目的:探讨红景天苷(SAL)对低氧条件下脂多糖(LPS)诱导的人牙周膜细胞(h PDLCs)表达活性氧(ROS)及炎症因子的影响,并初步探讨其相关作用机制。方法:首先用含SAL终浓度为0、30、60、100μg/m L的DMEM分别培养第5代h PDLCs,并于培养后1、2...目的:探讨红景天苷(SAL)对低氧条件下脂多糖(LPS)诱导的人牙周膜细胞(h PDLCs)表达活性氧(ROS)及炎症因子的影响,并初步探讨其相关作用机制。方法:首先用含SAL终浓度为0、30、60、100μg/m L的DMEM分别培养第5代h PDLCs,并于培养后1、2、3、4、5、6 d各时间点,采用MTT法检测各浓度组细胞的增殖活性。然后再取第5代h PDLCs并将其随机分为5组,分别用不含LPS和SAL的DMEM(对照组)以及含10μg/m L LPS(单纯LPS组)、10μg/m L LPS+30μg/m L SAL、10μg/m L LPS+60μg/m L SAL、10μg/m L LPS+100μg/m L SAL的DMEM进行低氧(10 m L/L O2)培养;连续培养6 h后,收集各组细胞及其上清液,分别用DCFH-DA法检测各组细胞中ROS的表达水平、RT-PCR和ELISA法检测各组细胞中TNF-α、IL-1β、IL-6的m RNA和蛋白的表达水平、Western-blot法检测对照组、单纯LPS组和10μg/m L LPS+100μg/m L SAL细胞的核转录因子-κB(NF-κB)信号通路的磷酸化水平。结果:30、60、100μg/m L SAL对h PDLCs的增殖无明显影响(P>0.05);上述3个浓度SAL均能明显降低LPS诱导的h PDLCs中ROS水平以及TNF-α、IL-1β、IL-6 m RNA和蛋白的表达水平(P<0.05);100μg/m L SAL可明显降低LPS诱导的h PDLCs中NF-κB信号通路的磷酸化水平(P<0.05)。结论:SAL可能通过抑制NF-κB信号通路的激活,降低低氧条件下LPS诱导的h PDLCs中活性氧及炎症因子的表达水平。展开更多
目的:观察静压力对人牙周膜干细胞(h PDLSCs)骨向分化能力的影响。方法:体外培养h PDLSCs,并将其随机分为4个组;置于压力加载装置内分别给予0(对照组)、20、100、200 k Pa的静压力刺激,连续加压6 h后分别采用Quantitative RT-PCR和Weste...目的:观察静压力对人牙周膜干细胞(h PDLSCs)骨向分化能力的影响。方法:体外培养h PDLSCs,并将其随机分为4个组;置于压力加载装置内分别给予0(对照组)、20、100、200 k Pa的静压力刺激,连续加压6 h后分别采用Quantitative RT-PCR和Western-blots法检测h PDLSCs的破骨细胞核因子KB受体活化因子配基(RANKL)及骨保护因子(OPG)的表达。结果:与对照组相比,当压力值为20 k Pa时,RANKL mRNA和蛋白的表达量变化不明显(P>0.05),但OPG的表达量明显升高(P<0.05),RANKL/OPG的比值明显降低(P<0.05);当压力值为100 k Pa时,RANKL mRNA和蛋白的表达量明显升高(P<0.05),但OPG的表达量降低(P<0.05),RANKL/OPG的比值明显升高(P<0.05);当压力值为200 k Pa时,RANKL、OPG mRNA和蛋白的表达量均明显降低(P<0.05),其中以RANKL下降的程度更加明显,RANKL/OPG的比值明显降低(P<0.05)。结论:持续的静压力作用可使h PDLSCs表达OPG和RANKL的水平发生明显变化,并具有力值依赖性。展开更多
Periodontitis is a type of chronic inflammation in the gingival tissue caused by infectious bacteria colonizing the surface of the teeth,leading to the destruction of tooth-supporting tissues and loss of alveolar bone...Periodontitis is a type of chronic inflammation in the gingival tissue caused by infectious bacteria colonizing the surface of the teeth,leading to the destruction of tooth-supporting tissues and loss of alveolar bone.Suberoylanilide hydroxamic acid(SAHA),a class of histone deacetylase(HDAC)inhibitor,has the potential to stimulate osteoblast differentiation by acetylating histone proteins,and thus suppressing the expression of adipogenic and chondrogenic genes.However,the effect of SAHA on the differentiation of human periodontal ligament stem cells(hPDLSCs)is yet to be elucidated.Herein,we investigated the effects of SAHA on in vitro proliferation and differentiation of hPDLSCs by MTT assay,Alizarin Red-S,and alkaline phosphatase staining,and real-time PCR.Notably,300 ng/mL SAHA treatment enhanced the proliferation and mineralization of hPDLSCs,indicating their osteogenic potential.Moreover,a significant enhancement of osteogenesis gene markers and proteins was observed.We also demonstrated that ERK is a positive regulator of Runx2 transcription factors during osteoblast differentiation.These results indicate that SAHA may be a useful osteogenic induction agent for periodontal bone regeneration.展开更多
