In order to clarify the mechanism of action of licorice flavonoids in alleviating bone loss caused by osteoporosis,this study compared the effects of four glycyrrhiza flavonoids,naringenin,liquiritigenin,isoliquiritig...In order to clarify the mechanism of action of licorice flavonoids in alleviating bone loss caused by osteoporosis,this study compared the effects of four glycyrrhiza flavonoids,naringenin,liquiritigenin,isoliquiritigenin,and licochalcone A,on osteogenic differentiation and mineralization by molecular docking simulation,alkaline phosphatase(ALP)activity and osteocalcin(OCN)content assays,and Runt-related transcription factor 2(Runx2)expression,and explored their potential molecular mechanisms.The results of molecular docking showed that the docking score of liquiritigenin with the estrogen receptor(ER)was the highest.All four flavonoids up-regulated ALP activity and OCN concentration in MC3T3-E1 cells,thereby elevating the mineralization level,among which liquiritigenin was the most effective.Moreover,treatment with a phosphatidylinositol-3-kinase(PI3K)inhibitor(LY294002)inhibited liquiritigenin from inducing increased phosphorylation levels in the PI3K/protein kinase B(AKT)signaling pathway and up-regulation of Runx2 expression,suggesting that PI3K and AKT were involved in osteogenic action.Liquiritigenin reversed bone mineral density loss in a zebrafish osteoporosis model.These findings suggest that liquiritigenin has the most significant osteogenic effect among the four estrogen-like flavonoids,stimulating osteoblast differentiation and bone mineralization through the activation of Runx2 via the PI3K/AKT signaling pathways.In conclusion,this study highlights the great potential of liquiritigenin for preventing and treating osteoporosis.展开更多
The trace element selenium(Se)occurs naturally throughout the earth.Se deficiency has been linked to impaired breast health and other diseases in human and animals.Compared to severe Se deficiency,marginal dietary Se ...The trace element selenium(Se)occurs naturally throughout the earth.Se deficiency has been linked to impaired breast health and other diseases in human and animals.Compared to severe Se deficiency,marginal dietary Se deficiency accusers more frequently in low-Se regions.Therefore,to investigate the Se status and inflammatory response of the mammary gland under marginal dietary Se levels,an lipopolysaccharide(LPS)induced mouse mastitis model was established.Mice were fed with moderate Se diet(0.087 mg•kg^(-1) Se),adequate Se diet(0.15 mg•kg^(-1) Se)or excessive Se diet(1.5 mg•kg^(-1) Se)for 60 days.Se status and inflammatory factors were investigated.Results showed that the Se status of mammary gland correlated with dietary Se levels.Marginal Se deficiency exacerbated mammary tissue histopathology;increased the mRNA level of inflammatory genes tumor necrosis factor alpha(TNF-α),interleukin-1β(IL-1β)and cyclooxygenase-2(COX-2);and enhanced the phosphorylation of NF-κB p65 in mammary gland tissues.Supplementation of Se in diet higher than recommended levels reduced the inflammatory reaction of mammary glands in LPS-induced mastitis model and provided a protective effect.展开更多
