Obesity is a major human health problem associated with various diseases, including cardiac injury and type 2 diabetes. Trapa japonica Flerov (TJF) has been used in traditional oriental medicine to treat diabetes. In ...Obesity is a major human health problem associated with various diseases, including cardiac injury and type 2 diabetes. Trapa japonica Flerov (TJF) has been used in traditional oriental medicine to treat diabetes. In this study, we evaluated the inhibitory effect of and the mechanism underlying the effect of TJF extract on adipogenesis in 3T3-L1 cells. The effects of TJF extract on cell viability were analyzed using a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay, and the anti-adipogenic effect was measured by oil red O staining. The expression of peroxisomal proliferator activated receptor (PPAR)γ, CCAAT/enhancer-binding protein-α (C/EBP)α, adenosine monophosphate-activated protein kinase (AMPK), acetyl-CoA carboxylase (ACC), adiponectin, and fatty acid binding protein (FABP)4 involved in adipogenesis was determined by western blot analysis. TJF extract effectively inhibited lipid accumulation and the expression of PPARγ and C/EBPα in 3T3-L1 cells. TJF also increased the phosphorylation of AMPK and ACC, and decreased the expression of adiponectin and FABP4. These results indicate that TJF extract exerts its anti-obesity effect through the downregulation of adipogenic transcription factors and adipogenic marker genes.展开更多
The peripheral nervous system has the potential to regenerate after nerve injury owing to the intrinsic regrowth ability of neurons and the permissive microenvironment.The regenerative process involves numerous gene e...The peripheral nervous system has the potential to regenerate after nerve injury owing to the intrinsic regrowth ability of neurons and the permissive microenvironment.The regenerative process involves numerous gene expression changes,in which transcription factors play a critical role.Previously,we profiled dysregulated genes in dorsal root ganglion neurons at different time points（0,3 and 9 hours,and 1,4 and 7 days） after sciatic nerve injury in rats by RNA sequencing.In the present study,we investigated differentially expressed transcription factors following nerve injury,and we identified enriched molecular and cellular functions of these transcription factors by Ingenuity Pathway Analysis.This analysis revealed the dynamic changes in the expression of transcription factors involved in cell death at different time points following sciatic nerve injury.In addition,we constructed regulatory networks of the differentially expressed transcription factors in cell death and identified some key transcription factors（such as STAT1,JUN,MYC and IRF7）.We confirmed the changes in expression of some key transcription factors（STAT1 and IRF7） by quantitative reverse transcription-polymerase chain reaction.Collectively,our analyses provide a global overview of transcription factor changes in dorsal root ganglia after sciatic nerve injury and offer insight into the regulatory transcription factor networks involved in cell death.展开更多
Poly(ADP-ribose) polymerase 1 (PARP1) plays important roles in the regulation of transcription factors. Mounting evidence has shown that inhibition of PARP1 influences the expression of genes associated with inflammat...Poly(ADP-ribose) polymerase 1 (PARP1) plays important roles in the regulation of transcription factors. Mounting evidence has shown that inhibition of PARP1 influences the expression of genes associated with inflammatory response. Interferon regulatory factor 1 (IRF1) is a critical transcription factor for the development of both the innate and adaptive immune responses