活化T细胞核因子(nuclear factor-activated T cell 1,NFATc1)是一种重要的转录因子。在破骨细胞中,它由上游RANKL信号通路的诱导、Ca^(2+)相关协同刺激信号通路与Ca^(2+)非依赖信号通路的扩增,Lhx2、IRF8、Mafb及Bcl6等细胞因子负反馈...活化T细胞核因子(nuclear factor-activated T cell 1,NFATc1)是一种重要的转录因子。在破骨细胞中,它由上游RANKL信号通路的诱导、Ca^(2+)相关协同刺激信号通路与Ca^(2+)非依赖信号通路的扩增,Lhx2、IRF8、Mafb及Bcl6等细胞因子负反馈诱导,在NFATc1转录过程中的启动、扩增及靶向作用三个阶段通过复杂交互的调节影响其下游各种靶基因及蛋白,最终介导破骨细胞的分化、融合及对无机和有机骨基质的降解作用。宏观上,NFATc1还受到外界机械应力的影响从而在破骨细胞生长过程中发挥作用;并且NFATc1的调节过程受其自身节律的影响。本文就NFATc1的结构、相关调节机制和对破骨细胞的作用研究进展进行综述。展开更多
背景:研究发现miR-25的下调与活化T细胞核因子c1(nuclear factor of activated T cells c1,NFATc1)过度激活相关,而NFATc1是破骨细胞分化成熟中的核心转录因子,提示miR-25的表达水平能调控破骨细胞分化。目的:探讨过表达miR-25对Ti颗粒...背景:研究发现miR-25的下调与活化T细胞核因子c1(nuclear factor of activated T cells c1,NFATc1)过度激活相关,而NFATc1是破骨细胞分化成熟中的核心转录因子,提示miR-25的表达水平能调控破骨细胞分化。目的:探讨过表达miR-25对Ti颗粒诱导的破骨细胞分化的影响,并探索其中的信号通路。方法:体外培养RAW264.7细胞系,CCK-8测定Ti颗粒对细胞增殖的影响;转染miR-25模拟物(mimics)至RAW264.7细胞,并与Ti颗粒培养诱导破骨细胞的分化;免疫荧光检测NFATc1的激活状态,TRAP染色及计数验证各组破骨细胞分化情况,实时荧光定量PCR检测Ca2+/NFATc1通路分子NFATc1、CaMKⅡ、CaMKⅣmRNA表达。结果与结论:(1)低浓度的Ti颗粒(0.1 g/L)对细胞增殖无明显影响;(2)与阴性对照组相比,miR-25过表达组破骨细胞(TRAP阳性细胞)生成明显减少(P <0.05);(3)细胞内转染miR-25模拟物后,免疫荧光检测见NFATc1的激活减少,荧光强度降低;(4)miR-25过表达后信号通路分子NFATc1、CaMKⅡ、CaMKⅣmRNA的表达显著下降(P <0.05)。(5)结果表明,miR-25过表达能抑制Ti颗粒诱导的破骨细胞分化,这一过程可能是通过Ca2+/NFATc1通路实现的。展开更多
PIP5k1βis crucial to the generation of phosphotidylinosotol(4,5)P2.PIP5k1βparticipates in numerous cellular activities,such as B cell and platelet activation,cell phagocytosis and endocytosis,cell apoptosis,and cyto...PIP5k1βis crucial to the generation of phosphotidylinosotol(4,5)P2.PIP5k1βparticipates in numerous cellular activities,such as B cell and platelet activation,cell phagocytosis and endocytosis,cell apoptosis,and cytoskeletal organization.In the present work,we aimed to examine the function of PIP5k1βin osteoclastogenesis and osteogenesis to provide promising strategies for osteoporosis prevention and treatment.We discovered that PIP5k1β deletion in mice resulted in obvious bone loss and that PIP5k1β was highly expressed during both osteoclast and osteoblast differentiation.Deletion of the gene was found to enhance the proliferation and migration of bone marrow-derived macrophage-like cells to promote osteoclast differentiation.PIP5k1β-/-osteoclasts exhibited normal cytoskeleton architecture but stronger resorption activity.PIP5kip deficiency also promoted activation of mitogen-activated kinase and Akt signaling,enhanced TRAF6 and c-Fos expression,facilitated the expression and nuclear translocation of NFATC1,and upregulated Grb2 expression,thereby accelerating osteoclast differentiation and function.Finally,PIP5k1β enhanced osteoblast differentiation by upregulating master gene expression through triggering smad1/5/8 signaling.Therefore,PIP5k1βmodulates bone homeostasis and remodeling.展开更多
