Experimental research on the effect of microRNA‑21 inhibitor on a rat model of intervertebral disc degeneration

Sheng,Xiaoming; Guo,Qingsong; Yu,Junbo; Xu,Youjia

doi:10.3892/etm.2018.6156

Experimental research on the effect of microRNA‑21 inhibitor on a rat model of intervertebral disc degeneration

Authors:
- Xiaoming Sheng
- Qingsong Guo
- Junbo Yu
- Youjia Xu
View Affiliations

Affiliations: Department of Orthopedics, The Affiliated Second Hospital of Soochow University, Suzhou, Jiangsu 215000, P.R. China, Department of General Surgery, The First Affiliated Hospital of Nantong University, Nantong, Jiangsu 225001, P.R. China, Department of Emergency, The First Affiliated Hospital of Nantong University, Nantong, Jiangsu 225001, P.R. China
Published online on: May 11, 2018 https://doi.org/10.3892/etm.2018.6156
Pages: 67-72
Copyright: © Sheng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Intervertebral disc degeneration is associated with angiogenesis and is the primary cause of disc‑associated disease. Several studies have indicated the importance of microRNA (miR)‑21 in angiogenesis. Thus, the present study aimed to validate the role and underlying mechanisms of miR‑21 in a rat model of intervertebral disc degeneration. A total of 60 specific‑pathogen‑free Sprague‑Dawley rats were used for in vivo experiments. A rat model of intervertebral disc degeneration was established and miR‑21 inhibitor (antagomiR‑21) was administered. The vertebral pulp and annulus fibrosus were isolated for immunohistochemical analysis of hypoxia inducible factor (HIF)‑1α and vascular endothelial growth factor (VEGF) expression. Lumbar spine proteoglycan content was detected with the phloroglucinol method. Disc cell apoptosis was detected with terminal deoxynucleotidyl‑transferase‑mediated dUTP nick end labeling staining. It was revealed that antagomiR‑21 treatment decreased the expression of HIF‑1α and VEGF in the vertebral pulp and annulus fibrosus. Furthermore, antagomiR‑21 treatment increased proteoglycan content and inhibited cell apoptosis in lumbar spines from model rats with intervertebral disc degeneration. In conclusion, antagomiR‑21 treatment exerted a protective role in a rat model of intervertebral disc degeneration, which may provide the basis for a potential therapeutic approach in the treatment of disc‑associated diseases.

