenase in nephridial tissue in rats with diabetes mellitus

Objective To observe the effects of gossypol on expressions of transforming growth factor-β1 (TGF-β1), fibronectin (FN) and 11β-hydroxysteroid dehydrogenase (11β-HSD) in nephridial tissue in rats with diabetes mellitus. Methods Thirty male Sprague-Dawley rats were randomly divided into three groups: normal control, type 2 diabetes and gossypol treatment group . After high-fat feeding for 4 weeks, the later two groups were injected with low dosage strepozotocin (30 mg/kg) intraperitoneally to induce type 2 diabetic rat model. The rats in gossypol treated group were given gossypol at the dosage of 15 mg/kg once per day for 4 weeks by gavage. And since the 5th week, the times of gavages had been changed into once per week at the same dosage and lasted to the 12th week . The levels of blood glucose, total cholesterol (TC), low density lipoprotein-cholesterol (LDL-c) were measured. Additionally, the morphological changes of the kidney were studied by light microscopy and transmission electron microscopy respectively. The mRNA expressions of TGF-β1, FN, 11β-HSD and 11β-HSD2 in nephridial tissue were assayed by semi-quantity RT-PCR. The protein expressions of TGF-β1 and FN in nephridial tissue were determined by immunohistochemistry. Results The blood levels of glucose, TC and LDL- c were increased significantly in type 2 diabetic group compared with normal control group(P<0. 01). The volume of glomerulus and the deposition of PAS positive substance in the glomerular interstitium were increased under light microscopy, and the glomerular basal membrane was thicker in type 2 diabetic group than those in normal control group under transmission electron microscopy. The mRNA and protein expressions of TGF-β1 and FN were increased(P<0. 01), and the mRNA expression of 11β-HSD2 was decreased(P<0. 05), while the mRNA expression of 11β-HSD1 was unchanged in type 2 diabetic group compared with normal control group. After the treatment of gossypol, the level of the blood glucose was significantly decreased(P< 0. 01), and the levels of TC, LDL-c showed a trend of decrease but had no statistical differences compared with type 2 diabetic group. The morphology of nephridial tissue was ameliorated in gossypol treatment group. The mRNA and protein expressions of TGF-β1 and FN were decreased(0. 16± 0. 02,0. 22±0. 05 ; 0. 24±0. 06,0. 33±0. 07, P< 0. 05), while the mRNA expressions of 11β-HSD1 and 11β-HSD2 were unchanged compared with type 2 diabetic group. Conclusions Gossypol can relieve the pathologic changes of nephridial tissue, inhibit the expressions of TGF-β1 and FN through decreasing blood glucose of rats with diabetes mellitus.     

The effects of gossypol on expressions of transforming growth factor-β1, fibronectin and 11β-hydroxysteroid dehydrogenase in nephridial tissue in rats with diabetes mellitus

Objective To observe the effects of gossypol on expressions of transforming growth factor-β1 (TGF-β1), fibronectin (FN) and 11β-hydroxysteroid dehydrogenase (11β-HSD) in nephridial tissue in rats with diabetes mellitus. Methods Thirty male Sprague-Dawley rats were randomly divided into three groups: normal control, type 2 diabetes and gossypol treatment group . After high-fat feeding for 4 weeks, the later two groups were injected with low dosage strepozotocin (30 mg/kg) intraperitoneally to induce type 2 diabetic rat model. The rats in gossypol treated group were given gossypol at the dosage of 15 mg/kg once per day for 4 weeks by gavage. And since the 5th week, the times of gavages had been changed into once per week at the same dosage and lasted to the 12th week . The levels of blood glucose, total cholesterol (TC), low density lipoprotein-cholesterol (LDL-c) were measured. Additionally, the morphological changes of the kidney were studied by light microscopy and transmission electron microscopy respectively. The mRNA expressions of TGF-β1, FN, 11β-HSD and 11β-HSD2 in nephridial tissue were assayed by semi-quantity RT-PCR. The protein expressions of TGF-β1 and FN in nephridial tissue were determined by immunohistochemistry. Results The blood levels of glucose, TC and LDL- c were increased significantly in type 2 diabetic group compared with normal control group(P<0. 01). The volume of glomerulus and the deposition of PAS positive substance in the glomerular interstitium were increased under light microscopy, and the glomerular basal membrane was thicker in type 2 diabetic group than those in normal control group under transmission electron microscopy. The mRNA and protein expressions of TGF-β1 and FN were increased(P<0. 01), and the mRNA expression of 11β-HSD2 was decreased(P<0. 05), while the mRNA expression of 11β-HSD1 was unchanged in type 2 diabetic group compared with normal control group. After the treatment of gossypol, the level of the blood glucose was significantly decreased(P< 0. 01), and the levels of TC, LDL-c showed a trend of decrease but had no statistical differences compared with type 2 diabetic group. The morphology of nephridial tissue was ameliorated in gossypol treatment group. The mRNA and protein expressions of TGF-β1 and FN were decreased(0. 16± 0. 02,0. 22±0. 05 ; 0. 24±0. 06,0. 33±0. 07, P< 0. 05), while the mRNA expressions of 11β-HSD1 and 11β-HSD2 were unchanged compared with type 2 diabetic group. Conclusions Gossypol can relieve the pathologic changes of nephridial tissue, inhibit the expressions of TGF-β1 and FN through decreasing blood glucose of rats with diabetes mellitus.     

