Transforming growth factor-β (TGF-β), a prototype of multifunctional cytokine, is a key regulator of extracellular matrix (ECM) assembly and remodeling. Specifically, TGF-β isoforms have the ability to induce the e...Transforming growth factor-β (TGF-β), a prototype of multifunctional cytokine, is a key regulator of extracellular matrix (ECM) assembly and remodeling. Specifically, TGF-β isoforms have the ability to induce the expression of ECM proteins in mesenchymal cells, and to stimulate the production of protease inhibitors that prevent enzymatic breakdown of the ECM. Elevated TGF-β expression in affected organs, and subsequent deregulation of TGF-β functions, correlates with the abnormal connective tissue deposition observed during the onset of fibrotic diseases. During the last few years, tremendous progress has been made in the understanding of the molecular aspects of intracellular signaling downstream of the TGF-β receptors. In particular, Smad proteins, TGF-β receptor kinase substrates that translocate into the cell nucleus to act as transcription factors, have been studied extensively. The role of Smad3 in the transcriptional regulation of typeⅠcollagen gene expression and in the development of fibrosis, demonstrated both in vitro and in animal models with a targeted deletion of Smad3, is of critical importance because it may lead to novel therapeutic strategies against these diseases. This review focuses on the mechanisms underlying Smad modulation of fi brillar collagen expression and how it relates to fi brotic processes.展开更多
AIM: To investigate the inhibition effects on the productionof collagen type I, Ⅲ secreted by activated rat hepatic stellatecells (rHSCs) by antisense tissue inhibitors of metalloproteinase1 (TIMP-1) recombinant plas...AIM: To investigate the inhibition effects on the productionof collagen type I, Ⅲ secreted by activated rat hepatic stellatecells (rHSCs) by antisense tissue inhibitors of metalloproteinase1 (TIMP-1) recombinant plasmid through elevating interstitialcollagenase activity.METHODS: rHSCs were extracted from normal rat liverby pronase and collagenase digestion and purified bycentrifugal elutriation, and were cultured on plastic dishesuntil they were activated to a myofibroblastic phenotypeafter 7-10 days. RT-Nest-PCR and gene recombinanttechniques were used to construct the rat antisense TIMP-1 recombinant plasmids which can express in eucaryoticcells. The recombinant plasmid and the pcDNA3 emptyplasmid were transfected in rHSCs by Effectene (QIAGEN)separately. Cells were selected after growing in DMEMcontaining 400 μg/ml G418 for 2-3 weeks. Expression ofexogenous gene was assessed by Northern blot, andexpression oflIMP-1 in rHSCs was determined by Northernblot and Western blot. We tested the interstitial collagenaseactivity with FITC-labled type I collagen as substrate.Ultimately, we quantified the type Ⅰ, Ⅲ collagen byWestern blot.RESULTS: The exogenous antisense TIMP-1 recombinantplasmid could be expressed in rHSCs well, which couldblock the expression of TIMP-1 greatly, the ratio of TIMP-1/GAPDH was 0.67, 2.41, and 2.97 separately at mRNAlevel (P＜0.05); the ratio of TIMP-1/β-actin was 0.31, 0.98and 1.32 separately at protein level (P＜0.05); It mightelevate active and latent interstitial collagenase activity,the collagenase activity was 0.3049, 0.1411 and 0.1196respectively. (P＜0.05), which led to promotion thedegradation of type Ⅰ, Ⅲ collagen, the ratio of collagen I/β-actin was 0.63, 1.78 and 1.92 separately (P＜0.05); andthe ratio of collagen Ⅲ/β-actin was 0.59, 1.81 and 1.98separately (P＜0.05).CONCLUSION: These data shows that the antisense TIMP-1 recombinant plasmid has the inhibitory effects on theproduction of type Ⅰ, Ⅲ collagens secreted by activatedrHSCs in vitro. It could be a novel method to reverse hepaticfibrosis in the future.展开更多
