Objective: To elucidate the relation between human tissue factor pathwayinhibitor-2 (TFPI-2) expression and ovarian tumor migration and invasion. Methods: Human TFPI-2expression vector pBos-Cite-neo/TFPI-2 was transfe...Objective: To elucidate the relation between human tissue factor pathwayinhibitor-2 (TFPI-2) expression and ovarian tumor migration and invasion. Methods: Human TFPI-2expression vector pBos-Cite-neo/TFPI-2 was transfected into ovarian tumor cells line A2780- Afterthe transfected cells were selected by G418, transfected and nontransfected cells were screened forTFPI-2 mRNA and protein by reverse transcription-polymerase chain reaction and Western blotanalysis, respectively. The number of transfected or nontransfected cells passing through membraneof Boyden chamber was counted as the basis assessing tumor cells migratory and invasive behaviors.Results: Expression of mRNA and protein of TFPI-2 was detectable in transfected cells. In invasionassay, the number of TFPI-2-expressing cells to traverse a Matrigel-coated membrane was obviouslydecreased compared with that of nonexpressing cells (59.3±6.5 vs 109.7±5.5, P 【 0.01); While inmigration assay, no significant difference through a noncoated membrane was observed amongtransfected and nontransfected cells (114.7±8.6 vs 127.3±7.1, P 】 0.05). Conclusion: Expression ofTFPI-2 may strongly inhibit the invasive ability of ovarian tumor cells in vitro, but has no effecton the migratory ability which provides an experimental basis for genotherapy of human ovariantumor.展开更多
This study examined the expressions of human serum tissue factor （TF） and tissue factor pathway inhibitor （TFPI） in patients with acute graft-versus-host disease （aGVHD） after allogeneic hematopoietic stem cell ...This study examined the expressions of human serum tissue factor （TF） and tissue factor pathway inhibitor （TFPI） in patients with acute graft-versus-host disease （aGVHD） after allogeneic hematopoietic stem cell transplantation （allo-HSCT） and their clinical significance. The serum TF and TFPI levels were detected by ELISA in 28 allo-HSCT recipients before and after the transplanta-tion and the changes of TF and TFPI levels were dynamically monitored at different phases of the disease. No significant differences in the serum TF and TFPI levels were found in allo-HSCT recipi-ents in the absence of aGVHD or with gradeⅠaGVHD before and after the transplantation. The lev-els of serum TF and TFPI were substantially increased in the patients with gradeⅡ aGVHD at the peak of aGVHD （P〈0.05） and they were even higher in the patients with grade Ⅲ–Ⅳ aGVHD （P〈0.01）. When the conditions became stable after treatment with immunosuppressive agents, the serum TFPI level was decreased to the baseline level （P〉0.05） and the TF level was lowered but still higher than the baseline level （P〈0.05）. It was concluded that the levels of serum TF and TFPI were increased significantly in the patients with grade Ⅱ–Ⅳ aGVHD after allo-HSCT and decreased markedly after the treatment. Monitoring the levels of serum TF and TFPI in the patients with allo-HSCT is important to predict the occurrence, outcome and prognosis of aGVHD.展开更多
AIM: To investigate the clinical significance of expression of tissue factor (TF) and tissue factor pathway inhibitor (TFPI) in ulcerative colitis (UC).