The effects of gossypol on expressions of transforming growth factor-β1, fibronectin and 11β-hydroxysteroid dehydrogenase in nephridial tissue in rats with diabetes mellitus

Objective To observe the effects of gossypol on expressions of transforming growth factor-β1 (TGF-β1), fibronectin (FN) and 11β-hydroxysteroid dehydrogenase (11β-HSD) in nephridial tissue in rats with diabetes mellitus. Methods Thirty male Sprague-Dawley rats were randomly divided into three groups: normal control, type 2 diabetes and gossypol treatment group . After high-fat feeding for 4 weeks, the later two groups were injected with low dosage strepozotocin (30 mg/kg) intraperitoneally to induce type 2 diabetic rat model. The rats in gossypol treated group were given gossypol at the dosage of 15 mg/kg once per day for 4 weeks by gavage. And since the 5th week, the times of gavages had been changed into once per week at the same dosage and lasted to the 12th week . The levels of blood glucose, total cholesterol (TC), low density lipoprotein-cholesterol (LDL-c) were measured. Additionally, the morphological changes of the kidney were studied by light microscopy and transmission electron microscopy respectively. The mRNA expressions of TGF-β1, FN, 11β-HSD and 11β-HSD2 in nephridial tissue were assayed by semi-quantity RT-PCR. The protein expressions of TGF-β1 and FN in nephridial tissue were determined by immunohistochemistry. Results The blood levels of glucose, TC and LDL- c were increased significantly in type 2 diabetic group compared with normal control group(P<0. 01). The volume of glomerulus and the deposition of PAS positive substance in the glomerular interstitium were increased under light microscopy, and the glomerular basal membrane was thicker in type 2 diabetic group than those in normal control group under transmission electron microscopy. The mRNA and protein expressions of TGF-β1 and FN were increased(P<0. 01), and the mRNA expression of 11β-HSD2 was decreased(P<0. 05), while the mRNA expression of 11β-HSD1 was unchanged in type 2 diabetic group compared with normal control group. After the treatment of gossypol, the level of the blood glucose was significantly decreased(P< 0. 01), and the levels of TC, LDL-c showed a trend of decrease but had no statistical differences compared with type 2 diabetic group. The morphology of nephridial tissue was ameliorated in gossypol treatment group. The mRNA and protein expressions of TGF-β1 and FN were decreased(0. 16± 0. 02,0. 22±0. 05 ; 0. 24±0. 06,0. 33±0. 07, P< 0. 05), while the mRNA expressions of 11β-HSD1 and 11β-HSD2 were unchanged compared with type 2 diabetic group. Conclusions Gossypol can relieve the pathologic changes of nephridial tissue, inhibit the expressions of TGF-β1 and FN through decreasing blood glucose of rats with diabetes mellitus.     

