26RFa effects on osteogenic differentiation of human bone marrow mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.2014.10.006

Abstract

Abstract:

BACKGROUND: Studies have shown that 26RFa plays an important regulatory role in bone formation, pain, endocrine, cardiovascular disease and energy metabolism.
OBJECTIVE: To observe the effects of 26RFa on the proliferation and differentiation of human bone marrow mesenchymal stem cells.
METHODS: In order to obtain the most efficient concentration of 26RFa, human bone marrow mesenchymal stem cells were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide analysis. Cells were inoculated into 6-well plates and then divided into two groups: experimental group treated with
10-11 mol/L 26RFa and control group with no 26RFa. After 8, 12 and 16 days of osteogenic induction, alkaline phosphatase activities in induced cells were detected using alkaline phosphatase kit. After 21 and 28 days of osteogenic induction, alizarin red staining and Von Kossa staining were performed. The number of calcified nodules over each coverslip was calculated, and the expression of cbfa1 protein was detected by western blot assay.
RESULTS AND CONCLUSION: After 8, 12, and 16 days of osteogenic induction, the alkaline phosphatase activities were higher in the experimental group than the control group (P < 0.05, P < 0.01, P < 0.05). After 21 and 28 days of osteogenic induction, alizarin red staining and Von Kossa staining showed that the number of calcified nodules was higher in the experimental group than the control group, and the expression of cbfa1 protein was also higher in the experimental group (P < 0.05). These findings indicate that 26RFa can promote the osteogenic differentiation of human bone marrow mesenchymal stem cells under appropriate culture conditions.

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, alkaline phosphatase

CLC Number:

R394.2

Du Bin, Lin Qing, Liu Meng-jun, Chen Zhi-xin. 26RFa effects on osteogenic differentiation of human bone marrow mesenchymal stem cells [J]. Chinese Journal of Tissue Engineering Research, 2014, 18(10): 1508-1513.

Figures/Tables 3

References

[1]Erices A,Conget P,Minguell JJ.Mesenchymal progenitor cell in human umbilical cord blood.Br J Haematol.2000;109(1): 235-242.

[2]Bassett CA, Pilla AA, Pawluk RJ.A non-operative salvage of surgically-resisitant pseudarthroses and non-unions by pulsing electromagnetic fields. A preliminary report Clin Orthop Relat Res. 1977;(124):128-143.

[3]Basset CA, pawlik RJ, Pilla AA.Augmentation of bone repair by inductively coupled electromagnetic fields. Science. 1974; 184(4136):575-577.

[4]Gulotta LV, Kovacevic D, Packer JD, et al. Bone Marrow-Derived Mesenchymal Stem Cells Transduced With Scleraxis Improve Rotator Cuff Healing in a Rat Model. Am J Sports Med. 2011 Feb 18.

[5]Javazon EH, Colter DC, Schwarz EJ, et al. Rat marrow stromal cells are more sensitive to plating density and expand more rapidly from single-cell-derived colonies than human marrow stromal cells. Stem Cells. 2001;19(3):219-225.

[6]Minguell JJ,Erices A,Congetp.Mesenchymal stem Cell.Exp Biol Med.2001;226(6):507-526.

[7]Trivedi HL,Vanilar AV,Thakker U,et al.Human Adipose Tissue-derived Mesenchymal Stem Cells Combined with Hematopoietic Stem Cell Transplation Synthesize Insulin. Transplat Proc.2008;40(4):1135-1139.

[8]Yagi K,Kojima M,Oyagi S,et al.Application of Mesenchmal Stem Cell to Liver Regenerative Medicine. Yakugaku Zasshi. 2008;128(1):3-9.

[9]Sato K,Ozaki K,OH I,et,al.Nitric Oxide Plays a Critical Role in Suppression of T-cell Proliferation by Mesenchymal Stem Cell. Blood. 2007;109(1):228-234.