文摘目的:探讨红景天苷(SAL)对低氧条件下脂多糖(LPS)诱导的人牙周膜细胞(h PDLCs)表达活性氧(ROS)及炎症因子的影响,并初步探讨其相关作用机制。方法:首先用含SAL终浓度为0、30、60、100μg/m L的DMEM分别培养第5代h PDLCs,并于培养后1、2、3、4、5、6 d各时间点,采用MTT法检测各浓度组细胞的增殖活性。然后再取第5代h PDLCs并将其随机分为5组,分别用不含LPS和SAL的DMEM(对照组)以及含10μg/m L LPS(单纯LPS组)、10μg/m L LPS+30μg/m L SAL、10μg/m L LPS+60μg/m L SAL、10μg/m L LPS+100μg/m L SAL的DMEM进行低氧(10 m L/L O2)培养;连续培养6 h后,收集各组细胞及其上清液,分别用DCFH-DA法检测各组细胞中ROS的表达水平、RT-PCR和ELISA法检测各组细胞中TNF-α、IL-1β、IL-6的m RNA和蛋白的表达水平、Western-blot法检测对照组、单纯LPS组和10μg/m L LPS+100μg/m L SAL细胞的核转录因子-κB(NF-κB)信号通路的磷酸化水平。结果:30、60、100μg/m L SAL对h PDLCs的增殖无明显影响(P>0.05);上述3个浓度SAL均能明显降低LPS诱导的h PDLCs中ROS水平以及TNF-α、IL-1β、IL-6 m RNA和蛋白的表达水平(P<0.05);100μg/m L SAL可明显降低LPS诱导的h PDLCs中NF-κB信号通路的磷酸化水平(P<0.05)。结论:SAL可能通过抑制NF-κB信号通路的激活,降低低氧条件下LPS诱导的h PDLCs中活性氧及炎症因子的表达水平。
文摘目的:观察静压力对人牙周膜干细胞(h PDLSCs)骨向分化能力的影响。方法:体外培养h PDLSCs,并将其随机分为4个组;置于压力加载装置内分别给予0(对照组)、20、100、200 k Pa的静压力刺激,连续加压6 h后分别采用Quantitative RT-PCR和Western-blots法检测h PDLSCs的破骨细胞核因子KB受体活化因子配基(RANKL)及骨保护因子(OPG)的表达。结果:与对照组相比,当压力值为20 k Pa时,RANKL mRNA和蛋白的表达量变化不明显(P>0.05),但OPG的表达量明显升高(P<0.05),RANKL/OPG的比值明显降低(P<0.05);当压力值为100 k Pa时,RANKL mRNA和蛋白的表达量明显升高(P<0.05),但OPG的表达量降低(P<0.05),RANKL/OPG的比值明显升高(P<0.05);当压力值为200 k Pa时,RANKL、OPG mRNA和蛋白的表达量均明显降低(P<0.05),其中以RANKL下降的程度更加明显,RANKL/OPG的比值明显降低(P<0.05)。结论:持续的静压力作用可使h PDLSCs表达OPG和RANKL的水平发生明显变化,并具有力值依赖性。
基金the National Research Foundation of Korea(NRF)and funded by the Ministry of Education(No.2018R1A6A1A03025582)the National Research Foundation of Korea(NRF-2019R1D1A3A03103828).
文摘Periodontitis is a type of chronic inflammation in the gingival tissue caused by infectious bacteria colonizing the surface of the teeth,leading to the destruction of tooth-supporting tissues and loss of alveolar bone.Suberoylanilide hydroxamic acid(SAHA),a class of histone deacetylase(HDAC)inhibitor,has the potential to stimulate osteoblast differentiation by acetylating histone proteins,and thus suppressing the expression of adipogenic and chondrogenic genes.However,the effect of SAHA on the differentiation of human periodontal ligament stem cells(hPDLSCs)is yet to be elucidated.Herein,we investigated the effects of SAHA on in vitro proliferation and differentiation of hPDLSCs by MTT assay,Alizarin Red-S,and alkaline phosphatase staining,and real-time PCR.Notably,300 ng/mL SAHA treatment enhanced the proliferation and mineralization of hPDLSCs,indicating their osteogenic potential.Moreover,a significant enhancement of osteogenesis gene markers and proteins was observed.We also demonstrated that ERK is a positive regulator of Runx2 transcription factors during osteoblast differentiation.These results indicate that SAHA may be a useful osteogenic induction agent for periodontal bone regeneration.