The effects of gossypol on expressions of transforming growth factor-β1, fibronectin and 11β-hydroxysteroid dehydrogenase in nephridial tissue in rats with diabetes mellitus

Objective To observe the effects of gossypol on expressions of transforming growth factor-β1 (TGF-β1), fibronectin (FN) and 11β-hydroxysteroid dehydrogenase (11β-HSD) in nephridial tissue in rats with diabetes mellitus. Methods Thirty male Sprague-Dawley rats were randomly divided into three groups: normal control, type 2 diabetes and gossypol treatment group . After high-fat feeding for 4 weeks, the later two groups were injected with low dosage strepozotocin (30 mg/kg) intraperitoneally to induce type 2 diabetic rat model. The rats in gossypol treated group were given gossypol at the dosage of 15 mg/kg once per day for 4 weeks by gavage. And since the 5th week, the times of gavages had been changed into once per week at the same dosage and lasted to the 12th week . The levels of blood glucose, total cholesterol (TC), low density lipoprotein-cholesterol (LDL-c) were measured. Additionally, the morphological changes of the kidney were studied by light microscopy and transmission electron microscopy respectively. The mRNA expressions of TGF-β1, FN, 11β-HSD and 11β-HSD2 in nephridial tissue were assayed by semi-quantity RT-PCR. The protein expressions of TGF-β1 and FN in nephridial tissue were determined by immunohistochemistry. Results The blood levels of glucose, TC and LDL- c were increased significantly in type 2 diabetic group compared with normal control group(P<0. 01). The volume of glomerulus and the deposition of PAS positive substance in the glomerular interstitium were increased under light microscopy, and the glomerular basal membrane was thicker in type 2 diabetic group than those in normal control group under transmission electron microscopy. The mRNA and protein expressions of TGF-β1 and FN were increased(P<0. 01), and the mRNA expression of 11β-HSD2 was decreased(P<0. 05), while the mRNA expression of 11β-HSD1 was unchanged in type 2 diabetic group compared with normal control group. After the treatment of gossypol, the level of the blood glucose was significantly decreased(P< 0. 01), and the levels of TC, LDL-c showed a trend of decrease but had no statistical differences compared with type 2 diabetic group. The morphology of nephridial tissue was ameliorated in gossypol treatment group. The mRNA and protein expressions of TGF-β1 and FN were decreased(0. 16± 0. 02,0. 22±0. 05 ; 0. 24±0. 06,0. 33±0. 07, P< 0. 05), while the mRNA expressions of 11β-HSD1 and 11β-HSD2 were unchanged compared with type 2 diabetic group. Conclusions Gossypol can relieve the pathologic changes of nephridial tissue, inhibit the expressions of TGF-β1 and FN through decreasing blood glucose of rats with diabetes mellitus.     

The effects of gossypol on expressions of transforming growth factor-β1, fibronectin and 11β-hydroxysteroid dehydrogenase in nephridial tissue in rats with diabetes mellitus

Objective To observe the effects of gossypol on expressions of transforming growth factor-β1 (TGF-β1), fibronectin (FN) and 11β-hydroxysteroid dehydrogenase (11β-HSD) in nephridial tissue in rats with diabetes mellitus. Methods Thirty male Sprague-Dawley rats were randomly divided into three groups: normal control, type 2 diabetes and gossypol treatment group . After high-fat feeding for 4 weeks, the later two groups were injected with low dosage strepozotocin (30 mg/kg) intraperitoneally to induce type 2 diabetic rat model. The rats in gossypol treated group were given gossypol at the dosage of 15 mg/kg once per day for 4 weeks by gavage. And since the 5th week, the times of gavages had been changed into once per week at the same dosage and lasted to the 12th week . The levels of blood glucose, total cholesterol (TC), low density lipoprotein-cholesterol (LDL-c) were measured. Additionally, the morphological changes of the kidney were studied by light microscopy and transmission electron microscopy respectively. The mRNA expressions of TGF-β1, FN, 11β-HSD and 11β-HSD2 in nephridial tissue were assayed by semi-quantity RT-PCR. The protein expressions of TGF-β1 and FN in nephridial tissue were determined by immunohistochemistry. Results The blood levels of glucose, TC and LDL- c were increased significantly in type 2 diabetic group compared with normal control group(P<0. 01). The volume of glomerulus and the deposition of PAS positive substance in the glomerular interstitium were increased under light microscopy, and the glomerular basal membrane was thicker in type 2 diabetic group than those in normal control group under transmission electron microscopy. The mRNA and protein expressions of TGF-β1 and FN were increased(P<0. 01), and the mRNA expression of 11β-HSD2 was decreased(P<0. 05), while the mRNA expression of 11β-HSD1 was unchanged in type 2 diabetic group compared with normal control group. After the treatment of gossypol, the level of the blood glucose was significantly decreased(P< 0. 01), and the levels of TC, LDL-c showed a trend of decrease but had no statistical differences compared with type 2 diabetic group. The morphology of nephridial tissue was ameliorated in gossypol treatment group. The mRNA and protein expressions of TGF-β1 and FN were decreased(0. 16± 0. 02,0. 22±0. 05 ; 0. 24±0. 06,0. 33±0. 07, P< 0. 05), while the mRNA expressions of 11β-HSD1 and 11β-HSD2 were unchanged compared with type 2 diabetic group. Conclusions Gossypol can relieve the pathologic changes of nephridial tissue, inhibit the expressions of TGF-β1 and FN through decreasing blood glucose of rats with diabetes mellitus.     