Objective To investigate the effect of mucin 1(MUC1)on the proliferation and apoptosis of nasopharyngeal carcinoma(NPC)and its regulatory mechanism.Methods The 60 NPC and paired para-cancer normal tissues were collect...Objective To investigate the effect of mucin 1(MUC1)on the proliferation and apoptosis of nasopharyngeal carcinoma(NPC)and its regulatory mechanism.Methods The 60 NPC and paired para-cancer normal tissues were collected from October 2020 to July 2021 in Quanzhou First Hospital.The expression of MUC1 was measured by real-time quantitative PCR(qPCR)in the patients with PNC.The 5-8F and HNE1 cells were transfected with siRNA control(si-control)or siRNA targeting MUC1(si-MUC1).Cell proliferation was analyzed by cell counting kit-8 and colony formation assay,and apoptosis was analyzed by flow cytometry analysis in the 5-8F and HNE1 cells.The qPCR and ELISA were executed to analyze the levels of TNF-αand IL-6.Western blot was performed to measure the expression of MUC1,NFкB and apoptosis-related proteins(Bax and Bcl-2).Results The expression of MUC1 was up-regulated in the NPC tissues,and NPC patients with the high MUC1 expression were inclined to EBV infection,growth and metastasis of NPC.Loss of MUC1 restrained malignant features,including the proliferation and apoptosis,downregulated the expression of p-IкB、p-P65 and Bcl-2 and upregulated the expression of Bax in the NPC cells.Conclusion Downregulation of MUC1 restrained biological characteristics of malignancy,including cell proliferation and apoptosis,by inactivating NF-κB signaling pathway in NPC.展开更多
目的基于核因子激活的B细胞的NOD-样受体热蛋白结构域相关蛋白3(NOD-like receptor pyrin domain-containing protein 3,NLRP3)/白细胞介素-18(interleukin-18,IL-18)/κ-轻链(nuclear factor-activated B cellκ-light chain,NF-κB)...目的基于核因子激活的B细胞的NOD-样受体热蛋白结构域相关蛋白3(NOD-like receptor pyrin domain-containing protein 3,NLRP3)/白细胞介素-18(interleukin-18,IL-18)/κ-轻链(nuclear factor-activated B cellκ-light chain,NF-κB)信号通路探讨化痰安神汤抑制神经元细胞凋亡的作用机制。方法体外培养HT22小鼠海马神经元细胞,C57小鼠灌胃化痰安神方后制备含药小鼠血清,CCK-8法筛选化痰安神方含药血清最佳干预浓度。采用不同浓度含药血清处理HT22细胞后,予脂多糖(lipopolysaccharide,LPS)诱导其发生凋亡,CCK-8法L-乳酸脱氢酶(L-lactate dehydrogenase,L-LDH)生化检测法和Annexin V/PI染色法用于检测细胞损伤与凋亡情况,Western blotting法用于测定通路相关蛋白P-NF-κB/NF-κB、凋亡相关的斑点样蛋白(apoptosis-associated speck-like protein containing,ASC)、NLRP3、Caspase-1的表达,酶联免疫吸附测定法用于测定IL-18的表达。过表达NLRP3后重复试验,进行单基因功能验证。结果化痰安神方含药血清可改善LPS诱导的细胞损伤和凋亡,降低细胞L-LDH浓度(P<0.05);显著下调NLRP3/IL-18/NF-κB信号通路相关蛋白的转录表达(P<0.05);显著降低细胞内IL-18浓度;同时过表达NLRP3可逆转化痰安神方含药血清对HT22细胞凋亡抑制作用(P<0.05)。结论化痰安神方能够通过抑制NLRP3/IL-18/NF-κB信号通路改善神经元细胞凋亡。展开更多
为探究羊口疮病毒(ORFV)129蛋白对NF-κB信号通路介导的宿主细胞因子转录水平的影响,本研究首先设计并合成山羊4种细胞因子基因特异性引物,分别以构建的4种细胞因子重组质粒为模板,经条件优化建立了检测山羊4种细胞因子的SYBR Green I...为探究羊口疮病毒(ORFV)129蛋白对NF-κB信号通路介导的宿主细胞因子转录水平的影响,本研究首先设计并合成山羊4种细胞因子基因特异性引物,分别以构建的4种细胞因子重组质粒为模板,经条件优化建立了检测山羊4种细胞因子的SYBR Green I荧光定量RT-PCR方法。结果显示,所建立的4种细胞因子SYBR Green I荧光定量RT-PCR方法熔解曲线呈单一峰,无引物二聚体;4种细胞因子重组质粒标准品最低检测限分别为IL-6:1.40×10^(2)拷贝/μL、IL-1β:1.92×10^(2)拷贝/μL、IFN-γ:1.69×10^(2)拷贝/μL、TNF-α:2.31×10^(1)拷贝/μL;组内及组间变异系数在0.1%~2.3%。表明所建立的山羊4种细胞因子荧光定量RT-PCR特异性强,灵敏度高,稳定性好,可用于4种细胞因子mRNA转录水平的检测。随后本研究将包含编码ORFV 129完整蛋白基因的重组质粒pEGFP-ORFV 129和pEGFP-N1空质粒分别转染山羊睾丸支持细胞(GTSCs)24 h后,用终浓度20 ng/mL的TNF-α刺激转染细胞6 h后收集细胞,利用建立的荧光定量RT-PCR方法检测GTSCs中IL-1β、IL-6、IFN-γ和TNF-α共4种细胞因子mRNA的转录水平,结果显示,与转染pEGFP-N1空载的细胞相比,转染了pEGFP-ORFV 129重组质粒的细胞IL-1β、IL-6和IFN-γmRNA的转录水平均极显著下降(p<0.001),该细胞中TNF-α的mRNA转录水平也下降,但与对照组相比差异不显著(p>0.1),表明ORFV 129蛋白能够抑制GTSCs中NF-κB信号通路介导的IL-1β、IL-6、IFN-γ和TNF-αmRNA的转录水平。本研究通过建立山羊IL-1β、IL-6、IFN-γ和TNF-α4种细胞因子的SYBR Green I荧光定量PCR方法,首次检测证实了ORFV 129蛋白不同程度地抑制GTSCs中NF-κB信号通路介导的IL-1β、IL-6、IFN-γ和TNF-αmRNA的转录水平,为ORFV129蛋白功能研究奠定了基础。展开更多