We have observed earlier that testosterone at physiological concentrations can stimulate tissue factor pathway inhibitor(TFPI)gene expression through the androgen receptor in endothelial cells.This study further inves...We have observed earlier that testosterone at physiological concentrations can stimulate tissue factor pathway inhibitor(TFPI)gene expression through the androgen receptor in endothelial cells.This study further investigated the impact of testosterone on TFPI levels in response to inflammatory cytokine tumor necrosis factor-alpha(TNF-α).Cultured human umbilical vein endothelial cells were incubated in the presence or absence of testosterone or TNF-α.TFPI protein and mRNA levels were assessed by enzyme-linked immunosorbent assay and quantitative real-time reverse transcription polymerase chain reaction.To study the cellular mechanism of testosterone’s action,nuclear factor-kappa B(NF-κB)translocation was confirmed by electrophoretic mobility shift assays.We found that after NF-κB was activated by TNF-α,TFPI protein levels declined significantly by 37.3%compared with controls(P<0.001),and the mRNA levels of TFPI also decreased greatly(P<0.001).A concentration of 30 nmol L-1 testosterone increased the secretion of TFPI compared with the TNF-α-treated group.NF-κB DNA-binding activity was significantly suppressed by testosterone(P<0.05).This suggests that physiological testosterone concentrations may exert their antithrombotic effects on TFPI expression during inflammation by downregulating NF-κB activity.展开更多
Transcription factors(TFs)orchestrate the regulation of cellular gene expression and thereby determine cell functionality.In this study,we analyzed the distribution of TFs containing domains,which named as ZnFTFs,both...Transcription factors(TFs)orchestrate the regulation of cellular gene expression and thereby determine cell functionality.In this study,we analyzed the distribution of TFs containing domains,which named as ZnFTFs,both in ascomycete and basidiomycete fungi.We found that ZnFTFs were widely distributed in these fungal species,but there was more expansion of the ZnFTF class in Ascomycota than Basidiomycota.We identified 40 ZnFTFs in Ustilaginoidea virens,and demonstrated the involvement of UvZnFTF1 in vegetative growth,conidiation,pigment biosynthesis and pathogenicity.RNA-Seq analysis suggested that UvZnFTF1 may regulate different nutrient metabolism pathways,the production of secondary metabolites,and the expression of pathogen-host interaction genes and secreted protein-encodi ng genes.Analysis of the distributi on of differe nt fungal TFs in U.virens further dem on strated that UvZnFTFs make up a large TF family and may play essential biological roles in U.virens.展开更多
The relationship between angiogenesis and eukaryotic translation initiation factor 4E （EIF4E） expression level in non Hodgkin lymphoma （NHL） was studied. Mean microvessel density （MVD） and EIF4E were detected in...The relationship between angiogenesis and eukaryotic translation initiation factor 4E （EIF4E） expression level in non Hodgkin lymphoma （NHL） was studied. Mean microvessel density （MVD） and EIF4E were detected in 52 lymph node samples paraffin sections of patients with newly diagnosed NHL by the way of immunohistochemistry. Antisense EIF4E cDNA was cloned into plasmid pcDNA3.1 （＋） and transfected into Raji cells. A series of angiogenesis related factors,including vascular endothelial growth factor （VEGF）, matrix metalloproteinases 9 （MMP-9） and tissue inhibitor of metalloproteinases-2 （TIMP-2） proteins were detected by Western blot. The results showed that： （1） The Expression of EIF4E and MVD was higher in aggressive lymphomas than in indolent lymphomas（P〈0.05）and the expression of EIF4E was positively correlated with MVD in lymph node of NHL（r=0. 