Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (17): 3688-3694.doi: 10.3864/j.issn.0578-1752.2013.17.018

• ANIMAL SCIENCE·VETERINARY SCIENCERE·SOURCE INSECT • Previous Articles     Next Articles

Effect of h1-Calponin Gene on in Vitro Osteogenesis Differentiation of Bone Marrow Mesenchymal Stem Cells of Wuzhishan Minipig

 HE  Wei, WU  Tian-Wen, HUANG  Lei, ZHAO  Wei-Min, XIA  Ying, TANG  Fang, MOU  Yu-Lian, LI  Kui   

  1. Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193
  • Received:2012-02-04 Online:2013-09-01 Published:2013-04-23

PDF

505

Cited

3

Abstract: 【Objective】 The objective of the present study is to explore the relationship between h1-calponin gene and osteoblast differentiation of Wuzhishan minipig’s bone marrow mesenchymal stem cells (BM MSCs) and further understand the mechanism of osteoblast differentiation.【Method】BM MSCs were induced to osteoblast differentiation by adding osteogenesis differentiation medium. Alizarin Red staining was used to indentify the degree of osteoblast differentiation of BM MSCs. The expression profile of h1-calponin gene and the osteo-related genes osteopontin (OPN), osteocalcin (OCN), core binding factor α1 (Cbfα1) and core binding factor β (Cbfβ) were analyzed by quantitative PCR.【Result】Using Alizarin Red staining, it was found that the nodules of cells were increasing as the induction time went on. Q-PCR results showed that the expression patterns of OPN and OCN (two marker genes of osteogesis) were the same. Both OPN and OCN genes increased from day 0 to day 14, peaked at day 14 and finally declined. The osteo-related genes, Cbfα1 and Cbfβ, were highestly expressed at day 7 and then decreased gradually, but still higher than pre-induction. The expression of h1-calponin gene was gradually decreased during the induction and was nearly undetected from day 14 to 21.【Conclusion】Results of the experiment suggested there is a negative correlation between the expression of h1-calponin gene and osteoblast differentiation of Wuzhishan minipig’s BM MSCs is in vitro. The h1-calponin gene may be a negative-regulated gene during osteoblast differentiation.

Key words: h1-calponin , BM MSCs , osteoblast differentiation , negative regulation

[1]张向荣, 刘德伍, 郭光华. 骨髓间充质干细胞生物学特性及其在组织修复中的应用. 中国组织工程研究与临床康复, 2008, 12(8): 1513-1516.

Zhang X R, Liu D W, Guo G H. Biological characteristics of mesenchymal stem cells and its application in tissue repair. Journal of Clinical Rehabilitative Tissue Engineering Research, 2008, 12(8): 1513-1516. (in Chinese)

[2]Liu G L, Lu Y F, Li W J, Xiao H Z, Sun G G, Yu F, Xiang X X, Zhang H Q, Liu X L, Shi Y P, Li S. Differentiation of marrow-derived islet-like cells and their effects on diabetic rats. Chinese Medical Journal, 2010, 123(22): 3347-3350.

[3]Wang J C, Xia L, Song X B, Wang C E, Wei F C. Transplantation of hypoxia preconditioned bone marrow mesenchymal stem cells improves survival of ultra-long random. Chinese Medical Journal, 2011, 124(16): 2507-2511.

[4]Chang Q, Yan L, Wang C Z, Zhang W H, Hu Y Z, Wu B Y. In vivo transplantation of bone marrow mesenchymal stem cells accelerates repair of injured gastric mucosa in rats. Chinese Medical Journal, 2012, 125(6): 1169-1174.

[5]Friedenstein A J, Piatetzky S II, Petrakova K V. Osteogenesis in transplants of bone marrow cells. Journal of Embryology and Experimental Morphology, 1966, 16(3): 381-390.

[6]Rossignol J, Boyer C, Thinard R, Remy S, Dugast A S, Dubayle D, Dey N D, Boeffard F, Delecrin J, Heymann D, Vanhove B, Anegon I, Naveilhan P, Dunbar G L, Lescaudron L. Mesenchymal stem cells induce a weak immune response in the rat striatum after allo or xenotransplantation. Journal of Cellular and Molecular Medicine, 2009, 13(8): 2547-2558.

