Expression of activating transcription factor 4 in osteogenic induction of bone marrow mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.2014.01.004

Abstract

Abstract:

BACKGROUND: Activating transcription factor 4 is found as an activating factor that can regulate osteogenic differentiation and function, and plays a critical role in the osteogenic differentiation.
OBJECTIVE: To investigate the expression and significance of activating transcription factor 4 in the osteogenic differentiation of bone marrow mesenchymal stem cells from Sprague-Dawley rats.
METHODS: Bone marrow mesenchymal stem cells from Sprague-Dawley rats were cultured using the whole bone marrow adherence method, and ossification revulsant was added to induce passage 3 cells. Cells with no osteogenic induction served as controls. RT-PCR and western blot assay were employed to dynamically monitor expression of activating transcription factor 4.
RESULTS AND CONCLUSION: RT-PCR results showed that activating transcription factor 4 mRNA expression increased with the increasing osteogenic differentiation, and peaked at day 16. Western blot analysis showed that the protein expression of activating transcription factor 4 tended to increase with the increasing osteogenic differentiation, peaked at day 16 and still maintained at a higher level at day 19. Compared with the uninduced cells, activating transcription factor 4 in the induced cells exhibited a higher expression at mRNA and protein levels (P < 0.05). These findings indicate that activating transcription factor 4 expression is elevated during osteogenic differentiation, showing a positive correlation with osteogenic ability of bone marrow mesenchymal stem cells.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: stem cells, mesenchymal stem cells, osteoblasts, activating transcription factor 4

CLC Number:

R394.2

Wu Dan, Li Su-ling, Wang Lu, Xu Ling, Xiang Hua-ju. Expression of activating transcription factor 4 in osteogenic induction of bone marrow mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(1): 21-26.

Figures/Tables 4

References

[1] Yang X,Matsuda K,Bialek P,et al.ATF4 is a substrate of RSK2 and an essential regulator of osteoblast biology:Implication for Cofin-Lowry syndrome.Cell.2004; 17:387-398.

[2] Mendes SC,Tibbe JM,Veenhof M,et al.Bone tissue-engineered implants using human bone nlarrow stromal cells:effect of culture conditions and donor age.Tissue Eng.2002;(8):911-920.

[3] 程雷,聂林,谢青,等.大鼠骨髓基质干细胞的培养鉴定及向成骨细胞诱导分化的实验研究[J].中华物理医学与康复杂志,2005, 27(3): 138-140.

[4] 张琪,陈规划.诱导多能肝细胞研究进展-细胞的炼丹术[J].器官移植,2010,1(1):52-54.

[5] 朱勇,陈良万,林若柏,等.SD大鼠骨髓间充质干细胞体外培养、表型鉴定及标记[J].吉林大学学报:医学版,2009,35(2):326-329.

[6] 杨柳,段小军.骨组织工程学中种子细胞研究的前沿问题[J].中国临床康复,2002,6(4):532-533.

[7] De Ugarte DA, Alfonso Z, Zuk PA, et al.Diferential expression of stem cell mobilization••associated molecules on multi-lineage ce lls from adipose tissue and bone mSlTOW. Immunoln.2003;89(23):267-270.

[8] 孙晓雷.牛脱细胞骨基质生物力学及其黏附性的实验研究[D].天津:天津医科大学,2007.

[9] 王广军,王爱萍,郑修启,等.骨髓间充质干细胞与基因治疗[J].泰山医学院学报,2006,27(2):179-181.

[10] 阳玉群,莫雪安,农伟东,等.全骨髓贴壁改良培养法体外分离培养大鼠骨髓间充质干细胞[J].广西医科大学学报,2013,30(2): 171-174.

[11] 程庆,邓湘东,曹卉,等. 改良贴壁法培养大鼠骨髓间充质干细胞[J]. 中国热带医学,2013,13(8):931-934.

[12] 银广悦,陈素萍,丁俊丽,等.小鼠骨髓间充质干细胞的体外分离培养和鉴定方法学探讨[J].中国实验诊断学,2013,17(4): 647-650.

[13] 肖仕辉,韦庆军,赵劲民,等.全骨髓贴壁法培养兔骨髓间充质干细胞体外定向成骨诱导分化及鉴定[J].中国组织工程研究,2013, 17(6):1069-1074.

[14] 戚宗泽,罗云飞,侯勇,等.兔骨髓间充质干细胞的分离、体外培养、鉴定及成脂诱导[J].昆明医科大学学报,2013,34(1):28-31.

[15] 吴玉琼,江凌勇,张鹏,等.骨质疏松大鼠骨髓间充质干细胞的培养方法及生物学特性鉴定[J].中国口腔颌面外科杂志,2013,11(1): 8-14.

[16] 何丁文,殷嫦嫦,殷明.全骨髓贴壁法分离培养大鼠骨髓间充质干细胞研究[J].九江学院学报:自然科学版,2012,(4):67-70.

[17] 何丁文,邬亚华,殷明,等.不同胎牛血清、培养瓶体外培养大鼠骨髓间充质干细胞效果比较[J].山东医药,2012,52(38): 30-32.

[18] 韩伟,李争艳,刘杨,等. Wistar大鼠骨髓间充质干细胞的原代培养和鉴定[J].哈尔滨医科大学学报,2011,45(2):135-137.