695, P〈0.01）. （2） Antisense EIF4E eukaryocytic expression vector （pcDNA3. 1-EIF4Eas） was constructed successfully. （3） EIF4E, VEGF and MMP-9 were expressed at high levels in Raji cells as compared to normal human peripheral blood monocular cells （NHPMC）, and blockage of EIF4E expression brought down the expression of VEGF and MMP-9. However, TIMP-2 was undetectable in Rail cells, although a moderate level of TIMP-2 was detected in NHPMC. It was concluded that the increased EIF4E expression was associated with aggressive property of NHL.展开更多
Objective: To fractionate and identify polyphenols from Guazuma ulmifolia Lam. leaves, and to explore their antioxidant, 5-hydroxy-3-methylglutaryl-coenzyme A(HMG-Co A) reductase inhibitory, and Nrf2 modulatory activi...Objective: To fractionate and identify polyphenols from Guazuma ulmifolia Lam. leaves, and to explore their antioxidant, 5-hydroxy-3-methylglutaryl-coenzyme A(HMG-Co A) reductase inhibitory, and Nrf2 modulatory activities.Methods: The 1,1-diphenyl-2-picrylhydrazyl assay was used to evaluate the antioxidant activity of a polyphenolic fraction of the extract of Guazuma ulmifolia Lam. leaves. THP-1 gene reporter cell lines constructed with a transcriptional response element specific for Nrf2 and a minimal promoter for the firefly luciferase–green fluorescent protein transgene were used to determine the effect of the polyphenolic fraction on the Nrf2 signaling pathway. Furthermore, an assay of HMG-Co A reductase inhibitory activity was performed by using a commercial enzyme kit. Polyphenolic compounds were identified by liquid chromatographytandem mass spectrometry.Results: The polyphenolic fraction showed fairly strong antioxidant activity [IC50 =(14.90 ± 4.70) μg/m L] and inhibited HMG-Co A reductase activity by 69.10%, which was slightly lower than that by pravastatin(84.37%) and quercetin(84.25%). Additionally, the polyphenolic fraction activated the Nrf2 antioxidant signaling pathway at 500 μg/m L. Eleven subfractions resulting from the column chromatography separation of the polyphenolic fraction also showed relatively strong antioxidant activities(IC50: 17.46–217.14 μg/m L). The subfraction(F6) stimulated the Nrf2 signaling pathway and had HMG-Co A reductase inhibitory activity(65.43%). Moreover, the subfraction contained two main flavonoids: quercetin and quercimeritrin.Conclusions: The polyphenolic fraction of Guazuma ulmifolia could induce antioxidant genes via the Nrf2/antioxidant regulatory elements pathway, and is a promising candidate for an inhibitor of HMG-Co A reductase.展开更多
Objective To establish the correlation between tissue factor pathway inhibitor 2 (TFPI-2) and the progression of pancreatic carcinoma(PC) after detecting the expression level of TFPI-2 in PC,and to evaluate the value ...Objective To establish the correlation between tissue factor pathway inhibitor 2 (TFPI-2) and the progression of pancreatic carcinoma(PC) after detecting the expression level of TFPI-2 in PC,and to evaluate the value of TFPI-2 as prognostic index in PC.Methods Expression levels of TFPI-2 in 10 normal and 25 cancerous/juxta-cancerous pancreas were testified with Western blot and RTPCR analyses respectively.Expression density of TFPI-2 in each group was analyzed with HPIAS image analyzer and compared with ANOVA.The correlation between the expression level of TFPI-2 and