[7]Shi Y F, Su J J, Roberts A I, Shou P, Rabson A B, Ren G W. How mesenchymal stem cells interact with tissue immune responses. Trends in Immunology, 2012, 33(3): 136-143.

[8]Yan Z Q, Hang D H, Guo C G, Chen Z R. Fate of mesenchymal stem cells transplanted to osteonecrosis of femoral head. Journal of Clinical Orthopaedics, 2009, 27(4): 442-446.

[9]Wang B L, Sun W, Shi Z C, Zhang N F, Yue D B, Guo W S, Xu S Q, Lou J N, Li Z R. Treatment of nontraumatic osteonecrosis of the femoral head with the implantation of core decompression and concentrated autologous bone marrow containing mononuclear cells. Archives of Orthopaedic and Trauma Surgery, 2010, 130(7): 859-865.

[10]季卫锋, 丁伟航, 马镇川, 厉驹, 童培建. 三通道髓芯钻孔减压加DBM、自体骨髓干细胞治疗早期股骨头坏死. 中国骨伤, 2008, 21(10): 776-778.

Ji W F, Ding W H, Ma Z C, Li J, Tong P J. Three-tunnels core decompression with implantation of bone marrow stromal cells(bMSCs) and decalcified bone matrix (DBM) for the treatment of early femoral head necrosis. China Journal of Orthopaedics and Traumatology, 2008, 21(10): 776-778. (in Chinese)

[11]Kadiyala S, Young R G, Thiede M A, Bruder S P. Culture expanded canine mesenchymal stem cells possess osteochondrogenic potential in vivo and in vitro. Cell Transplant, 1997, 6(2): 125-134.

[12]Yoshikawa H，Taniguchi S I, Yamamura H, Mori S, Sugimoto M, Miyado K, Nakamura K, Nakao K, Katsuki M, Shibata N, Takahashi K. Mice lacking smooth muscle calponin display increased bone formation that is associated with enhancement of bone morphogenetic protein responses. Genes to Cells, l998, 3: 685-695.

[13]Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods, 2001, 25(4): 402-408.

[14]Zhao Y M, Waldman S D, Flynn L E. The effect of serial passaging on the proliferation and differentiation of bovine adipose-derived stem cells. Cells Tissues Organs, 2012, 195(5): 414-427.

[15]Jackson W M, Nesti L J, Tuan R S. Potential therapeutic applications of muscle-derived mesenchymal stem and progenitor cells. Expert Opinion on Biological Therapy, 2010, 10(4): 505-517.

[16]Tobiasch E. Adult human msenchymal stem cells as source for future tissue engineering. Forschungsspitzen and Spitzenforschung, 2009: 329-338.

[17]Bruno S, Bussolati B, Grange C, Collino F, Cantogno L V, Herrera M B, Biancone L, Tetta C, Segoloni G, Camussi G. Isolation and characterization of resident mesenchymal stem cells in human glomeruli. Stem Cells and Development, 2009, 18(6): 867-880.

[18]Secco M, Zucconi E, Vieira N M, Fogaca L L Q¸ Cerqueira A, Carvalho M D F, Jazedje T, Okamoto O K, Muotri A R, Zatz M. Multipotent stem cells from umbilical cord: cord is richer than blood! Stem Cells, 2008, 26: 146-150.

[19]Zhao J J, Chen N Y, Shen N, Zhao H, Wang D L, Shi J, Wang Y, Cui X F, Yan Z Y, Xue H. Transplantation of human umbilical cord blood mesenchymal stem cells to treat a rat model of raumatic brain injury. Neural Regeneration Research, 2012, 7(10): 741-748.

[20]Lee J M, Jung J, Lee H J, Jeong S J, Cho K J, Hwang S G, Kim G J. Comparison of immunomodulatory effects of placenta mesenchymal stem cells with bone marrow and adipose mesenchymal stem cells. International Immunopharmacology, 2012, 13(2): 219-224.