AIM: To study the effect of oxymatrine-baicalin combination (OB) against HBV replication in 2.2.15 cells and a smooth muscle actin (αSMA) expression, typeⅠ, collagen synthesis in HSC-T6 cells. METHODS: The 2.2.15 ce...AIM: To study the effect of oxymatrine-baicalin combination (OB) against HBV replication in 2.2.15 cells and a smooth muscle actin (αSMA) expression, typeⅠ, collagen synthesis in HSC-T6 cells. METHODS: The 2.2.15 cells and HSC-T6 cells were cultured and treated respectively. HBsAg and HBeAg in the culture supernatants were detected by ELISA and HBV DNA levels were determined by fluorescence quantitative PCR. Total RNA was extracted from HSC-T6 cells and reverse transcribed into cDNA. The cDNAs were amplified by PCR and the quantities were expressed in proportion to p actin. The total cellular proteins extracted from HSC-T6 cells were separated by electrophoresis. Resolved proteins were electrophoretically transferred to nitrocellulose membrane. Protein bands were revealed and the quantities were corrected by p actin. RESULTS: In the 2.2.15 cell culture system, the inhibitory rate against secretion of HBsAg and HBeAg in the OB group was significantly stronger than that in the oxymatrine group (HBsAg, P=0.043; HBeAg, P=0.026; respectively); HBV DNA level in the OB group was significantly lower than that in the oxymatrine group (P = 0.041). In HSC-T6 cells the mRNA and protein expression levels of a SMA in the OB group were significantly lower as compared with those in the oxymatrine group (mRNA, P = 0.013; protein, P-0.042; respectively); The mRNA and protein expression levels of typeⅠcollagen in the OB group were significantly lower as compared with those in the oxymatrine group (mRNA, P<0.01; protein, P<0.01; respectively). CONCLUSION: OB combination has a better effect against HBV replication in 2.2.15 cells and is more effective against a SMA expression and typeⅠcollagen synthesis in HSC-T6 cells than oxymatrine in vitro.展开更多
AIM: To explore the effects of endothelin-1(ET-1) on hepatic stellate cells (HSCs) DNA uptake, DNA synthesis,collagen synthesis and secretion, inward whole-cell calcium concentration ([Ca^2+]i) as well as the blocking...AIM: To explore the effects of endothelin-1(ET-1) on hepatic stellate cells (HSCs) DNA uptake, DNA synthesis,collagen synthesis and secretion, inward whole-cell calcium concentration ([Ca^2+]i) as well as the blocking effect of verapamil on ET-l-stimulated release of inward calcium(Ca2+) of HSC in vitro.METHODS: Rat hepatic stellate cells (HSCs) were isolated and cultivated. ^3H-TdR and ^3H-proline incorporation used for testing DNA uptake and synthesis, collagen synthesis and secretion of HSCs cultured in vitro;, Fluorescent calcium indicator Fura-2/AM was used to measure [Ca^2+]i inwardHSCs.RESULTS: ET-1 at the concentration of 5×10^-8 mol/L,caused significant increase both in HSC DNA synthesis (2 247+344 cpm, P<0.05) and DNA uptake (P<0.05) when compared with the control group. ET-1 could also increase collagen synthesis (P<0.05 vs control group) and collagen secretion (P<0.05 vscontrol group). Besides, inward HSC [Ca^2+]i reached a peak concentration (422±98 mol/L, P<0.001)at 2 min and then went down slowly to165+51 mol/L(P<0.01) at 25 min from resting state (39±4 mol/L)after treated with ET-1. Verapamil (5 mol/L) blocked ET-1-activated [Ca^2+li inward HSCs compared with control group(P<0.05). Fura-2/AM loaded HSC was suspended in no Ca^2+ buffer containing 1 mol/L EGTA, 5 rain later, 10^-8 mol/L of ET-1 was added, [Ca^2+]i inward HSCs rose from resting state to peak 399±123 mol/L, then began to come down by the time of 20 min. It could also raise [Ca^2+]i inward HSCs even without Ca^2+ in extracellular fluid, and had a remarkable dose-effect relationship(P<0.05). Meanwhile,verapamil could restrain the action of ET-1(P<0.05).CONCLUSION: Actions of ET-1 on collagen metabolism of HSCs may depend on the transportation of inward wholecell calcium.展开更多