malignancy was tested with Spearman rank correlation.Disease-specific survival curves were calculated according to Kaplan Meier algorithm,and log rank test was used to compare survival curves.Then,Cox regression analysis was applied to determine the single contribution of each covariate on survival rate.Results The expression level of TFPI-2 decreased along with progression of PC with significant difference among groups (P <0.05),and there was a significantly negative correlation between TFPI-2 protein and progression (r= -ft 816,P <0.001).Among the 13 studied variables,such as the expression level of TFPI-2 protein, tumor stage,portal vein resection,lymph node metastasis,only lymph node metastasis was a predictor of outcome.However,when we analyzed the survival without considering lymph node metastasis,a stepwise Cox analysis showed that expression level of TFPI-2 protein and combined organ resection were significantly associated to survival.Conclusion The data showed that there was strongly correlation between the expression level of TFPI-2 and the progression and survival of PC,which suggested that TFPI-2 could be a newly prognostic factor and a novel approach for gene therapy for PC.展开更多
目的观察高糖刺激的大鼠肾小球系膜细胞基质金属蛋白酶-2(MMP-2)及其组织抑制物-2(TIMP-2)、膜型基质金属蛋白酶-1(MT1-MMP)和结缔组织生长因子(CTGF)的动态变化以探讨糖尿病肾病(DN)的发病机制。方法体外培养的大鼠HBZY-1肾小球系膜细...目的观察高糖刺激的大鼠肾小球系膜细胞基质金属蛋白酶-2(MMP-2)及其组织抑制物-2(TIMP-2)、膜型基质金属蛋白酶-1(MT1-MMP)和结缔组织生长因子(CTGF)的动态变化以探讨糖尿病肾病(DN)的发病机制。方法体外培养的大鼠HBZY-1肾小球系膜细胞分为低糖(5.5mmol/L葡萄糖)组、高糖(30mmol/L葡萄糖)组和渗透压对照(5.5mmol/L葡萄糖+24.5mmol/L甘露醇)组,24、48、72、96h后采用RT-PCR及Western blotting法分别检测MMP-2、TIMP-2、MT1-MMP及CTGF的mRNA及蛋白表达情况,酶联免疫吸附法(ELISA)检测培养上清中Ⅳ型胶原的含量。结果 Western blotting结果显示,与低糖组相比,高糖组MMP-2的表达在24h时略有升高,较低糖组增加10%±4%(P<0.05),至48h时则较低糖组减少42%±2%,其后随时间延长表达持续降低,至96h时较低糖组减少78%±2%;MT1-MMP表达在24h开始下降并随时间呈下降趋势,与低糖组相比较,在刺激的24h,高糖组MT1-MMP表达下降了29%±3%,随后持续下降,至96h则下降了78%±9%(P<0.01)。高糖组各时间点TIMP-2和CTGF表达均较低糖组增高,其中CTGF的表达在高糖刺激的24h即显著增高,为低糖组的201%±24%,随后持续增高,至培养96h为低糖组的484%±51%(P<0.01);TIMP-2的表达在24h时较低糖组增加55%±3%,且随时间延长呈增高趋势(P<0.01)。MMP-2、TIMP-2、MT1-MMP和CTGF的mRNA表达与相应蛋白的表达趋势基本一致。与低糖组相比,高糖组细胞上清中的Ⅳ型胶原于24h即有增加,且持续增高至96h(P<0.05)。低糖组和渗透压对照组组内、组间的各指标差异均无统计学意义。结论尽管高糖刺激早期可小幅诱导MMP-2的表达增强,但长期高糖刺激则可抑制MMP-2和MT1-MMP的表达及活化,同时促进系膜细胞TIMP-2和CTGF的表达。DN中肾小球细胞外基质的积聚可能是由于上述细胞因子和蛋白酶引起细胞外基质代谢失衡所致。展开更多
文摘Objective: To elucidate the relation between human tissue factor pathwayinhibitor-2 (TFPI-2) expression and ovarian tumor migration and invasion. Methods: Human TFPI-2expression vector pBos-Cite-neo/TFPI-2 was transfected into ovarian tumor cells line A2780- Afterthe transfected cells were selected by G418, transfected and nontransfected cells were screened forTFPI-2 mRNA and protein by reverse transcription-polymerase chain reaction and Western blotanalysis, respectively. The number of transfected or nontransfected cells passing through membraneof Boyden chamber was counted as the basis assessing tumor cells migratory and invasive behaviors.Results: Expression of mRNA and protein of TFPI-2 was detectable in transfected cells. In invasionassay, the number of TFPI-2-expressing cells to traverse a Matrigel-coated membrane was obviouslydecreased compared with that of nonexpressing cells (59.3±6.5 vs 109.7±5.5, P 【 0.01); While inmigration assay, no significant difference through a noncoated membrane was observed amongtransfected and nontransfected cells (114.7±8.6 vs 127.3±7.1, P 】 0.05). Conclusion: Expression ofTFPI-2 may strongly inhibit the invasive ability of ovarian tumor cells in vitro, but has no effecton the migratory ability which provides an experimental basis for genotherapy of human ovariantumor.
