The Effect of Hoxa11-as on the Differentiation of Osteoblasts in Osteoporosis by Regulating Cellular Autophagy

Yongcheng He; Peng Ling; Qunqiang Luo

doi:10.54097/ddkrdj46

Authors

Yongcheng He
Peng Ling
Qunqiang Luo

DOI:

https://doi.org/10.54097/ddkrdj46

Keywords:

Hoxa11-as, Osteoporosis, Autophagy, Osteoblasts

Abstract

Objective: To investigate the effect of long non-coding RNA (lncRNA) Hoxa11-as on the osteogenic differentiation in osteoporosis (OP) through the regulation of cellular autophagy. Methods: Establish a rat model of osteoporosis. Divide 12-week-old male rats into a blank group, a Hoxa11-as group, and a si-Hoxa11-as group, with 10 rats in each group. To establish glucocorticoid-induced osteoporosis, the required concentration is 1 ml/kg. The average body weight of this batch of rats is 500 g. The glucocorticoid is intraperitoneally injected at a dose of 0.5 ml/kg per day, and the intraperitoneal injection is continued for 8 weeks. After successfully establishing the rat model of osteoporosis, a bone rotary drill is used to create a 1-mm defect at the distal end of the femur of the model. At the same time, a hydrogel containing Hoxa11-as and si-Hoxa11-as plasmids is placed. After continuing the culture for 4 weeks, the samples are collected. Perform immunohistochemistry (IHC) and Real-time quantitative polymerase chain reaction (qPCR) experiments on the specimens to detect the protein expression levels of Hoxa11-as, LC3, Beclin1, and Runx2 factors. Results: The results of qPCR indicated that in the Hoxa11-as group, there were statistically significant differences in the mRNA expression levels of Hoxa11-as, LC3, Beclin1 and Runx2 (P < 0.05); in the si-Hoxa11-as group, there were also statistically significant differences in the mRNA expression levels of Hoxa11-as, LC3, Beclin1 and Runx2 (P < 0.05). Conclusion: Hoxa11-as has an inhibitory effect on cellular autophagy, while si-Hoxa11-as can activate cellular autophagy and enhance the differentiation level of osteoblasts.

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References

[1] Srivastava M, Deal C. Osteoporosis in elderly: prevention and treatment. Clin Geriatr Med. 2002 Aug;18(3):529-55.

[2] Xiao PL, Cui AY, Hsu CJ, et al. Global, regional prevalence, and risk factors of osteoporosis according to the World Health Organization diagnostic criteria: a systematic review and meta - analysis [J]. Osteoporos Int, 2022, 33 (10): 2137 - 2153.

[3] Yong EL, Logan S. Menopausal osteoporosis: screening, prevention and treatment. Singapore Med J. 2021 Apr;62 (4): 159-166.

[4] Li J, Chen X, Lu L, et al. The relationship between bone marrow adipose tissue and bone metabolism in postmenopausal osteoporosis [J]. Cytokine Growth Factor Rev, 2020, 52: 88 - 98.

[5] Deng P, Yuan Q, Cheng Y, et al. Loss of KDM4B exacerbates bone - fat imbalance and mesenchymal stromal cell exhaustion in skeletal aging [J]. Cell Stem Cell, 2021, 28 (6): 1057 - 1073.e7.

[6] Chen C, Huang L, Chen Y, et al. Hydrolyzed egg yolk peptide prevented osteoporosis by regulating Wnt/β - catenin signaling pathway in ovariectomized rats [J]. Sci Rep, 2024, 14 (1): 10227.

[7] Wang S, Deng Z, Ma Y, et al. The Role of Autophagy and Mitophagy in Bone Metabolic Disorders [J]. Int J Biol Sci, 2020, 16 (14): 2675 - 2691.

[8] Yoshida GJ. Therapeutic strategies of drug repositioning targeting autophagy to induce cancer cell death: from pathophysiology to treatment [J]. J Hematol Oncol, 2017, 10 (1): 67.

[9] Glick D, Barth S, Macleod KF. Autophagy: cellular and molecular mechanisms [J]. J Pathol, 2010, 221 (1): 3 - 12.

[10] Ding WX, Yin XM. Mitophagy: mechanisms, pathophysiological roles, and analysis [J]. Biol Chem, 2012, 393 (7): 547 - 564.

[11] Wang Y, Liu N, Lu B. Mechanisms and roles of mitophagy in neurodegenerative diseases [J]. CNS Neurosci Ther, 2019, 25 (7): 859 - 875.

[12] Wang Z, Liu N, Liu K, et al. Autophagy mediated CoCrMo particle - induced peri - implant osteolysis by promoting osteoblast apoptosis [J]. Autophagy, 2015, 11 (12): 2358 - 2369.

