CAJ | 학술논문

目的观察地塞米松(DEX)对骨髓间质干细胞(D1细胞系)成脂分化和成骨分化的影响,并探讨抗氧化剂富勒醇的干预作用.方法将D1细胞随机分为:对照组、DEX组、DEX±谷胱甘肽组、DEX±富勒醇组,分组孵育细胞14d后,采用流式细胞仪检测细胞内活性氧簇(ROS)的含量；孵育21d后用油红O染色检测细胞的成脂分化；实时定量聚合酶链反应(Real-time PCR)检测aP2、过氧化物酶体增殖物激活受体γ(PPARγ)、核心结合蛋白因子-2(Runx2)、骨钙素、超氧化物歧化酶(SOD)、过氧化氢酶基因的表达.结果第14天,同对照组比较,经地塞米松处理的细胞中,染色阳性细胞的百分率由91.0％增加到94.7％,染色阳性细胞最低的为经1.0μmmol/L富勒醇处理组.第21天,经DEX处理组的吸光度值为0.439±0.015,较对照组(0.169 ±0.020)及同时应用抗氧剂组均高.最低的为经1.0 μmmol/L富勒醇处理组.DEX明显增加D1细胞中aP2和PPARγ基因的表达,降低Runx2、骨钙素、SOD和过氧化氢酶基因的表达,增加ROS水平,促进成脂分化,富勒醇可明显拮抗DEX诱导的上述效应.结论富勒醇可能通过降低氧化应激水平,进而减少成脂分化、增加成骨分化.
목적 관찰지새미송(DEX)대골수간질간세포(D1세포계)성지분화화성골분화적영향,병탐토항양화제부륵순적간예작용.방법 장D1세포수궤분위:대조조、DEX조、DEX±곡광감태조、DEX±부륵순조,분조부육세포14d후,채용류식세포의검측세포내활성양족(ROS)적함량；부육21d후용유홍O염색검측세포적성지분화；실시정량취합매련반응(Real-time PCR)검측aP2、과양화물매체증식물격활수체γ(PPARγ)、핵심결합단백인자-2(Runx2)、골개소、초양화물기화매(SOD)、과양화경매기인적표체.결과 제14천,동대조조비교,경지새미송처리적세포중,염색양성세포적백분솔유91.0％증가도94.7％,염색양성세포최저적위경1.0μmmol/L부륵순처리조.제21천,경DEX처리조적흡광도치위0.439±0.015,교대조조(0.169 ±0.020)급동시응용항양제조균고.최저적위경1.0 μmmol/L부륵순처리조.DEX명현증가D1세포중aP2화PPARγ기인적표체,강저Runx2、골개소、SOD화과양화경매기인적표체,증가ROS수평,촉진성지분화,부륵순가명현길항DEX유도적상술효응.결론 부륵순가능통과강저양화응격수평,진이감소성지분화、증가성골분화.
Objective To evaluate the effect of fullerol,a powerful antioxidant,on adipogenic and osteogenic differentiation of a mouse bone marrow-derived multipotent cell line,D1.Methods D1 cells were divided into five groups:D1 cells; D1 cells treated with 1 x 10-5 mol/L dexamethasone (DEX) ; (3) D1 cells treated with 1 x 10-5 mol/L DEX and 1 x 10-5 mol/L glutathione (GSH) ; (4) D1 cells treated with1 ×10-5 mol/L DEX and 1 ×10-7 mol/L fullerol; (5) Dl cells treated with 1 ×10-7 mol/L DEX and 1 x10-6 mol/L fullerol.Reactive oxygen species (ROS) were measured by staining the cells at day 21 with a fluorescent probe,2',7'-dichlorodihydrofluorescin diacetate (DCFH-DA).Lipid droplets within cells were assessed by staining with Oil Red O at day 21.The gene expression was detected by using quantitative realtime quantitative polymerase chain reaction (Real-time PC R).Results The 14 d,compared with the control group (91.0％),the percentage of positive cells was 94.7％ in dexamethasone-treated cells,the lowest positive cells was treated by 1.0 μmmol/L fullerol.The 21 d,the highest optical density value of DEX-treated group was 0.439 ±0.015,compared with the control group (0.169 ±0.020) and the other which treated with antioxidant.The lowest was treated by 1.0 μmmol/L fullerol.DEX increased the expression of aP2 andperoxisome proliferator activated receptor γ (PPARγ) gene significantly,reduced the expression of related transcription factor-2 (Runx2),osteocalcin,superoxide dismutase (SOD) and catalase gene,increased the levels of ROS,and promoted adipogenic differentiation,which could be antagonized significantly by fullerenol.Conclusion Fullerol antagonizes dexamethasone-induced oxidative stress and adipogenesis by reducing the leveal of ROS.