Osteoporosis is a prevalent skeletal condition characterized by reduced bone mass and strength,leading to increased fragility.Buqi-Tongluo(BQTL)decoction,a traditional Chinese medicine(TCM)prescription,has yet to be f...Osteoporosis is a prevalent skeletal condition characterized by reduced bone mass and strength,leading to increased fragility.Buqi-Tongluo(BQTL)decoction,a traditional Chinese medicine(TCM)prescription,has yet to be fully evaluated for its potential in treating bone diseases such as osteoporosis.To investigate the mechanism by which BQTL decoction inhibits osteoclast differentiation in vitro and validate these findings through in vivo experiments.We employed MTS assays to assess the potential proliferative or toxic effects of BQTL on bone marrow macrophages(BMMs)at various concentrations.TRAc P experiments were conducted to examine BQTL's impact on osteoclast differentiation.RT-PCR and Western blot analyses were utilized to evaluate the relative expression levels of osteoclast-specific genes and proteins under BQTL stimulation.Finally,in vivo experiments were performed using an osteoporosis model to further validate the in vitro findings.This study revealed that BQTL suppressed receptor activator of NF-κB ligand(RANKL)-induced osteoclastogenesis and osteoclast resorption activity in vitro in a dose-dependent manner without observable cytotoxicity.The inhibitory effects of BQTL on osteoclast formation and function were attributed to the downregulation of NFATc1 and c-fos activity,primarily through attenuation of the MAPK,NF-κB,and Calcineurin signaling pathways.BQTL's inhibitory capacity was further examined in vivo using an ovariectomized(OVX)rat model,demonstrating a strong protective effect against bone loss.BQTL may serve as an effective therapeutic TCM for the treatment of postmenopausal osteoporosis and the alleviation of bone loss induced by estrogen deficiency and related conditions.展开更多
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
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
基金supported by the Natural Science Foundation of Guangdong Province,China(No.2021A1515012168)the Administration of Traditional Chinese Medicine of Guangdong Province,China(Nos.20221146 and 20241091)+5 种基金the Basic and Applied Basic Research Fund Project in Guangdong Province,China(No.2020A1515110948)the Basic and Applied Basic Research in Jointly Funded Projects of City Schools(Institutes)Projects,China(Nos.202201020500 and 202201020295)the Project of Guangzhou Science and Technology Department,China(No.202102021040)the Guangzhou Science and Technology Plan Project,China(No.2023B03J0379)the Chinese Society of Traditional Chinese Medicine Youth Talent Lifting Project(No.2022-QNRC2-B11)the Hospital Young and Middle-aged Key Talent Cultivation Project of The First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine(2023-10)。
文摘Osteoporosis is a prevalent skeletal condition characterized by reduced bone mass and strength,leading to increased fragility.Buqi-Tongluo(BQTL)decoction,a traditional Chinese medicine(TCM)prescription,has yet to be fully evaluated for its potential in treating bone diseases such as osteoporosis.To investigate the mechanism by which BQTL decoction inhibits osteoclast differentiation in vitro and validate these findings through in vivo experiments.We employed MTS assays to assess the potential proliferative or toxic effects of BQTL on bone marrow macrophages(BMMs)at various concentrations.TRAc P experiments were conducted to examine BQTL's impact on osteoclast differentiation.RT-PCR and Western blot analyses were utilized to evaluate the relative expression levels of osteoclast-specific genes and proteins under BQTL stimulation.Finally,in vivo experiments were performed using an osteoporosis model to further validate the in vitro findings.This study revealed that BQTL suppressed receptor activator of NF-κB ligand(RANKL)-induced osteoclastogenesis and osteoclast resorption activity in vitro in a dose-dependent manner without observable cytotoxicity.The inhibitory effects of BQTL on osteoclast formation and function were attributed to the downregulation of NFATc1 and c-fos activity,primarily through attenuation of the MAPK,NF-κB,and Calcineurin signaling pathways.BQTL's inhibitory capacity was further examined in vivo using an ovariectomized(OVX)rat model,demonstrating a strong protective effect against bone loss.BQTL may serve as an effective therapeutic TCM for the treatment of postmenopausal osteoporosis and the alleviation of bone loss induced by estrogen deficiency and related conditions.
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。