Transcription factor NF-κB has attracted attention due to its important role in the regulation of the expression of a number of cellular genes involved in host defense,inflammation/immune responses,cytokine and growt...Transcription factor NF-κB has attracted attention due to its important role in the regulation of the expression of a number of cellular genes involved in host defense,inflammation/immune responses,cytokine and growth factor regulation,cell survival,proliferation,and also in embryonic development and programmed cell death. In this review,we provide an overview of established TNF-α,TLR/IL-1R,TCR and BCR signaling pathways to NF-κB,the alternative NF-κB pathway,and the regulation of NF-κB transcriptional activity. Furthermore,we discuss on emphasis the cross-talks between NF-κB and other signaling pathways (TGF-β,p53,nuclear receptor family,Ras/MEK/MAPK...),which are very complex and remain to be understood. The activation or inhibition of NF-κB has effect on other signaling pathways. They compose a complex signaling network,in which many regulatory signals integrate systematically and coordinate transcriptional responses to the stimulations. So when NF-κB is used as an important target of drugs in human diseases,the pathological and physiological functions of related signal pathways also should be understood sufficiently.展开更多
IκB is an inhibitor of nuclear factor-kappa B(NF-κB) and presents in the majority of cells.Eightstructurally related members of the mammalian IκB family have been described: IκBα,IκBβ,IκBε,Bcl-3,IκBγ,IκBδ...IκB is an inhibitor of nuclear factor-kappa B(NF-κB) and presents in the majority of cells.Eightstructurally related members of the mammalian IκB family have been described: IκBα,IκBβ,IκBε,Bcl-3,IκBγ,IκBδ,p100 and p105.The ankyrin repeat domain of IκB can interact with NF-κB,and sequester NF-κB in the cytoplasm as in-active complexes.Most recently,IκBαhas been found to inhibit p53 tumor suppressor protein by binding p53 to form a cyto-plasmic p53.IκBαcomplex and studies have shown that p100 profoundly sensitizes cells to death-receptor-mediated ap-optosis.The current review is to describe the members of IκB family,their related signaling pathways,and their applicationin tumor therapy.展开更多
文摘In order to clarify the mechanism of action of licorice flavonoids in alleviating bone loss caused by osteoporosis,this study compared the effects of four glycyrrhiza flavonoids,naringenin,liquiritigenin,isoliquiritigenin,and licochalcone A,on osteogenic differentiation and mineralization by molecular docking simulation,alkaline phosphatase(ALP)activity and osteocalcin(OCN)content assays,and Runt-related transcription factor 2(Runx2)expression,and explored their potential molecular mechanisms.The results of molecular docking showed that the docking score of liquiritigenin with the estrogen receptor(ER)was the highest.All four flavonoids up-regulated ALP activity and OCN concentration in MC3T3-E1 cells,thereby elevating the mineralization level,among which liquiritigenin was the most effective.Moreover,treatment with a phosphatidylinositol-3-kinase(PI3K)inhibitor(LY294002)inhibited liquiritigenin from inducing increased phosphorylation levels in the PI3K/protein kinase B(AKT)signaling pathway and up-regulation of Runx2 expression,suggesting that PI3K and AKT were involved in osteogenic action.Liquiritigenin reversed bone mineral density loss in a zebrafish osteoporosis model.These findings suggest that liquiritigenin has the most significant osteogenic effect among the four estrogen-like flavonoids,stimulating osteoblast differentiation and bone mineralization through the activation of Runx2 via the PI3K/AKT signaling pathways.In conclusion,this study highlights the great potential of liquiritigenin for preventing and treating osteoporosis.