[19] 张守信,刘晓妍,华秀峰,等.新生小鼠心肌细胞的纯化培养与鉴定[J].中国医药指南,2012,10(26): 20-22.

[20] 刘贤华,仝青英,白晓东.人骨髓间充质干细胞体外不同分离与培养方法的比较[J].武警医学,2012,23(10):836-839.

[21] 曾瑞霞,单伟,房艳,等.密度梯度离心法结合贴壁法体外分离培养大鼠骨髓间充质干细胞[J].解剖科学进展,2012,18(5): 438-441.

[22] 张君红,姜海行,覃山羽,等.全骨髓细胞贴壁法分离与培养大鼠骨髓间充质干细胞的多向分化能力[J].中国组织工程研究,2012, 16(36):6685-6689.

[23] 盛瑾,许官学,石蓓,等.全骨髓法培养SD大鼠骨髓间充质干细胞及其生物学特性[J].遵义医学院学报,2011,34(2):132-136.

[24] 张利铭,陈智超,邹萍,等.全骨髓贴壁法分离培养人骨髓间充质干细胞及成骨诱导分化[J].临床血液学杂志,2012,25(3):317-319.

[25] 许峰,李宝平,张雷,等.全骨髓贴壁改良法分离培养人骨髓间充质干细胞及标记鉴定[J].中国医学工程,2012,20(2):63-63.

[26] 李晓峰,赵劲民,苏伟,等.大鼠骨髓间充质干细胞的培养与鉴定[J]. 中国组织工程研究与临床康复,2011,15(10):1721-1724.

[27] 谭琦,黄政德,王立凤.大鼠骨髓间充质干细胞分离和培养方法的研究[J].湖南中医药大学学报,2011,31(7): 74-76.

[28] 李树法,张敏.贴壁和缩短胰酶消化时间培养大鼠骨髓间充质干细胞[J].中国组织工程研究与临床康复,2011,15(36): 6687- 6690.

[29] 赵林,冯智慧,焦淑贤,等.全骨髓贴壁法分离培养大鼠骨髓间充质干细胞及其生物学特性[J].中国组织工程研究与临床康复,2011, 15(32):5923-5927.

[30] 宗英,袁伯俊,陆国才.活化转录因子4在辐射诱导的损伤中的作用及机制[J].中国药理学与毒理学杂志,2013,27(3):172.

[31] 柴路维,姜蓉,何小茜,等. ATF4基因在小鼠胚胎围着床期子宫内膜的表达[J].基础医学与临床,2012,32(11):1293-1297.

[32] 李发祥,杨荣,陈婷芳,等.斑马鱼基因ATF4多克隆抗体的制备[J]. 激光生物学报,2012,21(1):56-59.

[33] 陈超,吴望军,熊远著. 猪ATF4基因多态性与生产性状的关联及基因表达分析[J].遗传,2011,33(12):1347-1352.

[34] 潘红,王军,朱冠南,等.缺氧缺血性脑损伤新生大鼠磷酸化真核翻译起始因子2α和激活转录因子4表达的变化[J]. 南京医科大学学报:自然科学版,2011,31(11):1573-1575.

[35] 刘晓燕,戴爱国,谭双香.慢性阻塞性肺疾病中转录因子ATF3/ ATF4与Nrf2的表达及相互作用[J].中国病理生理杂志,2011, 27(10):1961.

[36] 刘晓燕,戴爱国,李洁.转录因子ATF3和ATF4在大鼠慢性阻塞性肺疾病中的表达及作用[J].中国呼吸与危重监护杂志,2011, 10(2):121-125.

[37] 王万东,陈东风.非酒精性脂肪变性肝细胞模型中ATF4基因的表达及其意义[J].第三军医大学学报,2010,32(14): 1487-1490.

[38] 申向民,杨期东,谭利明,等.大鼠局灶性脑缺血再灌后转录因子ATF4的表达变化[J].中风与神经疾病杂志,2010,27(4):330-332.

[39] 王春玲,魏福兰,赵艳红,等.离心力作用下人牙周膜细胞转录活化因子4的表达[J].上海口腔医学,2007,16(4):422-426.

[40] Yang X,Karsenty G.ATF4,the osteoblast accumulation of whichis determ ined post-translationally,can induce 0ste0blastspeci6cgene expression in non-osteoblastic Cells. Biol Chem.2004;279:47109-47114.

[41] 庄淑波,刘毅,陈克明,等.大鼠骨髓间充质干细胞体外分离、纯化与培养适宜条件的筛选[J].中国组织工程研究与临床康复, 2007, 11(20):3886-3991.

[42] 魏福兰.ATF4与正畸牙周组织改建相关的实验研究[D].济南:山东大学,2007.

[43] Block MA, Kent JN.Placement of endosseous implants into tooth extractions sites. Oral Maxillofacial Surg.2010;(9): 269-276.

[44] 吴丹.即刻种植即刻加载对骨结合界面形成的影响[D].重庆:重庆医科大学,2012.

[45] Nail GA, Stein S, Korhi M, et al. Evaluation of endosseous implants placed in fresh extraction sites in dogs. Dent Res. 2010(6):347-351.