[10]Blazra BR,Taylor PA.Regulatiory T cells.Biol Blood Marrow Transplant.2005;11(2Sup12):46-49.

[11]Tang J, Wang J, Guo L,et al.Mesenchymal stem cells modified with stromal cell-derived factor 1 alpha improve cardiac remodeling via paracrine activation of hepatocyte growth factor in a rat model of myocardial infarction.Mol Cells.2010;29(1):9-19.

[12]Jeong WK, Park SW, Im GI. Growth factors reduce the suppression of proliferation and osteogenic ifferentiation by titanium particles on MSCs.J Biomed Mater Res A. 2008; 86(4):1137-1144.

[13]Ebra S，Nadayama K. Mechanism for the action of bone morphogenetic proteins and regulation of their activity. Spine.2002;27(16 suppl 1): 10-15.

[14]Bhatia M,Shokui S.Inflammatory Mediators in Acute Pancreatitis.Pathol.J Pathol.2000;190(2):117-125.

[15]刘震,程若川,方向,等. PAAF与ASPAAF诱导的肠损伤比较的实验研究[J].中国普外基础与临床杂志,2008,15(8):555-559.

[16]Boer HH, Schot LP, Veenstra JA, et al. Immunocytochemical identification of neural elements in the central nervous systems of a snail, some insects, a fish, and a mammal with an antiserum to the molluscan cardio-excitatory tetrapeptide FMRF-amide. Cell Tissue Res.1980;213(1): 21-27.

[17]Bock J,Liebisch G,Schweimer J,et al.Exogenous Sphingo2 Myelinase Causes Impaired Intestinal Epithelial Barrier Function. World Gastroenterol.2007;13(39):5217-5225.

[18]Zhang XP,Chen L,Hu QF,et al.Effects of Large Dose of Dexamethasone on Inflammatory Mediators and Pancreatic Cell Apoptosis of Rats with Severe Acute Pancreatitis. World J Gastroenterol. 2007;13(41):5506-5511

[19]Tsutsui K, Bentley GE, Kriegsfeld LJ,et al. Vaudry Discovery and evolutionary history of gonadotrophin-inhibitory hormone and kisspeptin: new key neuropeptides controlling reproduction. J Neuroendocrinol. 2010;22(7):716-727.

[20]Chartrel N, Dujardin C, Leprince J,et al. Isolation, characterization, and distribution of a novel neuropeptide, Rana RFamide (R-RFa), in the brain of the European green frog Rana esculenta. J Comp Neurol. 2002;448(2):111-127.

[21]Takayasu S, Sakurai T, Teranishi H ,et al.A neuropeptide ligand of the G protein-coupled receptor GPR103 regulates feeding,behavior arousal,and blood pressure in mice. Proc Natl Acad Sci U S A. 2006;103(19):7438-7443.

[22]Linacre A, Kellett E, Saunders S, et al. Burke Cardioactive neuropeptide Phe-Met-Arg-Phe-NH2 (FMRFamide) and novel related peptides are encoded in multiple copies by a single gene in the snail Lymnaea stagnalis.J Neurosci. 1990;10(2): 412-419.

[23]Fukusumi S, Yoshida H, Fujii R, et al. A new peptidic ligand and its receptor regulating adremal fuction in rats. J Biol Chem. 2003;278(47):46387-46395.

[24]Tobari Y, Iijima N, Tsunekawa K, et al.Identification, localisation and functional implication of 26RFa orthologue peptide in the brain of zebra finch (Taeniopygia guttata). J Neuroendocrinol. 2011;23(9):791-803.

[25]Liu Y, Zhang Y, Li S, et al. Molecular cloning and functional characterization of the first non-mammalian 26RFa/QRFP orthologue in Goldfish, Carassius auratus. Mol Cell Endocrinol. 2009;303(1-2):82-90.