基金Supported by the Natural Science Foundation of the Heilongjiang Province of China(C2016003)China Postdoctoral Science Foundation(2015M581415)Heilongjiang Postdoctoral Fund(LBH-Z15005)。
文摘The trace element selenium(Se)occurs naturally throughout the earth.Se deficiency has been linked to impaired breast health and other diseases in human and animals.Compared to severe Se deficiency,marginal dietary Se deficiency accusers more frequently in low-Se regions.Therefore,to investigate the Se status and inflammatory response of the mammary gland under marginal dietary Se levels,an lipopolysaccharide(LPS)induced mouse mastitis model was established.Mice were fed with moderate Se diet(0.087 mg•kg^(-1) Se),adequate Se diet(0.15 mg•kg^(-1) Se)or excessive Se diet(1.5 mg•kg^(-1) Se)for 60 days.Se status and inflammatory factors were investigated.Results showed that the Se status of mammary gland correlated with dietary Se levels.Marginal Se deficiency exacerbated mammary tissue histopathology;increased the mRNA level of inflammatory genes tumor necrosis factor alpha(TNF-α),interleukin-1β(IL-1β)and cyclooxygenase-2(COX-2);and enhanced the phosphorylation of NF-κB p65 in mammary gland tissues.Supplementation of Se in diet higher than recommended levels reduced the inflammatory reaction of mammary glands in LPS-induced mastitis model and provided a protective effect.
文摘Objective To investigate the effect of mucin 1(MUC1)on the proliferation and apoptosis of nasopharyngeal carcinoma(NPC)and its regulatory mechanism.Methods The 60 NPC and paired para-cancer normal tissues were collected from October 2020 to July 2021 in Quanzhou First Hospital.The expression of MUC1 was measured by real-time quantitative PCR(qPCR)in the patients with PNC.The 5-8F and HNE1 cells were transfected with siRNA control(si-control)or siRNA targeting MUC1(si-MUC1).Cell proliferation was analyzed by cell counting kit-8 and colony formation assay,and apoptosis was analyzed by flow cytometry analysis in the 5-8F and HNE1 cells.The qPCR and ELISA were executed to analyze the levels of TNF-αand IL-6.Western blot was performed to measure the expression of MUC1,NFкB and apoptosis-related proteins(Bax and Bcl-2).Results The expression of MUC1 was up-regulated in the NPC tissues,and NPC patients with the high MUC1 expression were inclined to EBV infection,growth and metastasis of NPC.Loss of MUC1 restrained malignant features,including the proliferation and apoptosis,downregulated the expression of p-IкB、p-P65 and Bcl-2 and upregulated the expression of Bax in the NPC cells.Conclusion Downregulation of MUC1 restrained biological characteristics of malignancy,including cell proliferation and apoptosis,by inactivating NF-κB signaling pathway in NPC.
