OBJECTIVE:To study the effect of Lichong decoction(LD) on expression of matrix metalloproteinase-2(MMP-2) and metalloproteinase-2(TIMP-2) in a rat model of uterine leiomyoma(UL).METHODS:UL was induced in rats using ex...OBJECTIVE:To study the effect of Lichong decoction(LD) on expression of matrix metalloproteinase-2(MMP-2) and metalloproteinase-2(TIMP-2) in a rat model of uterine leiomyoma(UL).METHODS:UL was induced in rats using exogenous estrogen and progesterone.LD was administered(p.o.) for 4 weeks,and mifepristone(RU-486)used as a control.To observe the effect of LD on the uterine coefficient and uterine transverse diameter,a radioimmunoassay method was used to detect serum levels of sex hormones.Light microscopic analyses of pathologic changes in the tissues of UL rats were evaluated.Expression of the proteins of matrix metalloproteinases(MMPs) and tissue inhibitors of metalloproteinases(TIMPs) in uterine tissues was assessed by immunohistochemical staining and western blotting.RESULTS:A UL model in rats was established successfully.LD reduced uterine weight,uterine coefficient,and uterine transverse diameter compared with untreated controls.LD reduced levels of estradiol,progesterone,follicle-stimulating hormone,and luteinizing hormone in our UL models.LD improved the pathologic condition of uterine muscle.Expression of MMP-2 protein decreased to varying extents in LD-treated groups,but TIMP-2 levels were enhanced.LD appears to reduce MMP-2 expression and increase TIMP-2 expression in UL tissue.CONCLUSION:These data suggest that the mechanism of action of LD on ULs may involve reduction of MMP-2 expression and increase in TIMP-2 expression in rats.展开更多
OBJECTIVE:To investigate effects of Saikosaponin D(SSd) on syndecan-2,matrix metalloproteinases(MMPs) and tissue inhibitor of metalloproteinases-2(TIMP-2) in livers of rat with hepatocellular carcinoma(HCC).METHODS:Ma...OBJECTIVE:To investigate effects of Saikosaponin D(SSd) on syndecan-2,matrix metalloproteinases(MMPs) and tissue inhibitor of metalloproteinases-2(TIMP-2) in livers of rat with hepatocellular carcinoma(HCC).METHODS:Male SD rats were divided into control(n=10),model(n=20) and SSd(n=20) groups,and model and SSd groups given intragastric 0.2%(w/v) N-diethylnitrosamine to induce HCC.SSd group received 0.03%(w/v) SSd in saline.Liver samples were analysed immunohistochemically for syndecan-2,MMP-2,MMP-13 and TIMP-2 at 16 weeks.RESULTS:The model group had more malignant nodules than the SSd group;all model-group HCC cells were grade III;SSd-group HCC cells were grades I-II.Controls showed normal hepatic cell phenotypes and no syndecan-2 + staining.Syndecan-2 + staining was greater in the model group(35.2%,P≤0.001) than in controls or the SSd group(16.5%,P ≤ 0.001).The model group had more intense MMP-2 + staining than controls(0.37 vs 0.27,P≤0.01) or the SSd group(0.31 vs 0.37,P≤0.05);and higher MMP-13 + staining(72.55%) than in controls(12.55%,P≤0.001) and SSd group(20.18%,P≤0.01).The model group also had more TIMP-2 + staining(57.2%) than controls(20.9%,P≤0.001) and SSd group(22.7%,P≤0.001).Controls and SSd group showed no difference in TIMP-2 + rates.CONCLUSION:SSd inhibited HCC development,and downregulated expression of syndecan-2,MMP-2,MMP-13 and TIMP-2 in rat HCC liver tissue.展开更多
In this study, genes of two distinct tissue inhibitors of metalloproteinases-2 (TIMP-2) from Japanese puffer fishFugu rubripes, Fugu TIMP-2a and TIMP-2b, were cloned. The open reading frames of Fugu TIMP-2a and TIMP-2...In this study, genes of two distinct tissue inhibitors of metalloproteinases-2 (TIMP-2) from Japanese puffer fishFugu rubripes, Fugu TIMP-2a and TIMP-2b, were cloned. The open reading frames of Fugu TIMP-2a and TIMP-2b cDNAsare composed of 660 and 657 nucleotides and 220 and 219 amino acids, respectively. Both Fugu TIMP-2s contain 12 cysteineresidues, which might form six disulfide bonds as in other animals’ TIMP-2s. Reverse-transcribed polymerase chain reactionanalysis showed the mRNAs of Fugu TIMP-2a and TIMP-2b to be expressed in some tissues examined with different expres-sion patterns. These findings suggest that the two distinct Fugu TIMP-2s might perform different functions in Fugu tissues.展开更多
