MicroRNA-124a对SCI继发性损伤的影响及相关机制研究Effects of MicroRNA-124a on SCI Secondary Injury and the Underlying Mechanism
张芬
摘要(Abstract):
探究microRNA-124a(miR-124a)对脊髓损伤(spinal cord injury,SCI)继发性损伤的影响及相关机制.取90只健康成年Wistar大鼠建立SCI模型,随机分为3组,第1组术后仅注射生理盐水,为未治疗组;第2组术后注射骨髓基质细胞源神经干细胞(BMSCs-D-NSCs)治疗,为正常移植组;第3组术后注射携带miR-124a的骨髓基质细胞源神经干细胞(miR-124a-BMSCs-D-NSCs),为转染移植组.另选取60只健康成年Wistar大鼠作为对照组,仅给予假手术处理.采用HE染色观察大鼠的组织形态学变化,采用实时荧光定量PCR技术检测大鼠的miR-124a表达情况,并对损伤灶进行显微测量计算,比较脊髓功能恢复的BBB评分情况及脊髓组织损伤大小的情况.结果显示,SCI后,脊髓组织中miR-124a基因表达显著降低,脊髓神经功能也明显降低,miR-124a-BMSCs-D-NSCs能够有效地改善SCI,减少脊髓损伤组织范围及体积,还能改善中枢神经的功能.
关键词(KeyWords): microRNA-124a;脊髓损伤;神经功能;作用机制
基金项目(Foundation):
作者(Author): 张芬
参考文献(References):
- [1]LEE B B,CRIPPS R A,FITZHARRIS M,et al.The global map for traumatic spinal cord injury epidemiolopy:update 2011,global incidence rate[J].Spinal cord,2014,52(2):110—115.
- [2]STRICKLAND E R,HOOK M A,BALARAMAR S,et al.MicroRNA dysregulation following spinal cord contusion:implications for neural plasticity and repair[J].Neuroscience,2011,186(186):146—160.
- [3]KRICHEVSKY A M.MicroRNA profiling:from dark matter to white matter,or identifying new players in neurobiology[J].Scientific world journal,2007,7(S2):155—166.
- [4]KOSIK K S.The neuronal microRNA system[J].Nature reviews neuroscience,2006,7(12):911—920.
- [5]SHAFI G,ALIYA N,MUNSHI A.MicroRNA signatures in neurological disorders[J].The Canadian journal of neurological sciences Le journal Canadien des sciences neurologiques,2010,37(2):177—185.
- [6]BAREYRE F M,SCHWAB M E.Inflammation,degeneration and regeneration in the injured spinal cord:insights from DNA microarrays[J].Trends in neurosciences,2003,26(10):555—563.
- [7]di GIOVANNI S,KNOBLACH S M,BRANDOL C,et al.Gene profiling in spinal cord injury shows role of cell cycle in neuronal death[J].Annals of neurology,2003,53(4):454—468.
- [8]NESIC O,SVRAKIC N M,XU G Y,et al.DNA microarray analysis of the contused spinal cord:effect of NMDA receptor inhibition[J].Journal of neuroscience research,2002,68(4):406—423.
- [9]NIETO-DIAZ M,ESTEBAN F J,REIGADA D,et al.MicroRNA dysregulation in spinal cord injury:causes,consequences and therapeutics[J].Frontiers in cellular neuroscience,2014,8(9):53.
- [10]张雨禄,胡三元,等.乳腺癌转移抑制因子(BRMS1)在人乳腺癌细胞系中的表达受体研究[J].郑州大学学报(理学版),2014,46(1):94—97.
- [11]BAK M,SILAHTAROGLU A,MOLLER M,et al.MicroRNA expression in the adult mouse central nervous system[J].RNA,2008,14(3):432—444.
- [12]HOHJOH H,FUKUSHIMA T.Expression profile analysis of microRNA(miRNA)in mouse central nervous system using a new miRNA detection system that examines hybridization signals at every step of washing[J].Gene,2007,391(s1/s2):39—44.
- [13]FARH K K,GRIMSON A,JAN C,et al.The widespread impact of mammalian microRNAs on mRNA repression and evolution[J].Science,2005,310(5755):1817—1821.
- [14]LANDGRAF P,RUSU M,SHERIDAN R,et al.A mammalian microRNA expression atlas based on small RNA library sequencing[J].Cell,2007,129(7):1401—1414.