AIM: Matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) are central to the spontaneous resolution of liver fibrosis. The mechanisms involved have been investigated in hepati...AIM: Matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) are central to the spontaneous resolution of liver fibrosis. The mechanisms involved have been investigated in hepatic steilate cells (ISC), but not in hepatocytes. We investigated the effects of fibril- and fixed-collagen on MMP-1 and TIMP-1 production in hepatocytes, using the HLE cell line.METHODS: Fibril type T and Ⅳ collagen were prepared by HCl digestion of type T and Ⅳ collagen, respectively.For fixed-collagen, culture dishes were coated with fibril type Ⅰ or Ⅳ collagen and fixed by ultraviolet. Type Ⅰcollagenase activity was measured using fluorescein isothiocyanate-labeled type Ⅰ collagen. MMP-1 and TIMP-1 in HLE cells were measured by a one-step sandwich enzyme immunoassay.RESULTS: Both fibril type Ⅰand Ⅳ collagen significantly increased type Ⅰ coilagenase activity about two-fold compared with no fibril collagen. The effects of the fibril collagen were not affected by the coating condition. There was no significant difference in the effects on collagenase activity between cells cultured in medium containing fibril type Ⅰ collagen and those cultured in the presence of type Ⅳ collagen. Both types of fibril collagen significantly increased MMP-1 production, and showed more than 10-fold higher levels of MMP-1 than the control. The enhanced MMP-1 production by fibril collagens was unaffected by the coating condition. By contrast, TIMP-1 production was not changed by the addition of fibril type Ⅰ or Ⅳ collagen,and neither was it affected by the coating conditions.Coating with type Ⅰ collagen significantly suppressed MMP-1production by almost one-tenth compared with no coating.By contrast, lIMP-1 production was not affected by either the absence of a collagen coat or by increasing the concentration of the coating collagen.CONCLUSION: These results indicated that, in HLE cells,fibril- and fixed-collagen have opposite effects on MMP-1production without affecting TIMP production. Fibril collagen induced collagenase activity by up-regulation of MMP-1 production without affecting TIMP-1 production.By contrast, fixed collagen reduced MMP-1 production.Our results suggest that hepatocytes might also play an important role in the regulation of the hepatic fibrosis alongside HSC.展开更多
AIM: To study the effects of pentoxifylline (PTX) on thecontent of hepatic TGF-β1, type Ⅰ and type Ⅲ collagen inschistosomiasis japonica mice with liver fibrosis and itsmechanism of anti-fibrosis.METHODS: Forty mic...AIM: To study the effects of pentoxifylline (PTX) on thecontent of hepatic TGF-β1, type Ⅰ and type Ⅲ collagen inschistosomiasis japonica mice with liver fibrosis and itsmechanism of anti-fibrosis.METHODS: Forty mice with schistosomiasis were dividedinto four groups: one group as control without anytreatment, other three were treated with Praziquantel 500mg/(kg.d)for 2 d, high dose PTX 360 mg/(kg.d) for 8 wk,and low dose PTX 180 mg/(kg.d) for 8 wk respectively.Immunohistochemical technique and multimedia colorpathographic analysis system were applied to observe thecontent change of hepatic TGF-β1, type Ⅰ and type Ⅲcollagen in schistosomiasis japonica mice with liver fibrosisbefore and after PTX treatment.RESULTS: Effects of PTX on the content change of hepaticTGF-β1, type Ⅰ and type Ⅲ collagen in schistosomiasis japonicamice with liver fibrosis were related to the dosage of PTX,high dose PTX treated group could significantly reduce thecontent of TGF-β1 (0.709±0.111), type Ⅰ (0.644±0.108) andtype Ⅲ (0.654±0.152) collagen compared with those ofcontrol group (0.883±0.140, 0.771±0.156, 0.822±0.129)with statistical significance (P＜0.05). Low dose PTX couldalso reduce the hepatic content of TGF-β1 (0.752±0.152),type Ⅰ (0.733±0.117) and type Ⅲ (0.788±0.147) collagen,but without statistical significance (P＞0.05). Both high doseand low dose PTX groups have significant differences onthe content of TGF-β1, type Ⅰ and type Ⅲ collagen (P＜0.05,P＜0.05, P＜ 0.01,respectively).CONCLUSION: High dose of PTX treatment could reducethe content of hepatic TGF-β1, type Ⅰ and type Ⅲ collagensignificantly in schistosomiasis japonica mice with liverfibrosis, and thus plays its role of antifibrosis.展开更多
AIM: To investigate the role of nuclear factor-κB (NF-κB)inhibitor caffeic acid phenethy1 ester (CAPE) in the proliferation, collagen synthesis and apoptosis of hepatic stellate cells (HSCs) of rats.