文摘This study examined the expressions of human serum tissue factor （TF） and tissue factor pathway inhibitor （TFPI） in patients with acute graft-versus-host disease （aGVHD） after allogeneic hematopoietic stem cell transplantation （allo-HSCT） and their clinical significance. The serum TF and TFPI levels were detected by ELISA in 28 allo-HSCT recipients before and after the transplanta-tion and the changes of TF and TFPI levels were dynamically monitored at different phases of the disease. No significant differences in the serum TF and TFPI levels were found in allo-HSCT recipi-ents in the absence of aGVHD or with gradeⅠaGVHD before and after the transplantation. The lev-els of serum TF and TFPI were substantially increased in the patients with gradeⅡ aGVHD at the peak of aGVHD （P〈0.05） and they were even higher in the patients with grade Ⅲ–Ⅳ aGVHD （P〈0.01）. When the conditions became stable after treatment with immunosuppressive agents, the serum TFPI level was decreased to the baseline level （P〉0.05） and the TF level was lowered but still higher than the baseline level （P〈0.05）. It was concluded that the levels of serum TF and TFPI were increased significantly in the patients with grade Ⅱ–Ⅳ aGVHD after allo-HSCT and decreased markedly after the treatment. Monitoring the levels of serum TF and TFPI in the patients with allo-HSCT is important to predict the occurrence, outcome and prognosis of aGVHD.
文摘AIM: To investigate the clinical significance of expression of tissue factor (TF) and tissue factor pathway inhibitor (TFPI) in ulcerative colitis (UC).
基金the National Natural Science Foundation of China(No.30670842)the Natural Science Foundation of Guangdong Province,China(No.5300582).
文摘We have observed earlier that testosterone at physiological concentrations can stimulate tissue factor pathway inhibitor(TFPI)gene expression through the androgen receptor in endothelial cells.This study further investigated the impact of testosterone on TFPI levels in response to inflammatory cytokine tumor necrosis factor-alpha(TNF-α).Cultured human umbilical vein endothelial cells were incubated in the presence or absence of testosterone or TNF-α.TFPI protein and mRNA levels were assessed by enzyme-linked immunosorbent assay and quantitative real-time reverse transcription polymerase chain reaction.To study the cellular mechanism of testosterone’s action,nuclear factor-kappa B(NF-κB)translocation was confirmed by electrophoretic mobility shift assays.We found that after NF-κB was activated by TNF-α,TFPI protein levels declined significantly by 37.3%compared with controls(P<0.001),and the mRNA levels of TFPI also decreased greatly(P<0.001).A concentration of 30 nmol L-1 testosterone increased the secretion of TFPI compared with the TNF-α-treated group.NF-κB DNA-binding activity was significantly suppressed by testosterone(P<0.05).This suggests that physiological testosterone concentrations may exert their antithrombotic effects on TFPI expression during inflammation by downregulating NF-κB activity.
基金supported by the National Natural Science Foundation of China(Grant No.31601593)the Young Elite Scientist Sponsorship of China Association for Science and Technology(Grant No.YESS20170108)the Natural Science Foundation of Jiangsu Province,China(Grant No.BK20160588).
文摘Transcription factors(TFs)orchestrate the regulation of cellular gene expression and thereby determine cell functionality.In this study,we analyzed the distribution of TFs containing domains,which named as ZnFTFs,both in ascomycete and basidiomycete fungi.We found that ZnFTFs were widely distributed in these fungal species,but there was more expansion of the ZnFTF class in Ascomycota than Basidiomycota.We identified 40 ZnFTFs in Ustilaginoidea virens,and demonstrated the involvement of UvZnFTF1 in vegetative growth,conidiation,pigment biosynthesis and pathogenicity.RNA-Seq analysis suggested that UvZnFTF1 may regulate different nutrient metabolism pathways,the production of secondary metabolites,and the expression of pathogen-host interaction genes and secreted protein-encodi ng genes.Analysis of the distributi on of differe nt fungal TFs in U.virens further dem on strated that UvZnFTFs make up a large TF family and may play essential biological roles in U.virens.