文摘为探究羊口疮病毒(ORFV)129蛋白对NF-κB信号通路介导的宿主细胞因子转录水平的影响,本研究首先设计并合成山羊4种细胞因子基因特异性引物,分别以构建的4种细胞因子重组质粒为模板,经条件优化建立了检测山羊4种细胞因子的SYBR Green I荧光定量RT-PCR方法。结果显示,所建立的4种细胞因子SYBR Green I荧光定量RT-PCR方法熔解曲线呈单一峰,无引物二聚体;4种细胞因子重组质粒标准品最低检测限分别为IL-6:1.40×10^(2)拷贝/μL、IL-1β:1.92×10^(2)拷贝/μL、IFN-γ:1.69×10^(2)拷贝/μL、TNF-α:2.31×10^(1)拷贝/μL;组内及组间变异系数在0.1%~2.3%。表明所建立的山羊4种细胞因子荧光定量RT-PCR特异性强,灵敏度高,稳定性好,可用于4种细胞因子mRNA转录水平的检测。随后本研究将包含编码ORFV 129完整蛋白基因的重组质粒pEGFP-ORFV 129和pEGFP-N1空质粒分别转染山羊睾丸支持细胞(GTSCs)24 h后,用终浓度20 ng/mL的TNF-α刺激转染细胞6 h后收集细胞,利用建立的荧光定量RT-PCR方法检测GTSCs中IL-1β、IL-6、IFN-γ和TNF-α共4种细胞因子mRNA的转录水平,结果显示,与转染pEGFP-N1空载的细胞相比,转染了pEGFP-ORFV 129重组质粒的细胞IL-1β、IL-6和IFN-γmRNA的转录水平均极显著下降(p<0.001),该细胞中TNF-α的mRNA转录水平也下降,但与对照组相比差异不显著(p>0.1),表明ORFV 129蛋白能够抑制GTSCs中NF-κB信号通路介导的IL-1β、IL-6、IFN-γ和TNF-αmRNA的转录水平。本研究通过建立山羊IL-1β、IL-6、IFN-γ和TNF-α4种细胞因子的SYBR Green I荧光定量PCR方法,首次检测证实了ORFV 129蛋白不同程度地抑制GTSCs中NF-κB信号通路介导的IL-1β、IL-6、IFN-γ和TNF-αmRNA的转录水平,为ORFV129蛋白功能研究奠定了基础。
文摘Transcription factor NF-κB has attracted attention due to its important role in the regulation of the expression of a number of cellular genes involved in host defense,inflammation/immune responses,cytokine and growth factor regulation,cell survival,proliferation,and also in embryonic development and programmed cell death. In this review,we provide an overview of established TNF-α,TLR/IL-1R,TCR and BCR signaling pathways to NF-κB,the alternative NF-κB pathway,and the regulation of NF-κB transcriptional activity. Furthermore,we discuss on emphasis the cross-talks between NF-κB and other signaling pathways (TGF-β,p53,nuclear receptor family,Ras/MEK/MAPK...),which are very complex and remain to be understood. The activation or inhibition of NF-κB has effect on other signaling pathways. They compose a complex signaling network,in which many regulatory signals integrate systematically and coordinate transcriptional responses to the stimulations. So when NF-κB is used as an important target of drugs in human diseases,the pathological and physiological functions of related signal pathways also should be understood sufficiently.
文摘IκB is an inhibitor of nuclear factor-kappa B(NF-κB) and presents in the majority of cells.Eightstructurally related members of the mammalian IκB family have been described: IκBα,IκBβ,IκBε,Bcl-3,IκBγ,IκBδ,p100 and p105.The ankyrin repeat domain of IκB can interact with NF-κB,and sequester NF-κB in the cytoplasm as in-active complexes.Most recently,IκBαhas been found to inhibit p53 tumor suppressor protein by binding p53 to form a cyto-plasmic p53.IκBαcomplex and studies have shown that p100 profoundly sensitizes cells to death-receptor-mediated ap-optosis.The current review is to describe the members of IκB family,their related signaling pathways,and their applicationin tumor therapy.