METHODS: The HS...AIM: To investigate the role of nuclear factor-κB (NF-κB)inhibitor caffeic acid phenethy1 ester (CAPE) in the proliferation, collagen synthesis and apoptosis of hepatic stellate cells (HSCs) of rats.
METHODS: The HSCs from rats were isolated and cultured in Dulbecco's Modified Eagle's Medium (DMEM) and treated with CAPE. The proliferation and collagen synthesis of HSCs were determined by 3H-TdR and 3H-proline incorporation respectively, and the expression of type Ⅰ, Ⅲ procollagen genes was further explored byin situ hybridization. Apoptosis cell indices (AIs) were examined using terminal deoxynucleotidyl transferase- mediated DIG-dUTP nick end labeling (TUNEL).
RESULTS: Tn activated HSC in culture, CAPE significantly inhibited 3H-TdR and 3H-proline incorporation by HSCs at concentrations of 5 μmol/L and 10 μmol/L respectively. CAPE also reduced the type I procollagen gene expression (P＜0.05)at higher concentration. Apoptosis of HSC was induced by CAPE and the AIs were time-and dose-dependently increased from 2.82+0.73 % to 7.66±1.25 % at 12 h (P＜0.01) and from 3.15±0.88 % to 10.6L±2.88 % at 24 h (P＜0.01).
CONCLUSION: CAPE inhibits proliferation and collagen synthesis of HSC at lower concentration and induces HSC apoptosis at higher concentration.展开更多
基金Programme National de Recherche Dermatologie 2006, Institut Nationale de la Santé Et de la Recherche Médicale, Groupe Franais de Recherche sur la Sclérodermie, and Associa-tion des Slérodermiques de France
文摘Transforming growth factor-β (TGF-β), a prototype of multifunctional cytokine, is a key regulator of extracellular matrix (ECM) assembly and remodeling. Specifically, TGF-β isoforms have the ability to induce the expression of ECM proteins in mesenchymal cells, and to stimulate the production of protease inhibitors that prevent enzymatic breakdown of the ECM. Elevated TGF-β expression in affected organs, and subsequent deregulation of TGF-β functions, correlates with the abnormal connective tissue deposition observed during the onset of fibrotic diseases. During the last few years, tremendous progress has been made in the understanding of the molecular aspects of intracellular signaling downstream of the TGF-β receptors. In particular, Smad proteins, TGF-β receptor kinase substrates that translocate into the cell nucleus to act as transcription factors, have been studied extensively. The role of Smad3 in the transcriptional regulation of typeⅠcollagen gene expression and in the development of fibrosis, demonstrated both in vitro and in animal models with a targeted deletion of Smad3, is of critical importance because it may lead to novel therapeutic strategies against these diseases. This review focuses on the mechanisms underlying Smad modulation of fi brillar collagen expression and how it relates to fi brotic processes.