- [15]YUNTA M,NIETO-DIAZ M,ESTEBAN F J,et al.MicroRNA dysregulation in the spinal cord following traumatic injury[J].PLo S one,2012,7(4):120—123.
- [16]NAKANISHI K,NAKASA T,TANAKA N,et al.Responses of microRNAs 124a and 223 following spinal cord injury in mice[J].Spinal cord,2010,48(3):192—196.
- [17]HUANG Y,JIA X,BAI K,et al.Effect of fluid shear stress on cardiomyogenic differentiation of rat bone marrow mesenchymal stem cells[J].J Arch Med Res,2010,41(7):497—505.
- [18]张春燕,张自强,刘玉梅,等.大鼠雪旺细胞体外分离培养纯化及鉴定[J].河南科技大学学报(自然科学版),2015,36(4):82—84.
- [19]MARL P,HINDOCHA S,MARL R,et al.Adult mesenchymal stem cells and cell surface characterization:a systematic review of the literature[J].J Open Orthop J,2011,5(suppl2):253—260.
- [20]BARNAB G F,SCHWINDT T T,CALCAGNOTTO M E,et al.Chemically-induced rat mesenchymal stem cells adopt molecular properties of neuronal.like cells but do not have basic neuronal functional properties[J].PLo S one,2009,4(4):e5222.
- [21]RADTKE C,SCHMITZ B,SPIES M,et al.Peripheral glial cell differentiation from neurospheres derived from adipose mesenchymal stem cells[J].Int J Devl Neuroscience,2009,27(8):817—823.
- [22]程腾,李佳佳,贺小英,等.培养方法及胎牛血清浓度对人胎儿成纤维细胞体外生长的影响[J].信阳师范学院学报(自然科学版),2013,26(4):523—526.
- [23]WISLET-GENDEBIEN S,LAUDET E,NEIRINCKX V,et al.Adult bone marrow:which stem cells for cellular therapy protocols in neurodegenerative disorders?[J].J biomed biotechnol,2012,2012(1):111—113.
- [24]MIN K J,JIN S J,JEE C,et al.MicroRNA 486 is a potentially novel target for the treatment of spinal cord injury[J].Brain,2012,135(4):1237—1252.
- [25]李佳佳,程腾,贺小英,等.二甲基亚砜和胎牛血清对293T细胞冷冻效果的影响[J].信阳师范学院学报(自然科学版),2013,26(2):217—220.
- [26]BASSO D M,BEATTIE M S,BRESNAHAN J C.Graded histological and motor outcomes after spinal cord contusion using the NYU weight.drop device versus transaction[J].Exp Neur,1996,139(6):244—256.
- [27]CHOPP M,ZHANG X H,LI Y,et al.Spinal cord injury in rat:treatment with bone marrow stromal cell transplantation[J].Neuroreport,2000,11(13):3001—3005.
- [28]BERNSTEIN E,KIM S Y,CARMELL M A,et al.Dicer is essential for mouse development[J].Nature genetics,2003,35(3):215—217.
- [29]CUELLAR T L,DAVIS T H,NELSON P T,et al.Dicer loss in striatal neurons produces behavioral and neuroanatomical phenotypes in the absence of neurodegeneration[J].Proceedings of the national academy of sciences of the United States of America,2008,105(14):5614—5619.
- [30]ZEHIR A,HUA L L,MASKA E L,et al.Dicer is required for survival of differentiating neural crest cells[J].Developmental biology,2010,340(12):459—467.
- [31]KAWASE-KOGA Y,OTAEGI G,SUN T.Different timings of Dicer deletion affect neurogenesis and gliogenesis in the developing mouse central nervous system[J].Developmental dynamics:an official publication of the American association of anatomists,2009,238(11):2800—2812.
- [32]FIORE R,SIEGEL G,SCHRATT G.MicroRNA function in neuronal development,plasticity and disease[J].Biochimica et biophysica acta,2008,1779(8):471—478.
- [33]BARTEL D P.MicroRNAs:genomics,biogenesis,mechanism,and function[J].Cell,2004,116(2):281—297.
- [34]CHENG X,HE D,DONG X D,et al.miR-124a is epigenetically regulated and acts as a tumor suppressor by controlling multiple targets in uveal melanoma[J].Investigative ophthalmology visual science,2013,54(3):2248—2256.