[26]Yang HY, Tang J, Iadarola M, et al. Are Phe-Met-Arg-Phe-NH2 immunoreactive peptides endacoids modulating opiate antinociception? Prog Clin Biol Res. 1985; 192:313-322.

[27]Fukusumi S, Habata Y, Yoshida H,et al. Characteristics and distribution of endogenous RFamide-related peptide-1. Biochim Biophys Acta. 2001;1540(3):221-232.

[28]Chartrel N, Alonzeau J, Alexandre D, et al. Llorens-Cortes The RFamide neuropeptide 26RFa and its role in the control of neuroendocrine functions Front. Front Neuroendocrinol. 2011;32(4):387-397.

[29]Baribault H, Danao J, Gupte J, et al.The G-protein-coupled receptor GPR103 regulates bone formation. Mol Cell Biol. 2006;26(2):709-717.

[30]Ohtaki T, Shintani Y, Honda S,et al. Metastasis suppressor gene KiSS-1 encodes peptide ligand of a G-protein-coupled receptor.Nature.2001;411(6837): 613-617.

[31]Bruzzone F, Lectez B, Alexandre D,et al. Distribution of 26RFa binding sites and GPR103 mRNA in the central nervous system of the rat. J Comp Neurol. 2007;503(4): 573-591.

[32]Zhang Q, Qiu P, Arreaza MG, et al. P518/Qrfp sequence polymorphisms in SAMP6 osteopenic mouse. Genomics. 2007; 90(5):629-35

[33]王琳琳,董为人,朱衍菲,等.脂肪源性干细胞向心肌细胞分化及其移植治疗心血管疾病[J].中国组织工程研究与临床康复,2011, 15(14):2648-2652.

[34]Nakamichi I, Toivola DM, Strnad P, et al. Keratin 8 overexpression promotes mouse Mallory body formation. J Cell Biol. 2005;171(6):931-937.

[35]Ukena K, Tachibana T, Iwakoshi-Ukena E,et al. Identification, localization, and function of a novel avian hypothalamic neuropeptide, 26RFa, and its cognate receptor, G protein-coupled receptor-103. Endocrinology. 2010;151(5): 2255-2264.

[36]Casado-Díaz A, Santiago-Mora R, Jiménez R,et al. Cryopreserved human bone marrow mononuclear cells as a source of mesenchymal stromal cells: application in osteoporosis research. Cytotherapy. 2008;10(5):460-468.

[37]Matsumoto T,Kuroda R,Mifune Y,et al.Circulating endothelial/skeletal progenitor cells for bone regeneration and healing. Bone.2008;43(3):434-439.

[38]Lippens E, Cornelissen M.Slow Cooling Cryopreservation of Cell-Microcarrier Constructs. Cells Tissues Organs. 2010. [Epub ahead of print]

[39]Zajac JM, Mollereau C.RFamide peptides.Introduction. Peptides. 2006; 27(5):941-942.

[40]Bechtold DA, Luckman SM. The role of RFamide peptides in feeding. J Endocrinol. 2007;192(1):3-15.

[41]Jiang Y, Luo L, Gustafson EL, et al.Identification and characterization of a novel RF-amide pepfide ligand for orphan G-protein-coupled receptor SP9155. J Biol Chem. 2003;278(30):27652-27657.

[42]Mulumba M, Jossart C, Granata R, et al. GPR103b functions in the peripheral regulation of adipogenesis. Mol Endocrinol. 2010 ;24(8):1615-1625.

[43]Chartrel N, Dujardin C, Anouar Y,et al. Identification of 26RFa, a hypothalamic neuropeptide of the RFamide peptide family with orexigenic activity. Proc Natl Acad Sci U S A.2003; 100(25): 15247-15252.

[44]Bruzzone F, Lectez B, Tollemer H,et al.Anatomical distribution and biochemical characterization of the novel RFamide peptide 26RFa in the human hypothalamus and spinal cord．J Neurochem. 2006;99(2):616-627.