N-Ac-L-Leu-PEI/微小RNA-34a复合物对大鼠颅骨缺损成骨的影响
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摘要
目的:将N-Ac-L-Leu-PEI/微小RNA(microRNA,miR)-34a复合物用于大鼠颅骨缺损模型,探讨miR-34a对骨缺损区成骨的影响。方法:Wistar雄性大鼠32只,制备标准大鼠颅骨缺损模型,随机分为4组,分别于缺损区覆盖载有磷酸盐缓冲液(phosphate buffered salin,PBS)、N-Ac-L-Leu-PEI、N-Ac-L-LeuPEI/miR-34a复合物、N-Ac-L-Leu-PEI/miR-34aNC复合物的口腔生物膜,饲养10周后处死大鼠,通过锥形束CT(cone beam computerized tomography,CBCT)、微型CT(microcomputed tomography,micro-CT)、反转录-聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)等方法检测大鼠颅骨缺损区新骨形成情况。结果:N-Ac-L-Leu-PEI/miR-34a复合物组缺损区骨新生最为明显,新生骨量与总组织量之比(bone volume/total volum,BV/TV)、骨小梁厚度(trabecular thickness,Tb.Th)值均高于其它组,成骨标志性基因Runt相关转录因子2(runt-related transcription factor 2,Runx2)、Osterix转录因子(serine protease 7,SP7)及Ⅰ型胶原(CollagenⅠ,ColⅠ)的mRNA表达显著上调。结论:对实验性大鼠颅骨缺损模型,N-Ac-LLeu-PEI/miR-34a具有促进缺损区成骨的作用。
Objective:To study the effects of N-Ac-L-Leu-PEI/micro RNA(miR)-34 acomplex on osteogenisis of rat cranial defects.Methods:Thirty-two Wistar male rats were randomly divided into four groups and standard cranial defects were prepared on each rat.Biofilms loaded with PBS,N-Ac-L-Leu-PEI,N-Ac-LLeu-PEI/miR-34 acomplex,and N-Ac-L-Leu-PEI/miR-34 aNC complex covered the defect.After ten weeks culture,the rats were sacrificed and the development of osteogenisis was detected by CBCT,micro-CT and RT-PCR.Results:Bone formation was evident in the defects of N-Ac-L-Leu-PEI/miR-34 acomplex group.The BV/TV values and Tb.Th values of N-Ac-L-Leu-PEI/miR-34 acomplex group increased significantly,suggesting new bone formation.The expression of Runx2,SP7,and Col I mRNA was upregulated by N-Ac-L-Leu-PEI/miR-34 acomplex,which were specific factors of osteogenisis.Conclusion:N-Ac-L-Leu-PEI/miR-34 acomplex can promote osteogenesis in rat cranial defects.
Objective:To study the effects of N-Ac-L-Leu-PEI/micro RNA(miR)-34 acomplex on osteogenisis of rat cranial defects.Methods:Thirty-two Wistar male rats were randomly divided into four groups and standard cranial defects were prepared on each rat.Biofilms loaded with PBS,N-Ac-L-Leu-PEI,N-Ac-LLeu-PEI/miR-34 acomplex,and N-Ac-L-Leu-PEI/miR-34 aNC complex covered the defect.After ten weeks culture,the rats were sacrificed and the development of osteogenisis was detected by CBCT,micro-CT and RT-PCR.Results:Bone formation was evident in the defects of N-Ac-L-Leu-PEI/miR-34 acomplex group.The BV/TV values and Tb.Th values of N-Ac-L-Leu-PEI/miR-34 acomplex group increased significantly,suggesting new bone formation.The expression of Runx2,SP7,and Col I mRNA was upregulated by N-Ac-L-Leu-PEI/miR-34 acomplex,which were specific factors of osteogenisis.Conclusion:N-Ac-L-Leu-PEI/miR-34 acomplex can promote osteogenesis in rat cranial defects.
引文
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[3]Chen L,Holmstrφm K,Qiu W,et al.MicroRNA-34ainhibits osteoblast differentiation and in vivo bone formation of human stromal stem cells[J].Stem Cells,2014,32(4)∶902-912
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[6]丁小博.以PEI衍生物为载体构建肿瘤基因递送体系:从小分子到大分子的修饰[D].吉林大学,2015
[7]Li Z,Zhang L,Li Q.Induction of apoptosis in cancer cells through N-acetyl-l-leucine-modified polyethyleniminemediated p53gene delivery[J].Colloids Surf B Biointerfaces,2015,135∶630-638
[8]Fan C,Jia L,Zheng Y,et al.MiR-34apromotes osteogenic differentiation of human adipose-derived stem cells via the RBP2/NOTCH1/CYCLIN D1 coregulatory network[J].Stem Cell Reports,2016,7(2)∶236-248
[9]Park H,Park H,Pak HJ,et al.miR-34ainhibits differentiation of human adipose tissue-derived stem cells by regulating cell cycle and senescence induction[J].Differentiation,2015,90(4-5)∶91-100
[10]Li XJ,Ren ZJ,Tang JH.MicroRNA-34a:apotential therapeutic target in human cancer[J].Cell Death Dis,2014,5(7)∶e1327
[11]李晓光,王一珠,郭斌.慢性牙周炎中肿瘤坏死因子α对骨髓间充质干细胞成骨分化的调控作用[J].华西口腔医学杂志,2017,35(3)∶334-338
[12]赵娜,方慧,唐亚平,等.MAPK信号转导通路与慢性牙周炎的相关研究进展[J].口腔医学研究,2017,33(9)∶1012-1015
[13]Zha X,Sun B,Zhang R,et al.Regulatory effect of MicroRNA-34aon osteogenesis and angiogenesis in glucocorticoid-induced osteonecrosis of the femoral head[J].J Orthop Res,2018,36(1)∶417-424
[14]Adhami MD,Rashid H,Chen H,et al.Loss of Runx2in committed osteoblasts impairs postnatal skeletogenesis[J].J Bone Miner Res,2015,30(1)∶71-82
[15]刘引,申玉芹,高涵,等.MT01对牙龈卟啉单胞菌感染成骨细胞MG63中Ⅰ型胶原mRNA表达的促进作用及其意义[J].吉林大学学报(医学版),2016,42(4)∶681-684