基金the National Natural Scientific Foundation of China, No.39970339
文摘AIM: To investigate the inhibition effects on the productionof collagen type I, Ⅲ secreted by activated rat hepatic stellatecells (rHSCs) by antisense tissue inhibitors of metalloproteinase1 (TIMP-1) recombinant plasmid through elevating interstitialcollagenase activity.METHODS: rHSCs were extracted from normal rat liverby pronase and collagenase digestion and purified bycentrifugal elutriation, and were cultured on plastic dishesuntil they were activated to a myofibroblastic phenotypeafter 7-10 days. RT-Nest-PCR and gene recombinanttechniques were used to construct the rat antisense TIMP-1 recombinant plasmids which can express in eucaryoticcells. The recombinant plasmid and the pcDNA3 emptyplasmid were transfected in rHSCs by Effectene (QIAGEN)separately. Cells were selected after growing in DMEMcontaining 400 μg/ml G418 for 2-3 weeks. Expression ofexogenous gene was assessed by Northern blot, andexpression oflIMP-1 in rHSCs was determined by Northernblot and Western blot. We tested the interstitial collagenaseactivity with FITC-labled type I collagen as substrate.Ultimately, we quantified the type Ⅰ, Ⅲ collagen byWestern blot.RESULTS: The exogenous antisense TIMP-1 recombinantplasmid could be expressed in rHSCs well, which couldblock the expression of TIMP-1 greatly, the ratio of TIMP-1/GAPDH was 0.67, 2.41, and 2.97 separately at mRNAlevel (P＜0.05); the ratio of TIMP-1/β-actin was 0.31, 0.98and 1.32 separately at protein level (P＜0.05); It mightelevate active and latent interstitial collagenase activity,the collagenase activity was 0.3049, 0.1411 and 0.1196respectively. (P＜0.05), which led to promotion thedegradation of type Ⅰ, Ⅲ collagen, the ratio of collagen I/β-actin was 0.63, 1.78 and 1.92 separately (P＜0.05); andthe ratio of collagen Ⅲ/β-actin was 0.59, 1.81 and 1.98separately (P＜0.05).CONCLUSION: These data shows that the antisense TIMP-1 recombinant plasmid has the inhibitory effects on theproduction of type Ⅰ, Ⅲ collagens secreted by activatedrHSCs in vitro. It could be a novel method to reverse hepaticfibrosis in the future.
文摘AIM: To study the effect of oxymatrine-baicalin combination (OB) against HBV replication in 2.2.15 cells and a smooth muscle actin (αSMA) expression, typeⅠ, collagen synthesis in HSC-T6 cells. METHODS: The 2.2.15 cells and HSC-T6 cells were cultured and treated respectively. HBsAg and HBeAg in the culture supernatants were detected by ELISA and HBV DNA levels were determined by fluorescence quantitative PCR. Total RNA was extracted from HSC-T6 cells and reverse transcribed into cDNA. The cDNAs were amplified by PCR and the quantities were expressed in proportion to p actin. The total cellular proteins extracted from HSC-T6 cells were separated by electrophoresis. Resolved proteins were electrophoretically transferred to nitrocellulose membrane. Protein bands were revealed and the quantities were corrected by p actin. RESULTS: In the 2.2.15 cell culture system, the inhibitory rate against secretion of HBsAg and HBeAg in the OB group was significantly stronger than that in the oxymatrine group (HBsAg, P=0.043; HBeAg, P=0.026; respectively); HBV DNA level in the OB group was significantly lower than that in the oxymatrine group (P = 0.041). In HSC-T6 cells the mRNA and protein expression levels of a SMA in the OB group were significantly lower as compared with those in the oxymatrine group (mRNA, P = 0.013; protein, P-0.042; respectively); The mRNA and protein expression levels of typeⅠcollagen in the OB group were significantly lower as compared with those in the oxymatrine group (mRNA, P<0.01; protein, P<0.01; respectively). CONCLUSION: OB combination has a better effect against HBV replication in 2.2.15 cells and is more effective against a SMA expression and typeⅠcollagen synthesis in HSC-T6 cells than oxymatrine in vitro.