AIM:To examine the expressions of matrix metalloproteinases-2(MMP-2)and tissue inhibitor of metalloproteinases-1(TINP-1)in rat fibrotic liver and in normal rat hepatic stellate cells,and to investigate the changes in ...AIM:To examine the expressions of matrix metalloproteinases-2(MMP-2)and tissue inhibitor of metalloproteinases-1(TINP-1)in rat fibrotic liver and in normal rat hepatic stellate cells,and to investigate the changes in their expressions in response to treatment with interleukin-10(IL-10)and platelet-derived growth factor(PDGF).METHODS:Rat models of CCl4-induced hepatic fibrosis were established and the liver tissues were sampled from the rats with or without IL-10 treatment,and also from the control rats.The expressions of MMP-2 and TIMP-1 in liver tissues were detected by S-P immunohistochemistry,and their expression intensities were evaluated in different groups.Hepatic stellate cells(HSCs)were isolated from normal rat and cultured in vitro prior to exposure to PDGF treatment or co-treatment with IL-10 and PDGF.MMP-2 and TIMP-1 levels were measured by semi-quantitative reverse transcriptional polymerase chain reaction(RT-PCR).RESULTS:CCl4-induced rat hepatic fibrosis models were successfully established.The positive expressions of MMP-2 and TIMP-1 increased obviously with the development of hepatic fibrosis,especially in untreated model group(84.0%and 92.0%,P<0.01).The positive signals decreased significantly following IL-10 treatment(39.3%and 71.4%,P<0.01 and P<0.05)in a time-dependent manner.TIMP-1 mRNA in PDGF-treated group was significantly increased time-dependently in comparison with that of the control group,but PDGF did not obviously affect MMP-2 expression.No difference was noted in TIMP-1 and MMP-2 expressions in HSCs after IL-10 and PDGF treatment(P>0.05).CONCLUSION:MMP-2 and TIMP-1 expressions increase in liver tissues with the development of fibrosis,which can be inhibited by exogenous IL-10 inhibitor.PDGF induces the up-regulation of TIMP-1 but not MMP-2 in the HSCs.IL-10 inhibits TIMP-1 and MMP-2 expressions in HSCs induced by PDGF.展开更多
基金Supported by National Natural Science Fund(Study on the Action Mechanism of Inhibition of Uterine Leiomyoma of Regulation of Lichong Decoction for Ang-Tie-2 Transduction Pathway,No.81373812Research on the Regulating Mechanism of the Inhibitory Effect of Nourishing Healthy Qi and Eliminating Blood Stasis Chinese Medicine on Extracellular Matrix Metabolize in Uterine Leiomyoma,No.81073096)
文摘OBJECTIVE:To study the effect of Lichong decoction(LD) on expression of matrix metalloproteinase-2(MMP-2) and metalloproteinase-2(TIMP-2) in a rat model of uterine leiomyoma(UL).METHODS:UL was induced in rats using exogenous estrogen and progesterone.LD was administered(p.o.) for 4 weeks,and mifepristone(RU-486)used as a control.To observe the effect of LD on the uterine coefficient and uterine transverse diameter,a radioimmunoassay method was used to detect serum levels of sex hormones.Light microscopic analyses of pathologic changes in the tissues of UL rats were evaluated.Expression of the proteins of matrix metalloproteinases(MMPs) and tissue inhibitors of metalloproteinases(TIMPs) in uterine tissues was assessed by immunohistochemical staining and western blotting.RESULTS:A UL model in rats was established successfully.LD reduced uterine weight,uterine coefficient,and uterine transverse diameter compared with untreated controls.LD reduced levels of estradiol,progesterone,follicle-stimulating hormone,and luteinizing hormone in our UL models.LD improved the pathologic condition of uterine muscle.Expression of MMP-2 protein decreased to varying extents in LD-treated groups,but TIMP-2 levels were enhanced.LD appears to reduce MMP-2 expression and increase TIMP-2 expression in UL tissue.CONCLUSION:These data suggest that the mechanism of action of LD on ULs may involve reduction of MMP-2 expression and increase in TIMP-2 expression in rats.