The effects of gossypol on expressions of transforming growth factor-β1, fibronectin and 11β-hydroxysteroid dehydrogenase in nephridial tissue in rats with diabetes mellitus

Objective To observe the effects of gossypol on expressions of transforming growth factor-β1 (TGF-β1), fibronectin (FN) and 11β-hydroxysteroid dehydrogenase (11β-HSD) in nephridial tissue in rats with diabetes mellitus. Methods Thirty male Sprague-Dawley rats were randomly divided into three groups: normal control, type 2 diabetes and gossypol treatment group . After high-fat feeding for 4 weeks, the later two groups were injected with low dosage strepozotocin (30 mg/kg) intraperitoneally to induce type 2 diabetic rat model. The rats in gossypol treated group were given gossypol at the dosage of 15 mg/kg once per day for 4 weeks by gavage. And since the 5th week, the times of gavages had been changed into once per week at the same dosage and lasted to the 12th week . The levels of blood glucose, total cholesterol (TC), low density lipoprotein-cholesterol (LDL-c) were measured. Additionally, the morphological changes of the kidney were studied by light microscopy and transmission electron microscopy respectively. The mRNA expressions of TGF-β1, FN, 11β-HSD and 11β-HSD2 in nephridial tissue were assayed by semi-quantity RT-PCR. The protein expressions of TGF-β1 and FN in nephridial tissue were determined by immunohistochemistry. Results The blood levels of glucose, TC and LDL- c were increased significantly in type 2 diabetic group compared with normal control group(P<0. 01). The volume of glomerulus and the deposition of PAS positive substance in the glomerular interstitium were increased under light microscopy, and the glomerular basal membrane was thicker in type 2 diabetic group than those in normal control group under transmission electron microscopy. The mRNA and protein expressions of TGF-β1 and FN were increased(P<0. 01), and the mRNA expression of 11β-HSD2 was decreased(P<0. 05), while the mRNA expression of 11β-HSD1 was unchanged in type 2 diabetic group compared with normal control group. After the treatment of gossypol, the level of the blood glucose was significantly decreased(P< 0. 01), and the levels of TC, LDL-c showed a trend of decrease but had no statistical differences compared with type 2 diabetic group. The morphology of nephridial tissue was ameliorated in gossypol treatment group. The mRNA and protein expressions of TGF-β1 and FN were decreased(0. 16± 0. 02,0. 22±0. 05 ; 0. 24±0. 06,0. 33±0. 07, P< 0. 05), while the mRNA expressions of 11β-HSD1 and 11β-HSD2 were unchanged compared with type 2 diabetic group. Conclusions Gossypol can relieve the pathologic changes of nephridial tissue, inhibit the expressions of TGF-β1 and FN through decreasing blood glucose of rats with diabetes mellitus.     

Effects of interferon-alpha on expression of hepatic stellate cell and transforming growth factor-β1 and α-smooth muscle actin in rats with hepatic fibrosis

AIM: To investigate the effect of interferon-α(IFN-α) on preventing or reversing hepatic fibrosis in rat experimental model induced by CCl4.METHODS: One hundred and ten Sprague-Dawley rats were divided into five groups: group A (normal controls,n=18), group B (fibrotic model controls, n=22), group C (IFN-α prevention, n=22) initially treated with intra-muscular injection of IFN-α in saline daily at the doses of 1&#215;105U for 6wk, group D (IFN-α treatment, n=24) treated with intra-muscular injection of IFN-α in saline daily at the doses of 1&#215;105U for 6wk after the first 6wk, group E (0.9% sodium chloride treatment control, n=24) treated with intra-muscular injection of 0.01mL/kg daily for 6wk after the first 6wk. At the end of the experiment, all rats of each group were killed. Samples of the liver obtained by biopsy were subjected to histological, immunohistochemical and electron microscopic studies for the expressions of transforming growth factor-β1(TGF-β1) and α-smooth muscle actin (α-SMA).RESULTS: The expressions of TGF-β1, the number of activated hepatic stellate cells and α-SMA in hepatic tissue of group C were significantly less than those of group B(P&lt;0.01). The degree of fibrosis score in group B was also significantly less than that of group C under light microscope (P&lt;0.01).CONCLUSION: IFN-α can inhibit the production of TGF-β1, decrease HSC activation and stimulate its apoptosis.     