基金Supported by the Grant for Nature and Science from Shanghai,No.03ZR 14097
文摘AIM: To explore the effects of endothelin-1(ET-1) on hepatic stellate cells (HSCs) DNA uptake, DNA synthesis,collagen synthesis and secretion, inward whole-cell calcium concentration ([Ca^2+]i) as well as the blocking effect of verapamil on ET-l-stimulated release of inward calcium(Ca2+) of HSC in vitro.METHODS: Rat hepatic stellate cells (HSCs) were isolated and cultivated. ^3H-TdR and ^3H-proline incorporation used for testing DNA uptake and synthesis, collagen synthesis and secretion of HSCs cultured in vitro;, Fluorescent calcium indicator Fura-2/AM was used to measure [Ca^2+]i inwardHSCs.RESULTS: ET-1 at the concentration of 5×10^-8 mol/L,caused significant increase both in HSC DNA synthesis (2 247+344 cpm, P<0.05) and DNA uptake (P<0.05) when compared with the control group. ET-1 could also increase collagen synthesis (P<0.05 vs control group) and collagen secretion (P<0.05 vscontrol group). Besides, inward HSC [Ca^2+]i reached a peak concentration (422±98 mol/L, P<0.001)at 2 min and then went down slowly to165+51 mol/L(P<0.01) at 25 min from resting state (39±4 mol/L)after treated with ET-1. Verapamil (5 mol/L) blocked ET-1-activated [Ca^2+li inward HSCs compared with control group(P<0.05). Fura-2/AM loaded HSC was suspended in no Ca^2+ buffer containing 1 mol/L EGTA, 5 rain later, 10^-8 mol/L of ET-1 was added, [Ca^2+]i inward HSCs rose from resting state to peak 399±123 mol/L, then began to come down by the time of 20 min. It could also raise [Ca^2+]i inward HSCs even without Ca^2+ in extracellular fluid, and had a remarkable dose-effect relationship(P<0.05). Meanwhile,verapamil could restrain the action of ET-1(P<0.05).CONCLUSION: Actions of ET-1 on collagen metabolism of HSCs may depend on the transportation of inward wholecell calcium.
文摘AIM: Matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) are central to the spontaneous resolution of liver fibrosis. The mechanisms involved have been investigated in hepatic steilate cells (ISC), but not in hepatocytes. We investigated the effects of fibril- and fixed-collagen on MMP-1 and TIMP-1 production in hepatocytes, using the HLE cell line.METHODS: Fibril type T and Ⅳ collagen were prepared by HCl digestion of type T and Ⅳ collagen, respectively.For fixed-collagen, culture dishes were coated with fibril type Ⅰ or Ⅳ collagen and fixed by ultraviolet. Type Ⅰcollagenase activity was measured using fluorescein isothiocyanate-labeled type Ⅰ collagen. MMP-1 and TIMP-1 in HLE cells were measured by a one-step sandwich enzyme immunoassay.RESULTS: Both fibril type Ⅰand Ⅳ collagen significantly increased type Ⅰ coilagenase activity about two-fold compared with no fibril collagen. The effects of the fibril collagen were not affected by the coating condition. There was no significant difference in the effects on collagenase activity between cells cultured in medium containing fibril type Ⅰ collagen and those cultured in the presence of type Ⅳ collagen. Both types of fibril collagen significantly increased MMP-1 production, and showed more than 10-fold higher levels of MMP-1 than the control. The enhanced MMP-1 production by fibril collagens was unaffected by the coating condition. By contrast, TIMP-1 production was not changed by the addition of fibril type Ⅰ or Ⅳ collagen,and neither was it affected by the coating conditions.Coating with type Ⅰ collagen significantly suppressed MMP-1production by almost one-tenth compared with no coating.By contrast, lIMP-1 production was not affected by either the absence of a collagen coat or by increasing the concentration of the coating collagen.CONCLUSION: These results indicated that, in HLE cells,fibril- and fixed-collagen have opposite effects on MMP-1production without affecting TIMP production. Fibril collagen induced collagenase activity by up-regulation of MMP-1 production without affecting TIMP-1 production.By contrast, fixed collagen reduced MMP-1 production.Our results suggest that hepatocytes might also play an important role in the regulation of the hepatic fibrosis alongside HSC.