[21]Zheng H, Guo Z, Ma Q, Jia H, Dang G. Cbfa1/ osf2 transduced bone marrow stromal cells facilitate bone formation in vitro and in vivo. Calcified Tissue International, 2004, 74: 194-203.

[22]Komori T. Runx2, a multifunctional transcription factor in skeletal development. Journal of Cellular Biochemistry, 2002, 87(1): 1-8.

[23]Xuan D Y, Sun X, Yan Y X, Xie B Y, Xu P P, Zhang J C. Effect of cleidocranial dysplasia-related novel mutation of RUNX2 on characteristics of dental pulp cells and tooth. Journal of Cellular Biochemistry, 2010, 111: 1473-1481.

[24]Michael Cohen Jr M. Perspectives on RUNX genes: an update. American Journal of Medical Genetics, 2009, 149A: 2629-2646.

[25]Lu Y B, Li Y C, Cavender A C, Wang S Z, Mansukhani A, D’Souza1 R N. Molecular studies on the roles of Runx2 and Twist1 in regulating FGF signaling. Developmental Dynamics, 2012, 241: 1708-1715.

[26]Lien C Y, Lee O K, Su Y. Cbfb enhances the osteogenic differentiation of both human and mouse mesenchymal stem cells induced by Cbfa‐1 via reducing its ubiquitination‐mediated degradation. Stem Cells, 2007, 25(6): 1462-1468.

[27]Tran T C, Kimura K, Nagano M, Yamashita T, Ohneda K, Sugimori H, Sato F, Sakakibara Y, Hamada H, Yoshikawa H, Hoang S N, Ohneda O. Identification of human placenta-derived mesenchymal stem cells involved in re-endothelialization. Journal of Cellular Physiology, 2011, 226(1): 224-235.

[28]Strasser P, Gimona M, Moessler H, Herzog M, Small J V. Mammalian calponin: identification and expression of genetic variants. FEBS Letters, 1993, 330(1): 13-18.

[29]Shibukawa Y, Yamazaki N, Kumasawa K, Daimon E, Tajiri M, Okada Y, Ikawa M, Wada Y. Calponin 3 regulates actin cytoskeleton rearrangement in trophoblastic cell fusion. Molecular Biology of the Cell, 2010, 21(15): 3973-3984.

[30]王书美, 张连峰. 碱性调宁蛋白的研究进展. 中国比较医学杂志, 2010, 20(1): 57-60.

Wang S M, Zhang L F. Advances on basic calponin. Chinese Journal of Comparative Medicine, 2010, 20(1): 57-60. (in Chinese)

[31]Su N, Chen M M, Chen S Y, Li C, Xie Y L, Zhu Y, Zhang Y Z, Zhao L, He Q F, Du X L, Chen D, Chen L. Overexpression of h1calponin in osteoblast lineage cells leads to a decreased in bone mass by disrupting osteoblast function and promoting osteoclast formation. Journal of Bone and Mineral Research, doi:[10.1002/jbmr.1778]
[1] YaTing JIA,HuiHui HU,YaJun ZHAI,Bing ZHAO,Kun HE,YuShan PAN,GongZheng HU,Li YUAN. Molecular Mechanism of Regulation by H-NS on IncFⅡ Plasmid Transmission of Multi-drug Resistant Chicken Escherichia coli [J]. Scientia Agricultura Sinica, 2022, 55(18): 3675-3684.
[2] Jie HU,XinYi WANG,Fei WANG. Functional Characterization of BmCaspase-8-Like (BmCasp8L) as an Immune Negative Regulatory Molecule in Silkworm (Bombyx mori) [J]. Scientia Agricultura Sinica, 2018, 51(21): 4188-4196.
[3] MA Xiao-Juan, LI Ya-Ming, HU Cui-Mei, WANG Fei, XIA Qing-You. Functional Characterization of BmFAF as an Immune Negative Regulatory Molecule in Silkworm (Bombyx mori) [J]. Scientia Agricultura Sinica, 2014, 47(15): 3085-3093.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备09089781号-30
Copyright 2018 © Editorial office of Scientia Agricultura Sinica
Tel: 86-010-82106279    E-mail: zgnykx@mail.caas.net.cn
Supported by Beijing Magtech Co.Ltd