against infections. However, the molecular mechanism through which PARP1 mediates the effects has not been clearly demonstrated. Jurkat cells were exposed to dexamethasone (Dex) or PARP1 inhibitor PJ34. The expression levels of IL-12, LMP2, OAS1 and PKR were detected using real-time RT-PCR. The interactions between PARP1 and IRF1 were examined by coimmunoprecipitation (co-IP) assays. We further explored the mechanism of PARP1 suppressing IRF1 by assessing the activities of interferon stimulated response element (ISRE). The mRNA expression of IL-12, LMP2, OAS1 and PKR was obviously suppressed by Dex in Jurkat cells, which could be rescued by PJ34 treatment. Luciferase study revealed that poly(ADP-ribosyl)- ation suppressed IRF1-mediated transcription through preventing the binding of IRF1 to ISREs. PARP1 inhibited IRF1-mediated transcription in Jurkat cells by preventing IRF1 binding to ISREs in the promoters of target genes. It is suggested that PARP1 is a crucial regulator of IRF1-mediated immune response. This study provides experimental evidence for the possible application of PARP1 inhibitors in the treatment of IRF1-related immune anergy.展开更多
Expression of P-selectin in injured or activated endothelia cells serves as a permissive step towards leukocyte recruitment and perpetuation of inflammation in the pathogenesis of atherosclerosis.P-selectin can be ind...Expression of P-selectin in injured or activated endothelia cells serves as a permissive step towards leukocyte recruitment and perpetuation of inflammation in the pathogenesis of atherosclerosis.P-selectin can be induced by pro-inflammatory stimuli via the transcription factor NF-κB,but the epigenetic mechanisms remain incompletely understood.Previously we reported that myocardin-related transcription factor A（MRTF-A）mediates the transactivation of a slew of adhesion molecules by oxidized low-density lipoprotein（oxLDL）,likely through a crosstalk with brahma-related gene 1（BRGl）,a chromatin remodeling protein.Here,we show that MRTF-A was both sufficient and necessary for the transactivation of P-selectin gene in endothelial cells treated with TNF-α.Depletion of MRTF-A using small interfering RNA（siRNA）abrogated the binding of BRGl on the P-selectin promoter.Overexpression of BRG1 up-regulated the activity of P-selectin promoter activity while BRGl knockdown attenuated P-selectin expression.Finally,BRGl silencing suppressed the accumulation of acetylated histone H3 and methylated histone H3K4,and altered the binding of NF-κB on the P-selectin promoter.Therefore,our data demonstrate an essential role for MRTF-A and BRGl in P-selectin transactivation in endothelial cells.展开更多
AIM To observe the growth suppression effect of exogenous introduction of early growth response gene-1 (Egr-1 gene) on esophageal carcinoma tissue as well as on esophageal carcinoma cell line Eca109 and to explore the...AIM To observe the growth suppression effect of exogenous introduction of early growth response gene-1 (Egr-1 gene) on esophageal carcinoma tissue as well as on esophageal carcinoma cell line Eca109 and to explore the potential application of Egr-1 gene in gene therapy of tumor.METHODS Eukaryotic expression vector of PCMV-Egr-1 plasmid was introduced into Eca109 cell line which expressed no Egr-1 protein originally with lipofectamine transfection method. The introduction and expression of PCMV-Egr-1 plasmid into Eca109 cell line was confirmed by G418 selection culture, PCR amplification of neogene contained in the vector, Western blot analysis and immunocytochemical analysis. The cell growth curve,soft agar colony formation rate and tumorigenicity in SClD mice were examined to demonstrate the growth suppression effect of