Effects of icariin on microRNAs expression in bone microvascular endothelial cells in steroids-induced femoral head lesions

doi:10.3969/j.issn.2095-4344.2016.15.002

Abstract

Abstract:

BACKGROUND: MicroRNAs (miRNAs) are widely involved in regulation of physiological processes, such as human development, cell proliferation, differentiation, and apoptosis, angiogenesis and lipid metabolism. MiRNAs also play an important regulating role in the pathological process of femoral head necrosis. At present, the research about the effect of icariin on miRNA expression in glucocorticoid- induced avascular necrosis is still in the exploratory stage, and the specific targets, possible regulation mechanism and signaling pathway remain unclear.
OBJECTIVE: To explore the effect of icariin on miRNA expression of bone microvascular endothelial cells in steroids-induced human femoral head lesions in vitro.
METHODS: Bone microvascular endothelial cells in cancellous bone of the femoral head were isolated and harvested in vitro. Icariin preconditioning preceded establishment of models of glucocorticoid-induced bone microvascular endothelial cell injury. Differential expression profiles and transcriptomes in glucocorticoid and normal groups were tested by miRNA microarrays. The most differentially expressed miR-23b and miR-339 in microarray analysis were further confirmed by real-time quantitative PCR, Meanwhile the effects of icariin on the expression of miR-23b-5p and miR-339-5p were detected.
RESULTS AND CONCLUSION: According to the microarray analysis, one miRNA was up-regulated and four mi RNAs were down-regulated in the glucocorticoid group (fold > 2, P < 0.05). Results of RT-qPCR revealed that miR-23b was down-regulated and miR-339 up-regulated in the glucocorticoid group, which were in agreement with the microarray analysis (P < 0.05). Icariin pretreatment effectively prevented the imbalances of miR-23b expression induced by glucocorticoid (P < 0.01). These findings indicate that Icariin may participate in the pathological process of steroid-induced femoral head necrosis through regulating the expression of miR-23b.
中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Microvessels, Endothelial Cells, Microchip Analytical Procedures, Tissue Engineering

Zhao Ding-yan, Guo Wan-shou, Yu Qing-sheng, Cheng Li-ming, Wang Bai-liang. Effects of icariin on microRNAs expression in bone microvascular endothelial cells in steroids-induced femoral head lesions[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(15): 2140-2147.

Figures/Tables 6

References

[1] Ikeuchi K, Hasegawa Y, Seki T, et al. Epidemiology of nontraumatic osteonecrosis of the femoral head in Japan. Mod Rheumatol. 2015;25(2):278-281.

[2] Mont MA, Cherian JJ, Sierra RJ, et al. Nontraumatic Osteonecrosis of the Femoral Head: Where Do We Stand Today? A Ten-Year Update. J Bone Joint Surg Am. 2015;97(19):1604-1627.

[3] Yuan HF, Von Roemeling C, Gao HD, et al. Analysis of altered microRNA expression profile in the reparative interface of the femoral head with osteonecrosis. Exp Mol Pathol. 2015;98(2):158-163.

[4] Nicoli S, Standley C, Walker P, et al. MicroRNA- mediated integration of haemodynamics and Vegf signalling during angiogenesis. Nature. 2010;464 (7292):1196-1200.

[5] Bartel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 2004;116(2): 281-297.

[6] Ambros V. The functions of animal microRNAs. Nature. 2004;431(7006):350-355.

[7] Inui M, Martello G, Piccolo S. MicroRNA control of signal transduction. Nat Rev Mol Cell Biol. 2010;11(4): 252-263.

[8] Gámez B, Rodriguez-Carballo E, Ventura F. MicroRNAs and post-transcriptional regulation of skeletal development. J Mol Endocrinol. 2014;52(3): R179-197.

[9] Mizuno Y, Yagi K, Tokuzawa Y, et al. miR-125b inhibits osteoblastic differentiation by down-regulation of cell proliferation. Biochem Biophys Res Commun. 2008; 368(2):267-272.

[10] [Rippe C, Blimline M, Magerko KA, et al. MicroRNA changes in human arterial endothelial cells with senescence: relation to apoptosis, eNOS and inflammation. Exp Gerontol. 2012;47(1):45-51.

[11] Kane NM, Thrasher AJ, Angelini GD, et al. Concise review: MicroRNAs as modulators of stem cells and angiogenesis. Stem Cells. 2014;32(5):1059-1066.