文摘The relationship between angiogenesis and eukaryotic translation initiation factor 4E （EIF4E） expression level in non Hodgkin lymphoma （NHL） was studied. Mean microvessel density （MVD） and EIF4E were detected in 52 lymph node samples paraffin sections of patients with newly diagnosed NHL by the way of immunohistochemistry. Antisense EIF4E cDNA was cloned into plasmid pcDNA3.1 （＋） and transfected into Raji cells. A series of angiogenesis related factors,including vascular endothelial growth factor （VEGF）, matrix metalloproteinases 9 （MMP-9） and tissue inhibitor of metalloproteinases-2 （TIMP-2） proteins were detected by Western blot. The results showed that： （1） The Expression of EIF4E and MVD was higher in aggressive lymphomas than in indolent lymphomas（P〈0.05）and the expression of EIF4E was positively correlated with MVD in lymph node of NHL（r=0. 695, P〈0.01）. （2） Antisense EIF4E eukaryocytic expression vector （pcDNA3. 1-EIF4Eas） was constructed successfully. （3） EIF4E, VEGF and MMP-9 were expressed at high levels in Raji cells as compared to normal human peripheral blood monocular cells （NHPMC）, and blockage of EIF4E expression brought down the expression of VEGF and MMP-9. However, TIMP-2 was undetectable in Rail cells, although a moderate level of TIMP-2 was detected in NHPMC. It was concluded that the increased EIF4E expression was associated with aggressive property of NHL.
基金funded by International Collaboration Research Grant under the Ministry of Research, Technology and Higher Education of the Republic of Indonesia (No. 011/SP2H/LT/DRPM/VIII/2017)University of Missouri-Columbia, MO(USA)
文摘Objective: To fractionate and identify polyphenols from Guazuma ulmifolia Lam. leaves, and to explore their antioxidant, 5-hydroxy-3-methylglutaryl-coenzyme A(HMG-Co A) reductase inhibitory, and Nrf2 modulatory activities.Methods: The 1,1-diphenyl-2-picrylhydrazyl assay was used to evaluate the antioxidant activity of a polyphenolic fraction of the extract of Guazuma ulmifolia Lam. leaves. THP-1 gene reporter cell lines constructed with a transcriptional response element specific for Nrf2 and a minimal promoter for the firefly luciferase–green fluorescent protein transgene were used to determine the effect of the polyphenolic fraction on the Nrf2 signaling pathway. Furthermore, an assay of HMG-Co A reductase inhibitory activity was performed by using a commercial enzyme kit. Polyphenolic compounds were identified by liquid chromatographytandem mass spectrometry.Results: The polyphenolic fraction showed fairly strong antioxidant activity [IC50 =(14.90 ± 4.70) μg/m L] and inhibited HMG-Co A reductase activity by 69.10%, which was slightly lower than that by pravastatin(84.37%) and quercetin(84.25%). Additionally, the polyphenolic fraction activated the Nrf2 antioxidant signaling pathway at 500 μg/m L. Eleven subfractions resulting from the column chromatography separation of the polyphenolic fraction also showed relatively strong antioxidant activities(IC50: 17.46–217.14 μg/m L). The subfraction(F6) stimulated the Nrf2 signaling pathway and had HMG-Co A reductase inhibitory activity(65.43%). Moreover, the subfraction contained two main flavonoids: quercetin and quercimeritrin.Conclusions: The polyphenolic fraction of Guazuma ulmifolia could induce antioxidant genes via the Nrf2/antioxidant regulatory elements pathway, and is a promising candidate for an inhibitor of HMG-Co A reductase.