exogenous Egr-1 gene on Eca109 cell line. The Egr-1 mRNA and Egr-1 protein were also detected in 50 surgical specimens of esophageal carcinoma by in situ hybridization and immunohistochemistry.RESULTS Exogenous Egr-1 gene was introduced successfully into Eca109 cell line and expressed Egr-1 protein stably. The transfected Eca109 cell line grew more slowly than control Eca109 as shown by cell growth curves, the soft agar colony formation rate (4.0% vs 6.9%, P＜0.01) and the average growth rate of tumor in SCID mice (35.5 ± 7.6 vs 65.8 ± 7.6, P＜0.05). The expression level of Egr-1 mRNA and protein significantly increased in dysplastic epithelia adjacent to cancer rather than in cancer tissues (65.8% vs 20.0% by ISH and 57.9% vs 14.0% by IHC, P＜0.01).CONCLUSION Exogenous Egr-1 gene shows the strong effect of growth inhibition in Eca109 cell line. Egr-1 in the cancer tissue shows down-regulated expression that supports the inhibited function of Egr-1 in cancer growth and suggests Egr-1 may have an important role in gene therapy of esophageal carcinoma.展开更多
目的探讨胃癌组织中核转录因子-κB(nuclear factor kappa B,NF-κB)、程序性细胞死亡蛋白-1(programmed cell death protein-1,PD-1)、程序性细胞死亡蛋白配体-1(programmed cell death protein ligand 1,PD-L1)的表达及其临床意义。...目的探讨胃癌组织中核转录因子-κB(nuclear factor kappa B,NF-κB)、程序性细胞死亡蛋白-1(programmed cell death protein-1,PD-1)、程序性细胞死亡蛋白配体-1(programmed cell death protein ligand 1,PD-L1)的表达及其临床意义。方法选择2021年1月至2023年12月于南阳市第一人民医院治疗的62例胃癌患者手术切除后组织标本及癌旁正常组织标本分别作为胃癌组及癌旁组。62例患者中,男39例,女23例,年龄(61.86±2.31)岁,肿瘤长径(4.92±0.53)cm。采用免疫组织化学染色法检测NF-κB、PD-1、PD-L1表达情况。比较两组NF-κB、PD-1、PD-L1表达阳性率,分析其与胃癌临床病理特征的关系。采用χ^(2)检验。结果胃癌组NF-κB、PD-1、PD-L1表达阳性率高于癌旁组[83.87%(52/62)比33.87%(21/62)、75.81%(47/62)比30.65%(19/62)、80.65%(50/62)比40.32%(25/62)],差异均有统计学意义(χ^(2)=32.008、25.396、21.088,均P<0.05);肌层及浆膜层浸润、有淋巴结转移及Ⅲ+Ⅳ期患者NF-κB表达阳性率高于黏膜及黏膜下层浸润、无淋巴结转移及Ⅰ+Ⅱ期患者[92.68%(38/41)比66.67%(14/21)、94.74(36/38)比66.67%(16/24)、92.31%(36/39)比69.57%(16/23)],差异均有统计学意义(χ^(2)=5.158、6.619、3.978,均P<0.05);肌层及浆膜层浸润、有淋巴结转移及Ⅲ+Ⅳ期患者PD-1表达阳性率高于黏膜及黏膜下层浸润、无淋巴结转移及Ⅰ+Ⅱ期患者[85.37%(35/41)比57.14%(14/21)、89.47%(34/38)比54.17%(13/24)、89.74%(35/39)比52.17%(12/23)],差异均有统计学意义(χ^(2)=6.031、9.998、11.135,均P<0.05);低分化、肌层及浆膜层浸润、有淋巴结转移及Ⅲ+Ⅳ期患者PD-L1表达阳性率高于高中分化、黏膜及黏膜下层浸润、无淋巴结转移及Ⅰ+Ⅱ期患者[96.00%(24/25)比70.27%(26/37)、90.24%(37/41)比61.90%(13/21)、92.11%(35/38)比62.50%(15/24)、94.87%(37/39)比56.52%(13/23)],差异均有统计学意义(χ^(2)=4.787、5.445、6.472、11.286,均P<0.05)。结论胃癌组织中NF-κB、PD-1、PD-L1表达阳性率升高,且NF-κB、PD-1表达与胃癌浸润深度、淋巴结转移及TNM分期有关,PD-L1表达与胃癌分化程度、浸润深度、淋巴结转移及TNM分期有关。展开更多
文摘Obesity is a major human health problem associated with various diseases, including cardiac injury and type 2 diabetes. Trapa japonica Flerov (TJF) has been used in traditional oriental medicine to treat diabetes. In this study, we evaluated the inhibitory effect of and the mechanism underlying the effect of TJF extract on adipogenesis in 3T3-L1 cells. The effects of TJF extract on cell viability were analyzed using a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay, and the anti-adipogenic effect was measured by oil red O staining. The expression of peroxisomal proliferator activated receptor (PPAR)γ, CCAAT/enhancer-binding protein-α (C/EBP)α, adenosine monophosphate-activated protein kinase (AMPK), acetyl-CoA carboxylase (ACC), adiponectin, and fatty acid binding protein (FABP)4 involved in adipogenesis was determined by western blot analysis. TJF extract effectively inhibited lipid accumulation and the expression of PPARγ and C/EBPα in 3T3-L1 cells. TJF also increased the phosphorylation of AMPK and ACC, and decreased the expression of adiponectin and FABP4. These results indicate that TJF extract exerts its anti-obesity effect through the downregulation of adipogenic transcription factors and adipogenic marker genes.
