Loss-of-function variants of low-density lipoprotein receptor-related protein 5(LRP5)can lead to reduced bone formation,culminating in diminished bone mass.Our previous study reported transcription factor osterix(SP7)...Loss-of-function variants of low-density lipoprotein receptor-related protein 5(LRP5)can lead to reduced bone formation,culminating in diminished bone mass.Our previous study reported transcription factor osterix(SP7)-binding sites on the LRP5 promoter and its pivotal role in upregulating LRP5 expression during implant osseointegration.However,the potential role of SP7 in ameliorating LRP5-dependent osteoporosis remained unknown.In this study,we used mice with a conditional knockout(c KO)of LRP5 in mature osteoblasts,which presented decreased osteogenesis.The in vitro experimental results showed that SP7 could promote LRP5 expression,thereby upregulating the osteogenic markers such as alkaline phosphatase(ALP),Runt-related transcription factor 2(Runx2),andβ-catenin(P<0.05).For the in vivo experiment,the SP7 overexpression virus was injected into a bone defect model of LRP5 c KO mice,resulting in increased bone mineral density(BMD)(P<0.001)and volumetric density(bone volume(BV)/total volume(TV))(P<0.001),and decreased trabecular separation(Tb.Sp)(P<0.05).These data suggested that SP7 could ameliorate bone defect healing in LRP5 c KO mice.Our study provides new insights into potential therapeutic opportunities for ameliorating LRP5-dependent osteoporosis.展开更多
目的鉴定乳腺癌干细胞系MCF-7中是否包含肿瘤干细胞相关的SP(side popu lation)亚群。方法制备MCF-7细胞悬液,经Hoechst33342和PI染色(其中一组同时加入拮抗剂维拉帕米作为对照),流式细胞仪分析SP亚群。另取贴壁生长的MCF-7细胞,用Hoech...目的鉴定乳腺癌干细胞系MCF-7中是否包含肿瘤干细胞相关的SP(side popu lation)亚群。方法制备MCF-7细胞悬液,经Hoechst33342和PI染色(其中一组同时加入拮抗剂维拉帕米作为对照),流式细胞仪分析SP亚群。另取贴壁生长的MCF-7细胞,用Hoechst33342和羟基荧光素二醋酸盐琥珀酰亚胺脂(CFSE)原位染色,荧光显微镜下观察细胞染色情况。结果SP亚群占MCF-7细胞的4%,维拉帕米阻断后减少为0.5%。荧光显微镜下见Hoechst33342阳性细胞约占95%。结论人乳腺癌细胞系MCF-7中存在SP亚群,即提示乳腺癌干细胞的存在;维拉帕米可抑制染料外排而减少SP细胞的比例。展开更多
用硅胶柱色谱及重结晶等方法,并根据理化性质和光谱数据从一株红树林共生真菌Paecilomycessp.Tree 1-7中分离鉴定了四个化合物,分别为secalon ic ac id A(1),tenellic ac id A(2),大黄素(3),大黄酚(4)。并首次对化合物1和2进行了抗人的...用硅胶柱色谱及重结晶等方法,并根据理化性质和光谱数据从一株红树林共生真菌Paecilomycessp.Tree 1-7中分离鉴定了四个化合物,分别为secalon ic ac id A(1),tenellic ac id A(2),大黄素(3),大黄酚(4)。并首次对化合物1和2进行了抗人的肝癌细胞hepG2的活性测试。结果显示,化合物1有很强的抗肿瘤活性,IC50=2.0μg/m l,化合物2的抗肿瘤活性IC50=62.1μg/m l。展开更多
目的体外培养HepG2细胞观察川西獐牙菜醇提物(Swertia mussitii Franch)对多药耐药相关蛋白3(multidrug resistance protein 3,MRP3)、核转录因子SP1(specificity protein 1 transcription factor,SP1)及核受体孕烷X受体(pregnane X rec...目的体外培养HepG2细胞观察川西獐牙菜醇提物(Swertia mussitii Franch)对多药耐药相关蛋白3(multidrug resistance protein 3,MRP3)、核转录因子SP1(specificity protein 1 transcription factor,SP1)及核受体孕烷X受体(pregnane X receptor,PXR)、CYP7A启动子结合因子(CYP7A promoter-binding factor,CPF)表达的影响。方法用MTT比色法检测川西獐牙菜醇提物的最佳作用浓度,再分别于0、12、24、48、72h刺激HepG2细胞,抽提细胞的RNA、总蛋白和核蛋白,采用荧光定量PCR和蛋白免疫印迹技术检测膜转运蛋白MRP3、转录因子SP1及核受体PXR、CPF在转录与蛋白水平的表达变化。每个时间点设立阴性对照DMSO组和阳性对照熊去氧胆酸(ursodeoxycholic acid,UDCA)组。结果川西獐牙菜醇提物可显著诱导HepG2细胞膜转运蛋白MRP3的mRNA和蛋白水平的高表达,其作用强于UDCA(P<0.05)。川西獐牙菜醇提物也可明显上调核转录因子SP1及核受体PXR的mRNA和蛋白表达水平,作用强于UDCA。对CPF表达的上调作用与UDCA相当(P<0.05)。结论川西獐牙菜醇提物可刺激HepG2细胞膜转运蛋白MRP3上调,且有可能是通过核转录因子SP1及核受体PXR、CPF上调其表达。展开更多