Notch signaling mediated by TGF-β/Smad pathway in concanavalin A-induced liver fibrosis in rats

AIM To explore the exact interaction between Notch and transforming growth factor(TGF)-β signaling in liver fibrosis. METHODS We established a rat model of liver fibrosis induced by concanavalin A. Peripheral blood mononuclear cells(PBMCs) were isolated from the modeled rats, and cultured with γ-secretase inhibitor DAPT and TGF-β inhibitor for 24 h. The m RNA levels of Notch and TGF-β signaling were detected by quantitative real-time polymerase chain reaction. Expression of Notch and TGF-β proteins was analyzed by western blotting.RESULTS Compared to control rats, Notch and TGF-β signaling was activated in PBMCs of model rats. Administration of DAPT and TGF-β inhibitor suppressed Notch and TGF-β signal transducer in PBMCs of model rats. DAPT reduced m RNA and protein expression of TGF-β signaling, such as TGF-β1 and Smad3. TGF-β inhibitor also downregulated Notch1, Hes1 and Hes5, and m RNA and protein expression of the Notch signaling pathway.CONCLUSION Notch and TGF-β signaling play a role in liver fibrosis. TGF-β signaling upregulates Notch signaling, which promotes TGF-β signaling.     

Bone morphogenetic protein-7 represses hepatic stellate cell activation and liver fibrosis via regulation of TGF-β/Smad signaling pathway

BACKGROUND Liver fibrosis is a refractory disease whose persistence can eventually induce cirrhosis or even liver cancer.Early liver fibrosis is reversible by intervention.As a member of the transforming growth factor-beta(TGF-β)superfamily,bone morphogenetic protein 7(BMP7)has anti-liver fibrosis functions.However,little is known about BMP7 expression changes and its potential regulatory mechanism as well as the relationship between BMP7 and TGF-βduring liver fibrosis.In addition,the mechanism underlying the anti-liver fibrosis function of BMP7 needs to be further explored.AIM To investigate changes in the dynamic expression of BMP7 during liver fibrosis,interactions between BMP7 and TGF-β1,and possible mechanisms underlying the anti-liver fibrosis function of BMP7.METHODS Changes in BMP7 expression during liver fibrosis and the interaction between BMP7 and TGF-β1 in mice were observed.Exogenous BMP7 was used to treat mouse primary hepatic stellate cells(HSCs)to observe its effect on activation,migration,and proliferation of HSCs and explore the possible mechanism underlying the anti-liver fibrosis function of BMP7.Mice with liver fibrosis received exogenous BMP7 intervention to observe improvement of liver fibrosis by using Masson’s trichrome staining and detecting the expression of the HSC activation indicator alpha-smooth muscle actin(α-SMA)and the collagen formation associated protein type I collagen(Col I).Changes in the dynamic expression of BMP7 during liver fibrosis in the human body were further observed.RESULTS In the process of liver fibrosis induced by carbon tetrachloride(CCl4)in mice,BMP7 protein expression first increased,followed by a decrease;there was a similar trend in the human body.This process was accompanied by a sustained increase in TGF-β1 protein expression.In vitro experiment results showed that TGF-β1 inhibited BMP7 expression in a time-and dose-dependent manner.In contrast,high doses of exogenous BMP7 inhibited TGF-β1-induced activation,migration,and proliferation of HSCs;this inhibitory effect was associated with upregulation of pSmad1/5/8 and downregulation of phosphorylation of Smad3 and p38 by BMP7.In vivo experiment results showed that exogenous BMP7 improved liver fibrosis in mice.CONCLUSION During liver fibrosis,BMP7 protein expression first increases and then decreases.This changing trend is associated with inhibition of BMP7 expression by sustained upregulation of TGF-β1 in a time-and dose-dependent manner.Exogenous BMP7 could selectively regulate TGF-β/Smad pathway-associated factors to inhibit activation,migration,and proliferation of HSCs and exert antiliver fibrosis functions.Exogenous BMP7 has the potential to be used as an antiliver fibrosis drug.