基金the Science Research Foundation of Schistosomiasis of Hubei Province,No.2000
文摘AIM: To study the effects of pentoxifylline (PTX) on thecontent of hepatic TGF-β1, type Ⅰ and type Ⅲ collagen inschistosomiasis japonica mice with liver fibrosis and itsmechanism of anti-fibrosis.METHODS: Forty mice with schistosomiasis were dividedinto four groups: one group as control without anytreatment, other three were treated with Praziquantel 500mg/(kg.d)for 2 d, high dose PTX 360 mg/(kg.d) for 8 wk,and low dose PTX 180 mg/(kg.d) for 8 wk respectively.Immunohistochemical technique and multimedia colorpathographic analysis system were applied to observe thecontent change of hepatic TGF-β1, type Ⅰ and type Ⅲcollagen in schistosomiasis japonica mice with liver fibrosisbefore and after PTX treatment.RESULTS: Effects of PTX on the content change of hepaticTGF-β1, type Ⅰ and type Ⅲ collagen in schistosomiasis japonicamice with liver fibrosis were related to the dosage of PTX,high dose PTX treated group could significantly reduce thecontent of TGF-β1 (0.709±0.111), type Ⅰ (0.644±0.108) andtype Ⅲ (0.654±0.152) collagen compared with those ofcontrol group (0.883±0.140, 0.771±0.156, 0.822±0.129)with statistical significance (P＜0.05). Low dose PTX couldalso reduce the hepatic content of TGF-β1 (0.752±0.152),type Ⅰ (0.733±0.117) and type Ⅲ (0.788±0.147) collagen,but without statistical significance (P＞0.05). Both high doseand low dose PTX groups have significant differences onthe content of TGF-β1, type Ⅰ and type Ⅲ collagen (P＜0.05,P＜0.05, P＜ 0.01,respectively).CONCLUSION: High dose of PTX treatment could reducethe content of hepatic TGF-β1, type Ⅰ and type Ⅲ collagensignificantly in schistosomiasis japonica mice with liverfibrosis, and thus plays its role of antifibrosis.
基金Natural Science Foundation of Yunnan Province for Younth,No.1999C0034Q,No.2000C0031Q
文摘AIM: To investigate the role of nuclear factor-κB (NF-κB)inhibitor caffeic acid phenethy1 ester (CAPE) in the proliferation, collagen synthesis and apoptosis of hepatic stellate cells (HSCs) of rats.
METHODS: The HSCs from rats were isolated and cultured in Dulbecco's Modified Eagle's Medium (DMEM) and treated with CAPE. The proliferation and collagen synthesis of HSCs were determined by 3H-TdR and 3H-proline incorporation respectively, and the expression of type Ⅰ, Ⅲ procollagen genes was further explored byin situ hybridization. Apoptosis cell indices (AIs) were examined using terminal deoxynucleotidyl transferase- mediated DIG-dUTP nick end labeling (TUNEL).
RESULTS: Tn activated HSC in culture, CAPE significantly inhibited 3H-TdR and 3H-proline incorporation by HSCs at concentrations of 5 μmol/L and 10 μmol/L respectively. CAPE also reduced the type I procollagen gene expression (P＜0.05)at higher concentration. Apoptosis of HSC was induced by CAPE and the AIs were time-and dose-dependently increased from 2.82+0.73 % to 7.66±1.25 % at 12 h (P＜0.01) and from 3.15±0.88 % to 10.6L±2.88 % at 24 h (P＜0.01).
CONCLUSION: CAPE inhibits proliferation and collagen synthesis of HSC at lower concentration and induces HSC apoptosis at higher concentration.