基金Supported by the National Natural Science Foundation of China(No.30471982)
文摘OBJECTIVE:To investigate effects of Saikosaponin D(SSd) on syndecan-2,matrix metalloproteinases(MMPs) and tissue inhibitor of metalloproteinases-2(TIMP-2) in livers of rat with hepatocellular carcinoma(HCC).METHODS:Male SD rats were divided into control(n=10),model(n=20) and SSd(n=20) groups,and model and SSd groups given intragastric 0.2%(w/v) N-diethylnitrosamine to induce HCC.SSd group received 0.03%(w/v) SSd in saline.Liver samples were analysed immunohistochemically for syndecan-2,MMP-2,MMP-13 and TIMP-2 at 16 weeks.RESULTS:The model group had more malignant nodules than the SSd group;all model-group HCC cells were grade III;SSd-group HCC cells were grades I-II.Controls showed normal hepatic cell phenotypes and no syndecan-2 + staining.Syndecan-2 + staining was greater in the model group(35.2%,P≤0.001) than in controls or the SSd group(16.5%,P ≤ 0.001).The model group had more intense MMP-2 + staining than controls(0.37 vs 0.27,P≤0.01) or the SSd group(0.31 vs 0.37,P≤0.05);and higher MMP-13 + staining(72.55%) than in controls(12.55%,P≤0.001) and SSd group(20.18%,P≤0.01).The model group also had more TIMP-2 + staining(57.2%) than controls(20.9%,P≤0.001) and SSd group(22.7%,P≤0.001).Controls and SSd group showed no difference in TIMP-2 + rates.CONCLUSION:SSd inhibited HCC development,and downregulated expression of syndecan-2,MMP-2,MMP-13 and TIMP-2 in rat HCC liver tissue.
基金partially supported by a grant from the Ministry of Education,Culture,Sports,Science and Technology of Japan.
文摘In this study, genes of two distinct tissue inhibitors of metalloproteinases-2 (TIMP-2) from Japanese puffer fishFugu rubripes, Fugu TIMP-2a and TIMP-2b, were cloned. The open reading frames of Fugu TIMP-2a and TIMP-2b cDNAsare composed of 660 and 657 nucleotides and 220 and 219 amino acids, respectively. Both Fugu TIMP-2s contain 12 cysteineresidues, which might form six disulfide bonds as in other animals’ TIMP-2s. Reverse-transcribed polymerase chain reactionanalysis showed the mRNAs of Fugu TIMP-2a and TIMP-2b to be expressed in some tissues examined with different expres-sion patterns. These findings suggest that the two distinct Fugu TIMP-2s might perform different functions in Fugu tissues.
基金Supported by the Science and Technology Fund of Fujian Province,No.2003D05
文摘AIM:To examine the expressions of matrix metalloproteinases-2(MMP-2)and tissue inhibitor of metalloproteinases-1(TINP-1)in rat fibrotic liver and in normal rat hepatic stellate cells,and to investigate the changes in their expressions in response to treatment with interleukin-10(IL-10)and platelet-derived growth factor(PDGF).METHODS:Rat models of CCl4-induced hepatic fibrosis were established and the liver tissues were sampled from the rats with or without IL-10 treatment,and also from the control rats.The expressions of MMP-2 and TIMP-1 in liver tissues were detected by S-P immunohistochemistry,and their expression intensities were evaluated in different groups.Hepatic stellate cells(HSCs)were isolated from normal rat and cultured in vitro prior to exposure to PDGF treatment or co-treatment with IL-10 and PDGF.MMP-2 and TIMP-1 levels were measured by semi-quantitative reverse transcriptional polymerase chain reaction(RT-PCR).RESULTS:CCl4-induced rat hepatic fibrosis models were successfully established.The positive expressions of MMP-2 and TIMP-1 increased obviously with the development of hepatic fibrosis,especially in untreated model group(84.0%and 92.0%,P<0.01).The positive signals decreased significantly following IL-10 treatment(39.3%and 71.4%,P<0.01 and P<0.05)in a time-dependent manner.TIMP-1 mRNA in PDGF-treated group was significantly increased time-dependently in comparison with that of the control group,but PDGF did not obviously affect MMP-2 expression.No difference was noted in TIMP-1 and MMP-2 expressions in HSCs after IL-10 and PDGF treatment(P>0.05).CONCLUSION:MMP-2 and TIMP-1 expressions increase in liver tissues with the development of fibrosis,which can be inhibited by exogenous IL-10 inhibitor.PDGF induces the up-regulation of TIMP-1 but not MMP-2 in the HSCs.IL-10 inhibits TIMP-1 and MMP-2 expressions in HSCs induced by PDGF.