基金supported by the National Natural Science Foundation of China,No.31500823the Natural Science Foundation of Jiangsu Province of China,No.BK20150403the Natural Science Fund for Colleges and Universities in Jiangsu Province of China,No.16KJB180024
文摘The peripheral nervous system has the potential to regenerate after nerve injury owing to the intrinsic regrowth ability of neurons and the permissive microenvironment.The regenerative process involves numerous gene expression changes,in which transcription factors play a critical role.Previously,we profiled dysregulated genes in dorsal root ganglion neurons at different time points（0,3 and 9 hours,and 1,4 and 7 days） after sciatic nerve injury in rats by RNA sequencing.In the present study,we investigated differentially expressed transcription factors following nerve injury,and we identified enriched molecular and cellular functions of these transcription factors by Ingenuity Pathway Analysis.This analysis revealed the dynamic changes in the expression of transcription factors involved in cell death at different time points following sciatic nerve injury.In addition,we constructed regulatory networks of the differentially expressed transcription factors in cell death and identified some key transcription factors（such as STAT1,JUN,MYC and IRF7）.We confirmed the changes in expression of some key transcription factors（STAT1 and IRF7） by quantitative reverse transcription-polymerase chain reaction.Collectively,our analyses provide a global overview of transcription factor changes in dorsal root ganglia after sciatic nerve injury and offer insight into the regulatory transcription factor networks involved in cell death.
基金This work was supported by the National Natural Science Foundation of China (No.81370263 and No.81500348).
文摘Poly(ADP-ribose) polymerase 1 (PARP1) plays important roles in the regulation of transcription factors. Mounting evidence has shown that inhibition of PARP1 influences the expression of genes associated with inflammatory response. Interferon regulatory factor 1 (IRF1) is a critical transcription factor for the development of both the innate and adaptive immune responses against infections. However, the molecular mechanism through which PARP1 mediates the effects has not been clearly demonstrated. Jurkat cells were exposed to dexamethasone (Dex) or PARP1 inhibitor PJ34. The expression levels of IL-12, LMP2, OAS1 and PKR were detected using real-time RT-PCR. The interactions between PARP1 and IRF1 were examined by coimmunoprecipitation (co-IP) assays. We further explored the mechanism of PARP1 suppressing IRF1 by assessing the activities of interferon stimulated response element (ISRE). The mRNA expression of IL-12, LMP2, OAS1 and PKR was obviously suppressed by Dex in Jurkat cells, which could be rescued by PJ34 treatment. Luciferase study revealed that poly(ADP-ribosyl)- ation suppressed IRF1-mediated transcription through preventing the binding of IRF1 to ISREs. PARP1 inhibited IRF1-mediated transcription in Jurkat cells by preventing IRF1 binding to ISREs in the promoters of target genes. It is suggested that PARP1 is a crucial regulator of IRF1-mediated immune response. This study provides experimental evidence for the possible application of PARP1 inhibitors in the treatment of IRF1-related immune anergy.
基金supported by the Natural Science Foundation of Jiangsu Province(BK20141498)a grant from Jiangsu Jiankang Vocational University(JKC201505)
文摘Expression of P-selectin in injured or activated endothelia cells serves as a permissive step towards leukocyte recruitment and perpetuation of inflammation in the pathogenesis of atherosclerosis.P-selectin can be induced by pro-inflammatory stimuli via the transcription factor NF-κB,but the epigenetic mechanisms remain incompletely understood.Previously we reported that myocardin-related transcription factor A（MRTF-A）mediates the transactivation of a slew of adhesion molecules by oxidized low-density lipoprotein（oxLDL）,likely through a crosstalk with brahma-related gene 1（BRGl）,a chromatin remodeling protein.Here,we show that MRTF-A was both sufficient and necessary for the transactivation of P-selectin gene in endothelial cells treated with TNF-α.Depletion of MRTF-A using small interfering RNA（siRNA）abrogated the binding of BRGl on the P-selectin promoter.Overexpression of BRG1 up-regulated the activity of P-selectin promoter activity while BRGl knockdown attenuated P-selectin expression.Finally,BRGl silencing suppressed the accumulation of acetylated histone H3 and methylated histone H3K4,and altered the binding of NF-κB on the P-selectin promoter.Therefore,our data demonstrate an essential role for MRTF-A and BRGl in P-selectin transactivation in endothelial cells.
基金Supported by the National Natural Science Foundation of China,No.39670298.
