口腔材料器械杂志
口腔材料器械雜誌
구강재료기계잡지
CHINESE JOURNAL OF DENTAL MATERIALS AND DEVICES
2014年
3期
130-136
,共7页
施琼玲%吴婷婷%李静%汪艳%黄慧
施瓊玲%吳婷婷%李靜%汪豔%黃慧
시경령%오정정%리정%왕염%황혜
微小核糖核酸%成骨分化%骨髓间充质干细胞%肿瘤坏死因子α%骨形态发生蛋白-2
微小覈糖覈痠%成骨分化%骨髓間充質榦細胞%腫瘤壞死因子α%骨形態髮生蛋白-2
미소핵당핵산%성골분화%골수간충질간세포%종류배사인자α%골형태발생단백-2
miRNA%Osteogenic differentiation%BMSCs%TNF-α%BMP-2
目的：探讨肿瘤坏死因子α(TNF-α)对骨形态发生蛋白-2（BMP-2）诱导下小鼠骨髓间充质干细胞(BMSCs)成骨分化的微小核糖核酸（micro ribonucleicacid, miRNA）表达谱的影响。方法给予小鼠BMSCs以下分组刺激：①阴性对照组（ctrl）；②阳性对照组（pos）：BMP-2（200ng/ml）刺激48h；③实验组（48h）：BMP-2（200ng/ml）+TNF-α（10ng/ml）刺激48h；④实验组（72h）：BMP-2（200ng/ml）+TNF-α（10ng/ml）刺激72h，48h和72h后分别提取RNA,应用miRNA芯片检测获得miRNA表达谱，筛选部分差异表达的miRNA进行荧光实时定量PCR（ RT-PCR）验证。结果 miRNA表达谱分析结果表明，与阳性对照组相比，48h双因子刺激下检测到表达上调超过1.5倍的miRNA有44个，72h刺激下检测到22个miRNA，72h与48h相比，检测到24个表达上调超过1.5倍的miRNA，RT-PCR验证与芯片结果基本相符合。结论 miRNA参与调控TNF-α模拟炎症状态下BMP-2诱导BMSCs的成骨分化过程，且相关miRNA在TNF-α作用下表达上调，可能参与调控TNF-α的抑制成骨作用。由此可进一步解释炎性因子对成骨细胞分化的抑制机制，为发现新的药物靶点提供理论依据。
目的：探討腫瘤壞死因子α(TNF-α)對骨形態髮生蛋白-2（BMP-2）誘導下小鼠骨髓間充質榦細胞(BMSCs)成骨分化的微小覈糖覈痠（micro ribonucleicacid, miRNA）錶達譜的影響。方法給予小鼠BMSCs以下分組刺激：①陰性對照組（ctrl）；②暘性對照組（pos）：BMP-2（200ng/ml）刺激48h；③實驗組（48h）：BMP-2（200ng/ml）+TNF-α（10ng/ml）刺激48h；④實驗組（72h）：BMP-2（200ng/ml）+TNF-α（10ng/ml）刺激72h，48h和72h後分彆提取RNA,應用miRNA芯片檢測穫得miRNA錶達譜，篩選部分差異錶達的miRNA進行熒光實時定量PCR（ RT-PCR）驗證。結果 miRNA錶達譜分析結果錶明，與暘性對照組相比，48h雙因子刺激下檢測到錶達上調超過1.5倍的miRNA有44箇，72h刺激下檢測到22箇miRNA，72h與48h相比，檢測到24箇錶達上調超過1.5倍的miRNA，RT-PCR驗證與芯片結果基本相符閤。結論 miRNA參與調控TNF-α模擬炎癥狀態下BMP-2誘導BMSCs的成骨分化過程，且相關miRNA在TNF-α作用下錶達上調，可能參與調控TNF-α的抑製成骨作用。由此可進一步解釋炎性因子對成骨細胞分化的抑製機製，為髮現新的藥物靶點提供理論依據。
목적：탐토종류배사인자α(TNF-α)대골형태발생단백-2（BMP-2）유도하소서골수간충질간세포(BMSCs)성골분화적미소핵당핵산（micro ribonucleicacid, miRNA）표체보적영향。방법급여소서BMSCs이하분조자격：①음성대조조（ctrl）；②양성대조조（pos）：BMP-2（200ng/ml）자격48h；③실험조（48h）：BMP-2（200ng/ml）+TNF-α（10ng/ml）자격48h；④실험조（72h）：BMP-2（200ng/ml）+TNF-α（10ng/ml）자격72h，48h화72h후분별제취RNA,응용miRNA심편검측획득miRNA표체보，사선부분차이표체적miRNA진행형광실시정량PCR（ RT-PCR）험증。결과 miRNA표체보분석결과표명，여양성대조조상비，48h쌍인자자격하검측도표체상조초과1.5배적miRNA유44개，72h자격하검측도22개miRNA，72h여48h상비，검측도24개표체상조초과1.5배적miRNA，RT-PCR험증여심편결과기본상부합。결론 miRNA삼여조공TNF-α모의염증상태하BMP-2유도BMSCs적성골분화과정，차상관miRNA재TNF-α작용하표체상조，가능삼여조공TNF-α적억제성골작용。유차가진일보해석염성인자대성골세포분화적억제궤제，위발현신적약물파점제공이론의거。
Objective In this study, we aimed to investigate the micro ribonucleicacid ( miRNA) expres-sion profile in the tumor necrosis factor (TNF)-α-inhibited osteogenic differentiation of bone marrow mesen-chymal stem cells(BMSCs) introduced by BMP-2. Methods The BMSCs was cultured with/without TNF-αand bone morphogenetic protein(BMP)-2 induction: ①negative control group(ctrl);② positive control(pos):stimulation with BMP-2(200ng/ml)for 48 h;③experimental group(48h):stimulation with BMP-2(200ng/ml) and TNF-α(10ng/ml)for 48 h; ④ experimental group(72h):stimulation with BMP-2(200ng/ml) and TNF-α(10ng/ml)for 72 h. Then the cells were harvested at the indicated times (48h, 72h). miRNA microar-ray technology was used to detect the expression profile of miRNA, and the expression of miRNA was verified by real time quantification-polymerase chain reaction (RT-PCR). Results Microarray analysis found that the expres-sion profile of miRNA changed dramatically. 44 miRNA were upregulated significantly more than 1.5-fold in 48h BMP-2/TNF-αgroup and 22 miRNA in 72h BMP-2/TNF-αgroup, compared with positive control group (P<0.01). Compared with 48h, 24 of up-regulated more than 1.5 times miRNA were detected in 72h BMP-2/TNF-αgroup, which were consistent with the results of miRNA microarray. Conclusion miRNA were involved in the BMP-2-induced osteogenic differentiation process with TNF-αto mimic a state of inflammation, and significantly upregulated miRNA may be involved in the suppression of osteogenesis by TNF-α. These findings can further explain the inhibitory mechanism of inflammatory cytokines on osteoblast differentiation and has important clinical significance.