目的:探讨胞浆活化T细胞核因子1(nuclear factor of activated T-cells,cytplasmic 1,NFATc1)对人卵巢癌SKOV3细胞裸鼠移植瘤生长和肿瘤脉管生成的影响及其可能机制。方法:NFATc1 si RNA转染人上皮性卵巢癌细胞株SKOV3,免疫荧光及RT-PC...目的:探讨胞浆活化T细胞核因子1(nuclear factor of activated T-cells,cytplasmic 1,NFATc1)对人卵巢癌SKOV3细胞裸鼠移植瘤生长和肿瘤脉管生成的影响及其可能机制。方法:NFATc1 si RNA转染人上皮性卵巢癌细胞株SKOV3,免疫荧光及RT-PCR测量转染效率和基因抑制率,选取效率最高的序列建立裸鼠皮下移植瘤模型,测量各组裸鼠肿瘤体积,观察NFATc1 siRNA的体内抗肿瘤作用。免疫组织化学检测各组肿瘤组织NFATc1的表达情况,并使用细胞角蛋白染色标记上皮性来源,CD34标记微血管,podoplanin标记微淋巴管。分别计算各组微血管及微淋巴管密度并进行统计学分析。应用RT-PCR及Western blot检测各组移植瘤组织NFATc1、CXC趋化因子受体2(CXCR2)、成纤维细胞生长因子2(FGF-2)及血小板源性生长因子BB(PDGF-BB)的mRNA及蛋白表达水平。结果:3条特异性序列均可显著降低NFATc1的表达水平,以siRNA-1169最佳。NFATc1在空白组及阴性对照组瘤组织高表达。干扰组抑瘤率为57.08%,且重量和体积均低于2个对照组。空白组和阴性对照组的微血管密度和微淋巴管密度明显高于干扰组。对照组比较,NFATc1 siRNA可以在mRNA水平上明显抑制NFATC1、CXCR2、FGF-2和PDGF-BB的转录。Western blot各组细胞在相应位置出现NFATc1、CXCR2、FGF-2和PDGF-BB条带,空白组与阴性对照组的吸光度最强,与干扰组比较具有显著差异。结论:NFATc1 siRNA明显抑制人卵巢癌SKOV3细胞裸鼠皮下移植瘤生长和肿瘤脉管生成,下调CXCR2、FGF-2及PDGF-BB的表达可能为其途径之一。展开更多
Inflammation can initiate osteolysis,which is the breakdown of bone by fully developed osteoclasts.The com-pound Oleandrin is recognized for its effects against inflammation and tumors.Our objective was to examine the...Inflammation can initiate osteolysis,which is the breakdown of bone by fully developed osteoclasts.The com-pound Oleandrin is recognized for its effects against inflammation and tumors.Our objective was to examine the effects of Oleandrin on osteoclastogenesis and osteolysis,both in vitro and in vivo.In vitro,the impact of Oleandrin on osteoclastogenesis was assessed using CCK-8 assays,TRAP staining,and bone resorption assays.Ad-ditionally,a mouse model of osteolysis caused by LPS injection into the calvaria was used to conduct an in vivo investigation,examining bone histomorphology,histology,and immunohistochemistry.In vitro,concentrations of 5 nM and 10 nM of Oleandrin were found to be non-cytotoxic based on the results obtained.In vitro,Olean-drin hindered the osteoclastogenesis and bone resorption induced by RANKL.Oleandrin successfully inhibited the phosphorylation of NF-κB p65 and PI3K p85 in osteolytic tissue,thereby suppressing LPS-induced inflammatory osteolysis in mice calvaria during the in vivo