View Figures

View References

1	Kang JD, Stefanovic-Racic M, Mcintyre LA, Georgescu HI and Evans CH: Toward a biochemical understanding of human intervertebral disc degeneration and herniation. Contributions of nitric oxide, interleukins, prostaglandin E2, and matrix metalloproteinases. Spine (Phila Pa 1976). 22:1065–1073. 1997. View Article : Google Scholar : PubMed/NCBI
2	Luoma K, Riihimäki H, Luukkonen R, Raininko R, Viikari-Juntura E and Lamminen A: Low back pain in relation to lumbar disc degeneration. Spine (Phila Pa 1976). 25:487–492. 2000. View Article : Google Scholar : PubMed/NCBI
3	Freemont AJ, Watkins A, Le Maitre C, Jeziorska M and Hoyland JA: Current understanding of cellular and molecular events in intervertebral disc degeneration: Implications for therapy. J Pathol. 196:374–379. 2002. View Article : Google Scholar : PubMed/NCBI
4	Fairbank J: Clinical importance of the intervertebral disc, or back pain for biochemists. Biochem Soc Trans. 30:829–831. 2002. View Article : Google Scholar : PubMed/NCBI
5	Adams MA and Roughley PJ: What is intervertebral disc degeneration, and what causes it? Spine (Phila Pa 1976). 31:2151–2161. 2006. View Article : Google Scholar : PubMed/NCBI
6	Binch ALA, Phillips KL, Chiverton N, Cole A, Michael AR, Breakwell L, Cross AK and Le Maitre CL: Role of semaphorins in angiogenesis and innervation in human intervertebral disc degeneration. Global Spine Journal. 04:1834–1841. 2014. View Article : Google Scholar
7	Ali R, Le-Maitre CL, Richardson SM, Hoyland JA and Freemont AJ: Connective tissue growth factor expression in human intervertebral disc: Implications for angiogenesis in intervertebral disc degeneration. Biotech Histochem. 83:239–245. 2008. View Article : Google Scholar : PubMed/NCBI
8	Lee JM, Song JY, Baek M, Jung HY, Kang H, Han IB, Kwon YD and Shin DE: Interleukin-1β induces angiogenesis and innervation in human intervertebral disc degeneration. J Orthop Res. 29:265–269. 2011. View Article : Google Scholar : PubMed/NCBI
9	Wang J, Chen H, Yuan W, Cao P, Shi L, Li R and Zang F: Analysis on angiogenesis in degenerative intervertebral disc and relevant factors. Zhonghua Guke Zazhi. 35:1200–1205. 2015.(In Chinese).
10	David G, Ciurea AV, Iencean SM and Mohan A: Angiogenesis in the degeneration of the lumbar intervertebral disc. J Med Life. 3:154–161. 2010.PubMed/NCBI
11	Calin GA and Croce CM: MicroRNA signatures in human cancers. Nat Rev Cancer. 6:857–866. 2006. View Article : Google Scholar : PubMed/NCBI
12	Zhao Y, Xu Y, Luo F, Xu W, Wang B, Pang Y, Zhou J, Wang X and Liu Q: Angiogenesis, mediated by miR-21, is involved arsenite-induced carcinogenesis. Toxicol Lett. 223:35–41. 2013. View Article : Google Scholar : PubMed/NCBI
13	Sabatel C, Malvaux L, Bovy N, Deroanne C, Lambert V, Gonzalez ML, Colige A, Rakic JM, Noël A, Martial JA and Struman I: MicroRNA-21 exhibits antiangiogenic function by targeting RhoB expression in endothelial cells. PLoS One. 6:e169792011. View Article : Google Scholar : PubMed/NCBI
14	Liu LZ, Li C, Chen Q, Jing Y, Carpenter R, Jiang Y, Kung HF, Lai L and Jiang BH: MiR-21 induced angiogenesis through AKT and ERK activation and HIF-1α expression. PLoS One. 6:e191392011. View Article : Google Scholar : PubMed/NCBI
15	Chen X, Liu J, He B, Li Y, Liu S, Wu B, Wang S, Zhang S, Xu X and Wang J: Vascular endothelial growth factor (VEGF) regulation by hypoxia inducible factor-1 alpha (HIF1A) starts and peaks during endometrial breakdown, not repair, in a mouse menstrual-like model. Hum Reprod. 30:2160–2170. 2015. View Article : Google Scholar : PubMed/NCBI
16	Wu WJ, Zhang XK, Zheng XF, Yang YH, Jiang SD and Jiang LS: SHH-dependent knockout of HIF-1 alpha accelerates the degenerative process in mouse intervertebral disc. Int J Immunopathol Pharmacol. 26:601–609. 2013. View Article : Google Scholar : PubMed/NCBI
17	Bayne K: Revised guide for the care and use of laboratory animals available. american physiological society. Physiologist. 39(199): 208–111. 1996.
18	Zhao CQ, Zhang YH, Jiang SD, Jiang LS and Dai LY: Both endoplasmic reticulum and mitochondria are involved in disc cell apoptosis and intervertebral disc degeneration in rats. Age (Dordr). 32:161–177. 2010. View Article : Google Scholar : PubMed/NCBI
19	Wu B, Meng C, Wang H, Jia C and Zhao Y: Proteoglycan and collagen type II in the adjacent intervertebral disc of the cervical instability models. Zhongguo Zuzhi Gongcheng Yanjiu. 17:5421–5426. 2013.(In Chinese).
20	Wei A, Brisby H, Chung SA and Diwan AD: Bone morphogenetic protein-7 protects human intervertebral disc cells in vitro from apoptosis. Spine J. 8:466–474. 2008. View Article : Google Scholar : PubMed/NCBI
21	Liu H, Huang X, Liu X, Xiao S, Zhang Y, Xiang T, Shen X, Wang G and Sheng B: miR-21 promotes human nucleus pulposus cell proliferation through PTEN/AKT signaling. Int J Mol Sci. 15:4007–4018. 2014. View Article : Google Scholar : PubMed/NCBI
22	Zhao D, Tu Y, Wan L, Bu L, Huang T, Sun X, Wang K and Shen B: In vivo monitoring of angiogenesis inhibition via down-regulation of mir-21 in a VEGFR2-luc murine breast cancer model using bioluminescent imaging. PLoS One. 8:e714722013. View Article : Google Scholar : PubMed/NCBI
23	Semenza GL: Vascular responses to hypoxia and ischemia. Arterioscler Thromb Vasc Biol. 30:648–652. 2010. View Article : Google Scholar : PubMed/NCBI
24	Wang GL, Jiang BH, Rue EA and Semenza GL: Hypoxia-inducible factor 1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O2 tension. Proc Natl Acad Sci USA. 92:5510–5514. 1995. View Article : Google Scholar : PubMed/NCBI
25	Rey S and Semenza GL: Hypoxia-inducible factor-1-dependent mechanisms of vascularization and vascular remodelling. Cardiovasc Res. 86:236–242. 2010. View Article : Google Scholar : PubMed/NCBI
26	Zhu L, Ye W and Cao Y: The relationship between the expression of cyclooxygenase-2, hypoxia-inducible factor-1α and angiogenesis in lumbar intervertebral disc degeneration of rats. Jiang Yaotong Zazhi. 33:174–178. 2012.(In Chinese).
27	Lyons G, Eisenstein SM and Sweet MB: Biochemical changes in intervertebral disc degeneration. Biochim Biophys Acta. 673:443–453. 1981. View Article : Google Scholar : PubMed/NCBI
28	Kohyama K, Saura R, Doita M and Mizuno K: Intervertebral disc cell apoptosis by nitric oxide: Biological understanding of intervertebral disc degeneration. Kobe J Med Sci. 46:283–295. 2000.PubMed/NCBI