[12] 赵丁岩,俞庆声,郭万首,等.淫羊藿苷对激素诱导损伤人股骨头骨微血管内皮细胞蛋白质表达谱的影响[J].中华医学杂志,2016,96(13):1026-1030.

[13] Montgomery RL, van Rooij E. Therapeutic advances in MicroRNA targeting. J Cardiovasc Pharmacol. 2011; 57(1): 1-7.

[14] Nagpal JK, Rani R, Trink B, et al. Targeting miRNAs for drug discovery: a new paradigm. Curr Mol Med. 2010; 10(5):503-510.

[15] 路玉峰,俞庆声,郭万首,等.人股骨头骨微血管内皮细胞的分离培养方法[J].中国骨伤,2014,27(10):843-847.

[16] Hashemipour M, Dehkordi EH, Javanmard SH, et al. Von Willebrand factor, and soluble intercellular and vascular cell adhesion molecules as indices of endothelial activation in patients with congenital hypothyroidism. Horm Res Paediatr. 2011;76(2): 99-103.

[17] Gregg AJ, Schenkel AR. Cloning and structural analysis of equine platelet endothelial cell adhesion molecule (PECAM, CD31) and vascular cell adhesion molecule-1 (VCAM-1, CD106). Vet Immunol Immunopathol. 2008;122(3-4):295-308.

[18] Wang X, Qian W, Wu Z, et al. Preliminary screening of differentially expressed circulating microRNAs in patients with steroid?induced osteonecrosis of the femoral head. Mol Med Rep. 2014;10(6):3118-3124.

[19] Suzuki O, Bishop AT, Sunagawa T, et al. VEGF-promoted surgical angiogenesis in necrotic bone. Microsurgery. 2004;24(1):85-91.

[20] Yamasaki K, Nakasa T, Miyaki S, et al. Angiogenic microRNA-210 is present in cells surrounding osteonecrosis. J Orthop Res. 2012;30(8):1263-1270.

[21] Bouletreau PJ, Warren SM, Spector JA, et al. Hypoxia and VEGF up-regulate BMP-2 mRNA and protein expression in microvascular endothelial cells: implications for fracture healing. Plast Reconstr Surg. 2002;109(7):2384-2397.

[22] Suárez Y, Fernández-Hernando C, Pober JS, et al. Dicer dependent microRNAs regulate gene expression and functions in human endothelial cells. Circ Res. 2007;100(8):1164-1173.

[23] Kuehbacher A, Urbich C, Zeiher AM, et al. Role of Dicer and Drosha for endothelial microRNA expression and angiogenesis. Circ Res. 2007;101(1):59-68.

[24] Wang KC, Garmire LX, Young A, et al. Role of microRNA-23b in flow-regulation of Rb phosphorylation and endothelial cell growth Proc Natl Acad Sci U S A. 2010;107(7):3234-3239.

[25] Zhang H, Hao Y, Yang J, et al. Genome-wide functional screening of miR-23b as a pleiotropic modulator suppressing cancer metastasis. Nat Commun. 2011;2: 554.

[26] Majid S, Dar AA, Saini S, et al. miR-23b represses proto-oncogene Src kinase and functions as methylation-silenced tumor suppressor with diagnostic and prognostic significance in prostate cancer. Cancer Res. 2012;72(24):6435-6446.

[27] Zhou Q, Gallagher R, Ufret-Vincenty R, et al. Regulation of angiogenesis and choroidal neovascularization by members of microRNA- 23~27~24 clusters. Proc Natl Acad Sci USA. 2011; 108(20):8287-8292.

[28] Ham O, Song BW, Lee SY, et al. The role of microRNA-23b in the differentiation of MSC into chondrocyte by targeting protein kinase A signaling. Biomaterials. 2012;33(18):4500-4507.

[29] Ma Y, Yu S, Zhao W, et al. miR-27a regulates the growth, colony formation and migration of pancreatic cancer cells by targeting Sprouty2. Cancer Lett. 2010; 298(2):150-158.

[30] Walker JC, Harland RM. microRNA-24a is required to repress apoptosis in the developing neural retina. Genes Dev. 2009;23(9):1046-1051.

[31] Griffiths-Jones S, Saini HK, van Dongen S, et al. miRBase: tools for microRNA genomics. Nucleic Acids Res. 2008;36(Database issue):D154-158.