基金Supported by the National Natural Science Foundation of China(81100664)Wuhan University Independent Research Project(111091)
文摘Objective To establish the correlation between tissue factor pathway inhibitor 2 (TFPI-2) and the progression of pancreatic carcinoma(PC) after detecting the expression level of TFPI-2 in PC,and to evaluate the value of TFPI-2 as prognostic index in PC.Methods Expression levels of TFPI-2 in 10 normal and 25 cancerous/juxta-cancerous pancreas were testified with Western blot and RTPCR analyses respectively.Expression density of TFPI-2 in each group was analyzed with HPIAS image analyzer and compared with ANOVA.The correlation between the expression level of TFPI-2 and malignancy was tested with Spearman rank correlation.Disease-specific survival curves were calculated according to Kaplan Meier algorithm,and log rank test was used to compare survival curves.Then,Cox regression analysis was applied to determine the single contribution of each covariate on survival rate.Results The expression level of TFPI-2 decreased along with progression of PC with significant difference among groups (P <0.05),and there was a significantly negative correlation between TFPI-2 protein and progression (r= -ft 816,P <0.001).Among the 13 studied variables,such as the expression level of TFPI-2 protein, tumor stage,portal vein resection,lymph node metastasis,only lymph node metastasis was a predictor of outcome.However,when we analyzed the survival without considering lymph node metastasis,a stepwise Cox analysis showed that expression level of TFPI-2 protein and combined organ resection were significantly associated to survival.Conclusion The data showed that there was strongly correlation between the expression level of TFPI-2 and the progression and survival of PC,which suggested that TFPI-2 could be a newly prognostic factor and a novel approach for gene therapy for PC.
文摘目的观察高糖刺激的大鼠肾小球系膜细胞基质金属蛋白酶-2(MMP-2)及其组织抑制物-2(TIMP-2)、膜型基质金属蛋白酶-1(MT1-MMP)和结缔组织生长因子(CTGF)的动态变化以探讨糖尿病肾病(DN)的发病机制。方法体外培养的大鼠HBZY-1肾小球系膜细胞分为低糖(5.5mmol/L葡萄糖)组、高糖(30mmol/L葡萄糖)组和渗透压对照(5.5mmol/L葡萄糖+24.5mmol/L甘露醇)组,24、48、72、96h后采用RT-PCR及Western blotting法分别检测MMP-2、TIMP-2、MT1-MMP及CTGF的mRNA及蛋白表达情况,酶联免疫吸附法(ELISA)检测培养上清中Ⅳ型胶原的含量。结果 Western blotting结果显示,与低糖组相比,高糖组MMP-2的表达在24h时略有升高,较低糖组增加10%±4%(P<0.05),至48h时则较低糖组减少42%±2%,其后随时间延长表达持续降低,至96h时较低糖组减少78%±2%;MT1-MMP表达在24h开始下降并随时间呈下降趋势,与低糖组相比较,在刺激的24h,高糖组MT1-MMP表达下降了29%±3%,随后持续下降,至96h则下降了78%±9%(P<0.01)。高糖组各时间点TIMP-2和CTGF表达均较低糖组增高,其中CTGF的表达在高糖刺激的24h即显著增高,为低糖组的201%±24%,随后持续增高,至培养96h为低糖组的484%±51%(P<0.01);TIMP-2的表达在24h时较低糖组增加55%±3%,且随时间延长呈增高趋势(P<0.01)。MMP-2、TIMP-2、MT1-MMP和CTGF的mRNA表达与相应蛋白的表达趋势基本一致。与低糖组相比,高糖组细胞上清中的Ⅳ型胶原于24h即有增加,且持续增高至96h(P<0.05)。低糖组和渗透压对照组组内、组间的各指标差异均无统计学意义。结论尽管高糖刺激早期可小幅诱导MMP-2的表达增强,但长期高糖刺激则可抑制MMP-2和MT1-MMP的表达及活化,同时促进系膜细胞TIMP-2和CTGF的表达。DN中肾小球细胞外基质的积聚可能是由于上述细胞因子和蛋白酶引起细胞外基质代谢失衡所致。
