Background and Objective: It has been found that human periodontal ligament (hPDL) cells express cannabinoid receptor CB2. However, the functional importance of CB2 in hPDL cells exposed to bacterial endotoxins is not...Background and Objective: It has been found that human periodontal ligament (hPDL) cells express cannabinoid receptor CB2. However, the functional importance of CB2 in hPDL cells exposed to bacterial endotoxins is not known. Here we investigate if the inflammation promoter lipopolysaccharide (LPS) affects CB2 expression and if activation of CB2 regulates LPS-induced pro-inflammatory cytokine production and osteoclastogenic gene expression in hPDL cells. Methods: The hPDL cells were obtained from extracted teeth of periodontally healthy subjects. CB2 expression in hPDL cells exposed to LPS was deter- mined by quantitative real-time PCR analysis. Then, the cells were incubated with or without CB2-specific agonist HU-308 before further stimulation with LPS. In some experiments, the cells were pre-treated with CB2-specific antagonist SR144528. The production of pro-inflammatory cytokines interleukin-1 beta (IL- 1β), interleukin-6 (IL-6) and tumor necrosis factoralpha (TNF-α) was assessed by enzyme-linked immunosorbent assay (ELISA). The mRNA expression of osteoclastogenic genes osteoprotegerin (OPG) and receptor activator of NF-κB ligand (RANKL) was examined using quantitative real-time PCR analysis. Results: CB2 expression in hPDL cells was markedly enhanced by LPS. HU-308 significantly suppressed the production of IL-1β, IL-6 and TNF-α exposed to LPS, whereas SR144528 attenuated this effect. The OPG/RANKL ratio decreased when exposed to LPS, furthermore increased significantly with the addition of HU-308 and finally decreased markedly after pretreatment with SR144528. Conclusion: Our study demonstrated that activation of CB2 had anti-inflammatory and anti-resorptive effects on LPS-stimulated hPDL cells. These findings suggest that activation of CB2 might be an effective therapeutic strategy for the treatment of inflammation and alveolar bone resorption in periodontitis.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
文摘Background and Objective: It has been found that human periodontal ligament (hPDL) cells express cannabinoid receptor CB2. However, the functional importance of CB2 in hPDL cells exposed to bacterial endotoxins is not known. Here we investigate if the inflammation promoter lipopolysaccharide (LPS) affects CB2 expression and if activation of CB2 regulates LPS-induced pro-inflammatory cytokine production and osteoclastogenic gene expression in hPDL cells. Methods: The hPDL cells were obtained from extracted teeth of periodontally healthy subjects. CB2 expression in hPDL cells exposed to LPS was deter- mined by quantitative real-time PCR analysis. Then, the cells were incubated with or without CB2-specific agonist HU-308 before further stimulation with LPS. In some experiments, the cells were pre-treated with CB2-specific antagonist SR144528. The production of pro-inflammatory cytokines interleukin-1 beta (IL- 1β), interleukin-6 (IL-6) and tumor necrosis factoralpha (TNF-α) was assessed by enzyme-linked immunosorbent assay (ELISA). The mRNA expression of osteoclastogenic genes osteoprotegerin (OPG) and receptor activator of NF-κB ligand (RANKL) was examined using quantitative real-time PCR analysis. Results: CB2 expression in hPDL cells was markedly enhanced by LPS. HU-308 significantly suppressed the production of IL-1β, IL-6 and TNF-α exposed to LPS, whereas SR144528 attenuated this effect. The OPG/RANKL ratio decreased when exposed to LPS, furthermore increased significantly with the addition of HU-308 and finally decreased markedly after pretreatment with SR144528. Conclusion: Our study demonstrated that activation of CB2 had anti-inflammatory and anti-resorptive effects on LPS-stimulated hPDL cells. These findings suggest that activation of CB2 might be an effective therapeutic strategy for the treatment of inflammation and alveolar bone resorption in periodontitis.
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。