study.Furthermore,the Oleandrin-treated group exhibited a note-worthy decrease in the expression level of NFATc1,which is a crucial controller of osteoclastogenesis.To sum up,our discoveries indicate that Oleandrin could hinder osteoclastogenesis and bone resorption,thereby having the ability to suppress inflammation-induced osteolysis.The underlying mechanism involves the NF-κB/PI3K pathway and inhibition of NFATc1 activation.Therefore,the findings suggest that Oleandrin holds potential as a therapeutic remedy for osteolytic ailments.展开更多
目的研究桂皮醛对激素诱导的体外破骨细胞增殖分化及骨吸收功能增强的保护作用及分子机制。方法RANKL联合M-CSF体外诱导RAW264.7细胞分化为破骨细胞,分为对照组、地塞米松处理组和不同浓度(11.6、23.2、46.4μg·L-1)桂皮醛(cinnami...目的研究桂皮醛对激素诱导的体外破骨细胞增殖分化及骨吸收功能增强的保护作用及分子机制。方法RANKL联合M-CSF体外诱导RAW264.7细胞分化为破骨细胞,分为对照组、地塞米松处理组和不同浓度(11.6、23.2、46.4μg·L-1)桂皮醛(cinnamic alde byde,CA)干预组;TRAP染色试剂盒鉴定成熟破骨细胞;MTT法观察地塞米松和桂皮醛在不同时间点对破骨细胞增殖的抑制作用;ELISA法检测细胞培养液上清中TRACP5b的表达水平;RT-PCR分析破骨细胞相关转录因子(RANK、NFATc1)mRNA的表达状态。结果地塞米松处理后,破骨细胞的增殖分化及骨吸收功能明显增强(P<0.05);而经不同浓度桂皮醛干预后,与地塞米松组相比,细胞增殖活性、上清液中TRACP-5b的含量以及RANK、NFATc1mRNA的表达水平随药物浓度的增加呈剂量依赖性降低(P<0.05)。结论桂皮醛可有效抑制激素诱导的破骨细胞的增殖及骨吸收功能,其作用机制可能是通过下调RANK以及下游NFATc1基因的表达。展开更多
文摘活化T细胞核因子(nuclear factor-activated T cell 1,NFATc1)是一种重要的转录因子。在破骨细胞中,它由上游RANKL信号通路的诱导、Ca^(2+)相关协同刺激信号通路与Ca^(2+)非依赖信号通路的扩增,Lhx2、IRF8、Mafb及Bcl6等细胞因子负反馈诱导,在NFATc1转录过程中的启动、扩增及靶向作用三个阶段通过复杂交互的调节影响其下游各种靶基因及蛋白,最终介导破骨细胞的分化、融合及对无机和有机骨基质的降解作用。宏观上,NFATc1还受到外界机械应力的影响从而在破骨细胞生长过程中发挥作用;并且NFATc1的调节过程受其自身节律的影响。本文就NFATc1的结构、相关调节机制和对破骨细胞的作用研究进展进行综述。
文摘背景:研究发现miR-25的下调与活化T细胞核因子c1(nuclear factor of activated T cells c1,NFATc1)过度激活相关,而NFATc1是破骨细胞分化成熟中的核心转录因子,提示miR-25的表达水平能调控破骨细胞分化。目的:探讨过表达miR-25对Ti颗粒诱导的破骨细胞分化的影响,并探索其中的信号通路。方法:体外培养RAW264.7细胞系,CCK-8测定Ti颗粒对细胞增殖的影响;转染miR-25模拟物(mimics)至RAW264.7细胞,并与Ti颗粒培养诱导破骨细胞的分化;免疫荧光检测NFATc1的激活状态,TRAP染色及计数验证各组破骨细胞分化情况,实时荧光定量PCR检测Ca2+/NFATc1通路分子NFATc1、CaMKⅡ、CaMKⅣmRNA表达。结果与结论:(1)低浓度的Ti颗粒(0.1 g/L)对细胞增殖无明显影响;(2)与阴性对照组相比,miR-25过表达组破骨细胞(TRAP阳性细胞)生成明显减少(P <0.05);(3)细胞内转染miR-25模拟物后,免疫荧光检测见NFATc1的激活减少,荧光强度降低;(4)miR-25过表达后信号通路分子NFATc1、CaMKⅡ、CaMKⅣmRNA的表达显著下降(P <0.05)。(5)结果表明,miR-25过表达能抑制Ti颗粒诱导的破骨细胞分化,这一过程可能是通过Ca2+/NFATc1通路实现的。
基金This work was supported by grants from the National Natural Science Foundation of China(81830078,81772347,and 81572123)Science and Technology Commission of Shanghai Municipality(19XD1434100 and 16430723500)+1 种基金Shanghai Municipal Education Commission-Gaofeng Clinical Medicine Grant Support(2016131A)Shanghai Jiao Tong University-The Chinese University of Hong Kong joint Research Collaboration Fund.
