中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2014年
14期
1101-1104
,共4页
钟兴%冯颖瑜%苏磊%卫国红%潘天荣%修玲玲
鐘興%馮穎瑜%囌磊%衛國紅%潘天榮%脩玲玲
종흥%풍영유%소뢰%위국홍%반천영%수령령
成骨细胞%棕榈酸%NF-κB
成骨細胞%棕櫚痠%NF-κB
성골세포%종려산%NF-κB
Osteoblasts%Palmitic acid%NF-kappa B
目的:探讨棕榈酸诱导MC3T3-E1成骨细胞的凋亡是否与核因子( NF)-κB的激活有关。方法观察不同浓度(0~1000μmol/L)棕榈酸孵育MC3T3-E1成骨细胞24 h后对细胞活力和细胞凋亡的影响。细胞活力用四甲基偶氮唑盐法( MTT法)检测,细胞凋亡用Hochest 33258染核和Western印迹法检测活化型( cleaved ) caspase-3蛋白验证。用500μmol/L棕榈酸分别孵育细胞0~240 min,Western印迹法检测IκBα、p-NF-κB p65蛋白表达。结果棕榈酸呈时间和浓度依赖的方式抑制MC3T3-E1细胞活性和促进细胞凋亡。500μmol/L棕榈酸在24 h时细胞活力仅为对照组的54%,cleaved caspase-3蛋白表达则较对照组高3.1倍。棕榈酸诱导p-NF-κB p65在60 min开始升高,120 min时达到高峰增加了2.96倍( P<0.05);IκBα表达水平逐渐下降,以120 min最为明显,下降57%(P<0.05)。10和20μmol/L浓度的NF-κB抑制剂吡咯烷二硫代氨基甲酸盐(PDTC)均能明显抑制棕榈酸诱导的p-NF-κB p65蛋白(1.39±0.12、1.25±0.10比1.76±0.14,均P<0.05);PDTC将cleaved caspase-3和caspase-3蛋白逐渐恢复至接近对照组水平(2.24±0.28比1.29±0.27,0.63±0.01比1.13±0.10,均P<0.05)。结论棕榈酸诱导的MC3T3-E1成骨细胞凋亡可能通过激活NF-κB转导途径起作用。
目的:探討棕櫚痠誘導MC3T3-E1成骨細胞的凋亡是否與覈因子( NF)-κB的激活有關。方法觀察不同濃度(0~1000μmol/L)棕櫚痠孵育MC3T3-E1成骨細胞24 h後對細胞活力和細胞凋亡的影響。細胞活力用四甲基偶氮唑鹽法( MTT法)檢測,細胞凋亡用Hochest 33258染覈和Western印跡法檢測活化型( cleaved ) caspase-3蛋白驗證。用500μmol/L棕櫚痠分彆孵育細胞0~240 min,Western印跡法檢測IκBα、p-NF-κB p65蛋白錶達。結果棕櫚痠呈時間和濃度依賴的方式抑製MC3T3-E1細胞活性和促進細胞凋亡。500μmol/L棕櫚痠在24 h時細胞活力僅為對照組的54%,cleaved caspase-3蛋白錶達則較對照組高3.1倍。棕櫚痠誘導p-NF-κB p65在60 min開始升高,120 min時達到高峰增加瞭2.96倍( P<0.05);IκBα錶達水平逐漸下降,以120 min最為明顯,下降57%(P<0.05)。10和20μmol/L濃度的NF-κB抑製劑吡咯烷二硫代氨基甲痠鹽(PDTC)均能明顯抑製棕櫚痠誘導的p-NF-κB p65蛋白(1.39±0.12、1.25±0.10比1.76±0.14,均P<0.05);PDTC將cleaved caspase-3和caspase-3蛋白逐漸恢複至接近對照組水平(2.24±0.28比1.29±0.27,0.63±0.01比1.13±0.10,均P<0.05)。結論棕櫚痠誘導的MC3T3-E1成骨細胞凋亡可能通過激活NF-κB轉導途徑起作用。
목적:탐토종려산유도MC3T3-E1성골세포적조망시부여핵인자( NF)-κB적격활유관。방법관찰불동농도(0~1000μmol/L)종려산부육MC3T3-E1성골세포24 h후대세포활력화세포조망적영향。세포활력용사갑기우담서염법( MTT법)검측,세포조망용Hochest 33258염핵화Western인적법검측활화형( cleaved ) caspase-3단백험증。용500μmol/L종려산분별부육세포0~240 min,Western인적법검측IκBα、p-NF-κB p65단백표체。결과종려산정시간화농도의뢰적방식억제MC3T3-E1세포활성화촉진세포조망。500μmol/L종려산재24 h시세포활력부위대조조적54%,cleaved caspase-3단백표체칙교대조조고3.1배。종려산유도p-NF-κB p65재60 min개시승고,120 min시체도고봉증가료2.96배( P<0.05);IκBα표체수평축점하강,이120 min최위명현,하강57%(P<0.05)。10화20μmol/L농도적NF-κB억제제필각완이류대안기갑산염(PDTC)균능명현억제종려산유도적p-NF-κB p65단백(1.39±0.12、1.25±0.10비1.76±0.14,균P<0.05);PDTC장cleaved caspase-3화caspase-3단백축점회복지접근대조조수평(2.24±0.28비1.29±0.27,0.63±0.01비1.13±0.10,균P<0.05)。결론종려산유도적MC3T3-E1성골세포조망가능통과격활NF-κB전도도경기작용。
Objective To explore whether palmitate-induced apoptosis of osteoblastic MC 3T3-E1 cell is mediated by an activation of nuclear factor-kappa B ( NF-κB).Methods Cell viability was assessed with methyl thiazolyl tetrazolium ( MTT) assay and cell apoptosis by Hochest 33258 staining.Palmitate was added at different timepoints and dosages.Western blot was used to evaluate the expression levels of IκBα, p-NF-κB p65 and NF-κB p65 protein.Results Palmitate led to a dose-and time-dependent decreases in cell viability and increase in cell apoptosis.Cell viability dropped to 54% and cleaved caspase-3 increased 3.1-fold in cells treated with 500 μmol/L palmitate compared to control.The level of p-NF-κB p65 protein markedly increased at 60 min post-stimulation and reached a 2.96-fold increase of baseline level at 120 min ( P<0.05).The IκBαlevel markedly declined at 60 min post-stimulation and decreased by 57%at 120 min ( P <0.05 ).Compared to the group with palmitate treatment alone , pyrrolidinedithiocarbamic acid (10/20 μmol/L) significantly inhibited the palmitate-induced increase of p-NF-κB p65 ( 1.39 ±0.12, 1.25 ±0.10 vs 1.76 ±0.14, both P<0.05), restored the palmitate-induced decrease of caspase-3(2.24 ± 0.28 vs 1.29 ±0.27, P<0.05)and inhibited the palmitate-induced increase of cleaved caspase-3(0.63 ± 0.01 vs 1.13 ±0.10, P <0.05).Conclusion Palmitate induces apoptosis of MC3T3-E1 cell by an activation of NF-κB.