文摘AIM To observe the growth suppression effect of exogenous introduction of early growth response gene-1 (Egr-1 gene) on esophageal carcinoma tissue as well as on esophageal carcinoma cell line Eca109 and to explore the potential application of Egr-1 gene in gene therapy of tumor.METHODS Eukaryotic expression vector of PCMV-Egr-1 plasmid was introduced into Eca109 cell line which expressed no Egr-1 protein originally with lipofectamine transfection method. The introduction and expression of PCMV-Egr-1 plasmid into Eca109 cell line was confirmed by G418 selection culture, PCR amplification of neogene contained in the vector, Western blot analysis and immunocytochemical analysis. The cell growth curve,soft agar colony formation rate and tumorigenicity in SClD mice were examined to demonstrate the growth suppression effect of exogenous Egr-1 gene on Eca109 cell line. The Egr-1 mRNA and Egr-1 protein were also detected in 50 surgical specimens of esophageal carcinoma by in situ hybridization and immunohistochemistry.RESULTS Exogenous Egr-1 gene was introduced successfully into Eca109 cell line and expressed Egr-1 protein stably. The transfected Eca109 cell line grew more slowly than control Eca109 as shown by cell growth curves, the soft agar colony formation rate (4.0% vs 6.9%, P＜0.01) and the average growth rate of tumor in SCID mice (35.5 ± 7.6 vs 65.8 ± 7.6, P＜0.05). The expression level of Egr-1 mRNA and protein significantly increased in dysplastic epithelia adjacent to cancer rather than in cancer tissues (65.8% vs 20.0% by ISH and 57.9% vs 14.0% by IHC, P＜0.01).CONCLUSION Exogenous Egr-1 gene shows the strong effect of growth inhibition in Eca109 cell line. Egr-1 in the cancer tissue shows down-regulated expression that supports the inhibited function of Egr-1 in cancer growth and suggests Egr-1 may have an important role in gene therapy of esophageal carcinoma.
文摘目的探讨胃癌组织中核转录因子-κB(nuclear factor kappa B,NF-κB)、程序性细胞死亡蛋白-1(programmed cell death protein-1,PD-1)、程序性细胞死亡蛋白配体-1(programmed cell death protein ligand 1,PD-L1)的表达及其临床意义。方法选择2021年1月至2023年12月于南阳市第一人民医院治疗的62例胃癌患者手术切除后组织标本及癌旁正常组织标本分别作为胃癌组及癌旁组。62例患者中,男39例,女23例,年龄(61.86±2.31)岁,肿瘤长径(4.92±0.53)cm。采用免疫组织化学染色法检测NF-κB、PD-1、PD-L1表达情况。比较两组NF-κB、PD-1、PD-L1表达阳性率,分析其与胃癌临床病理特征的关系。采用χ^(2)检验。结果胃癌组NF-κB、PD-1、PD-L1表达阳性率高于癌旁组[83.87%(52/62)比33.87%(21/62)、75.81%(47/62)比30.65%(19/62)、80.65%(50/62)比40.32%(25/62)],差异均有统计学意义(χ^(2)=32.008、25.396、21.088,均P<0.05);肌层及浆膜层浸润、有淋巴结转移及Ⅲ+Ⅳ期患者NF-κB表达阳性率高于黏膜及黏膜下层浸润、无淋巴结转移及Ⅰ+Ⅱ期患者[92.68%(38/41)比66.67%(14/21)、94.74(36/38)比66.67%(16/24)、92.31%(36/39)比69.57%(16/23)],差异均有统计学意义(χ^(2)=5.158、6.619、3.978,均P<0.05);肌层及浆膜层浸润、有淋巴结转移及Ⅲ+Ⅳ期患者PD-1表达阳性率高于黏膜及黏膜下层浸润、无淋巴结转移及Ⅰ+Ⅱ期患者[85.37%(35/41)比57.14%(14/21)、89.47%(34/38)比54.17%(13/24)、89.74%(35/39)比52.17%(12/23)],差异均有统计学意义(χ^(2)=6.031、9.998、11.135,均P<0.05);低分化、肌层及浆膜层浸润、有淋巴结转移及Ⅲ+Ⅳ期患者PD-L1表达阳性率高于高中分化、黏膜及黏膜下层浸润、无淋巴结转移及Ⅰ+Ⅱ期患者[96.00%(24/25)比70.27%(26/37)、90.24%(37/41)比61.90%(13/21)、92.11%(35/38)比62.50%(15/24)、94.87%(37/39)比56.52%(13/23)],差异均有统计学意义(χ^(2)=4.787、5.445、6.472、11.286,均P<0.05)。结论胃癌组织中NF-κB、PD-1、PD-L1表达阳性率升高,且NF-κB、PD-1表达与胃癌浸润深度、淋巴结转移及TNM分期有关,PD-L1表达与胃癌分化程度、浸润深度、淋巴结转移及TNM分期有关。