文摘PIP5k1βis crucial to the generation of phosphotidylinosotol(4,5)P2.PIP5k1βparticipates in numerous cellular activities,such as B cell and platelet activation,cell phagocytosis and endocytosis,cell apoptosis,and cytoskeletal organization.In the present work,we aimed to examine the function of PIP5k1βin osteoclastogenesis and osteogenesis to provide promising strategies for osteoporosis prevention and treatment.We discovered that PIP5k1β deletion in mice resulted in obvious bone loss and that PIP5k1β was highly expressed during both osteoclast and osteoblast differentiation.Deletion of the gene was found to enhance the proliferation and migration of bone marrow-derived macrophage-like cells to promote osteoclast differentiation.PIP5k1β-/-osteoclasts exhibited normal cytoskeleton architecture but stronger resorption activity.PIP5kip deficiency also promoted activation of mitogen-activated kinase and Akt signaling,enhanced TRAF6 and c-Fos expression,facilitated the expression and nuclear translocation of NFATC1,and upregulated Grb2 expression,thereby accelerating osteoclast differentiation and function.Finally,PIP5k1β enhanced osteoblast differentiation by upregulating master gene expression through triggering smad1/5/8 signaling.Therefore,PIP5k1βmodulates bone homeostasis and remodeling.
基金National Natural Science Foundation of China(82272165)Shanghai Municipal Commission of Health and Family Planning’s Science and Research Fund(202040141)+2 种基金Shanghai Municipality Science and Technology Commission(20ZR1451800,22ZR1457200)Shanghai Municipal Health Commission(2022YQ006)Shanghai Tongren Hospital(TRKYRC-xx202203).
文摘Inflammation can initiate osteolysis,which is the breakdown of bone by fully developed osteoclasts.The com-pound Oleandrin is recognized for its effects against inflammation and tumors.Our objective was to examine the effects of Oleandrin on osteoclastogenesis and osteolysis,both in vitro and in vivo.In vitro,the impact of Oleandrin on osteoclastogenesis was assessed using CCK-8 assays,TRAP staining,and bone resorption assays.Ad-ditionally,a mouse model of osteolysis caused by LPS injection into the calvaria was used to conduct an in vivo investigation,examining bone histomorphology,histology,and immunohistochemistry.In vitro,concentrations of 5 nM and 10 nM of Oleandrin were found to be non-cytotoxic based on the results obtained.In vitro,Olean-drin hindered the osteoclastogenesis and bone resorption induced by RANKL.Oleandrin successfully inhibited the phosphorylation of NF-κB p65 and PI3K p85 in osteolytic tissue,thereby suppressing LPS-induced inflammatory osteolysis in mice calvaria during the in vivo study.Furthermore,the Oleandrin-treated group exhibited a note-worthy decrease in the expression level of NFATc1,which is a crucial controller of osteoclastogenesis.To sum up,our discoveries indicate that Oleandrin could hinder osteoclastogenesis and bone resorption,thereby having the ability to suppress inflammation-induced osteolysis.The underlying mechanism involves the NF-κB/PI3K pathway and inhibition of NFATc1 activation.Therefore,the findings suggest that Oleandrin holds potential as a therapeutic remedy for osteolytic ailments.
文摘目的研究桂皮醛对激素诱导的体外破骨细胞增殖分化及骨吸收功能增强的保护作用及分子机制。方法RANKL联合M-CSF体外诱导RAW264.7细胞分化为破骨细胞,分为对照组、地塞米松处理组和不同浓度(11.6、23.2、46.4μg·L-1)桂皮醛(cinnamic alde byde,CA)干预组;TRAP染色试剂盒鉴定成熟破骨细胞;MTT法观察地塞米松和桂皮醛在不同时间点对破骨细胞增殖的抑制作用;ELISA法检测细胞培养液上清中TRACP5b的表达水平;RT-PCR分析破骨细胞相关转录因子(RANK、NFATc1)mRNA的表达状态。结果地塞米松处理后,破骨细胞的增殖分化及骨吸收功能明显增强(P<0.05);而经不同浓度桂皮醛干预后,与地塞米松组相比,细胞增殖活性、上清液中TRACP-5b的含量以及RANK、NFATc1mRNA的表达水平随药物浓度的增加呈剂量依赖性降低(P<0.05)。结论桂皮醛可有效抑制激素诱导的破骨细胞的增殖及骨吸收功能,其作用机制可能是通过下调RANK以及下游NFATc1基因的表达。