文摘Loss-of-function variants of low-density lipoprotein receptor-related protein 5(LRP5)can lead to reduced bone formation,culminating in diminished bone mass.Our previous study reported transcription factor osterix(SP7)-binding sites on the LRP5 promoter and its pivotal role in upregulating LRP5 expression during implant osseointegration.However,the potential role of SP7 in ameliorating LRP5-dependent osteoporosis remained unknown.In this study,we used mice with a conditional knockout(c KO)of LRP5 in mature osteoblasts,which presented decreased osteogenesis.The in vitro experimental results showed that SP7 could promote LRP5 expression,thereby upregulating the osteogenic markers such as alkaline phosphatase(ALP),Runt-related transcription factor 2(Runx2),andβ-catenin(P<0.05).For the in vivo experiment,the SP7 overexpression virus was injected into a bone defect model of LRP5 c KO mice,resulting in increased bone mineral density(BMD)(P<0.001)and volumetric density(bone volume(BV)/total volume(TV))(P<0.001),and decreased trabecular separation(Tb.Sp)(P<0.05).These data suggested that SP7 could ameliorate bone defect healing in LRP5 c KO mice.Our study provides new insights into potential therapeutic opportunities for ameliorating LRP5-dependent osteoporosis.
文摘目的鉴定乳腺癌干细胞系MCF-7中是否包含肿瘤干细胞相关的SP(side popu lation)亚群。方法制备MCF-7细胞悬液,经Hoechst33342和PI染色(其中一组同时加入拮抗剂维拉帕米作为对照),流式细胞仪分析SP亚群。另取贴壁生长的MCF-7细胞,用Hoechst33342和羟基荧光素二醋酸盐琥珀酰亚胺脂(CFSE)原位染色,荧光显微镜下观察细胞染色情况。结果SP亚群占MCF-7细胞的4%,维拉帕米阻断后减少为0.5%。荧光显微镜下见Hoechst33342阳性细胞约占95%。结论人乳腺癌细胞系MCF-7中存在SP亚群,即提示乳腺癌干细胞的存在;维拉帕米可抑制染料外排而减少SP细胞的比例。
文摘用硅胶柱色谱及重结晶等方法,并根据理化性质和光谱数据从一株红树林共生真菌Paecilomycessp.Tree 1-7中分离鉴定了四个化合物,分别为secalon ic ac id A(1),tenellic ac id A(2),大黄素(3),大黄酚(4)。并首次对化合物1和2进行了抗人的肝癌细胞hepG2的活性测试。结果显示,化合物1有很强的抗肿瘤活性,IC50=2.0μg/m l,化合物2的抗肿瘤活性IC50=62.1μg/m l。
文摘目的体外培养HepG2细胞观察川西獐牙菜醇提物(Swertia mussitii Franch)对多药耐药相关蛋白3(multidrug resistance protein 3,MRP3)、核转录因子SP1(specificity protein 1 transcription factor,SP1)及核受体孕烷X受体(pregnane X receptor,PXR)、CYP7A启动子结合因子(CYP7A promoter-binding factor,CPF)表达的影响。方法用MTT比色法检测川西獐牙菜醇提物的最佳作用浓度,再分别于0、12、24、48、72h刺激HepG2细胞,抽提细胞的RNA、总蛋白和核蛋白,采用荧光定量PCR和蛋白免疫印迹技术检测膜转运蛋白MRP3、转录因子SP1及核受体PXR、CPF在转录与蛋白水平的表达变化。每个时间点设立阴性对照DMSO组和阳性对照熊去氧胆酸(ursodeoxycholic acid,UDCA)组。结果川西獐牙菜醇提物可显著诱导HepG2细胞膜转运蛋白MRP3的mRNA和蛋白水平的高表达,其作用强于UDCA(P<0.05)。川西獐牙菜醇提物也可明显上调核转录因子SP1及核受体PXR的mRNA和蛋白表达水平,作用强于UDCA。对CPF表达的上调作用与UDCA相当(P<0.05)。结论川西獐牙菜醇提物可刺激HepG2细胞膜转运蛋白MRP3上调,且有可能是通过核转录因子SP1及核受体PXR、CPF上调其表达。