[13] Yang X, Yang J, Lei P, et al. LncRNA MALAT1 shuttled by bone marrow - derived mesenchymal stem cells - secreted exosomes alleviates osteoporosis through mediating microRNA - 34c/SATB2 axis [J]. Aging (Albany NY), 2019, 11 (20): 8777 - 8791.

[14] Zaidi M. Skeletal remodeling in health and disease [J]. Nat Med, 2007, 13 (7): 791 - 801.

[15] Kabeya Y, Mizushima N, Ueno T, et al. LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing [J]. EMBO J, 2000, 19 (21): 5720 - 5728.

[16] Pierrefite - Carle V, Santucci - Darmanin S, Breuil V, et al. Autophagy in bone: Self - eating to stay in balance [J]. Ageing Res Rev, 2015, 24 (Pt B): 206 - 217.

[17] Tanida I, Ueno T, Kominami E. LC3 conjugation system in mammalian autophagy [J]. Int J Biochem Cell Biol, 2004, 36 (12): 2503 - 2518.

[18] Li H, Li D, Ma Z, et al. Defective autophagy in osteoblasts induces endoplasmic reticulum stress and causes remarkable bone loss [J]. Autophagy, 2018, 14 (10): 1726 - 1741.

[19] Yin X, Zhou C, Li J, et al. Autophagy in bone homeostasis and the onset of osteoporosis [J]. Bone Res, 2019, 7: 28.

[20] Tran S, Fairlie WD, Lee EF. BECLIN1: Protein Structure, Function and Regulation [J]. Cells, 2021, 10 (6): 1522.

[21] Funderburk SF, Wang QJ, Yue Z. The Beclin 1 - VPS34 complex--at the crossroads of autophagy and beyond [J]. Trends Cell Biol, 2010, 20 (6): 355 - 362.

[22] Nishimura T, Tooze SA. Emerging roles of ATG proteins and membrane lipids in autophagosome formation [J]. Cell Discov, 2020, 6 (1): 32.

[23] Robinson JS, Klionsky DJ, Banta LM, et al. Protein sorting in Saccharomyces cerevisiae: isolation of mutants defective in the delivery and processing of multiple vacuolar hydrolases [J]. Mol Cell Biol, 1988, 8 (11): 4936 - 4948.

[24] Mei Y, Ramanathan A, Glover K, et al. Conformational Flexibility Enables the Function of a BECN1 Region Essential for Starvation - Mediated Autophagy [J]. Biochemistry, 2016, 55 (13): 1945 - 1958.

[25] Kim J, Kundu M, Viollet B, et al. AMPK and mTOR regulate autophagy through direct phosphorylation of Ulk1 [J]. Nat Cell Biol, 2011, 13 (2): 132 - 141.

[26] Ma X, Liu H, Murphy JT, et al. Regulation of the transcription factor EB - PGC1α axis by beclin - 1 controls mitochondrial quality and cardiomyocyte death under stress [J]. Mol Cell Biol, 2015, 35 (6): 956 - 976.

[27] Komori T. Whole Aspect of Runx2 Functions in Skeletal Development. Int J Mol Sci. 2022 May 21;23(10):5776.

[28] Lin TC. RUNX2 and Cancer. Int J Mol Sci. 2023 Apr 10;24 (8): 7001.

[29] Zhi F, Ding Y, Wang R, et al. Exosomal hsa_circ_0006859 is a potential biomarker for postmenopausal osteoporosis and enhances adipogenic versus osteogenic differentiation in human bone marrow mesenchymal stem cells by sponging miR - 431 - 5p [J]. Stem Cell Res Ther, 2021, 12 (1): 157.

[30] Cui Y, Fu S, Sun D, et al. EPC - derived exosomes promote osteoclastogenesis through LncRNA - MALAT1 [J]. J Cell Mol Med, 2019, 23 (6): 3843 - 3854.

[31] Cao G, Meng X, Han X, et al. Exosomes derived from circRNA Rtn4 - modified BMSCs attenuate TNF - α - induced cytotoxicity and apoptosis in murine MC3T3 - E1 cells by sponging miR - 146a [J]. Biosci Rep, 2020, 40 (5): BSR 20193436.

[32] Sun XJ, Wang Q, Guo B, et al. Identification of skin - related lncRNAs as potential biomarkers that involved in Wnt pathways in keloids [J]. Oncotarget, 2017, 8 (21): 34236 - 34244.

[33] Xue JY, Huang C, Wang W, et al. HOXA11 - AS: a novel regulator in human cancer proliferation and metastasis [J]. Onco Targets Ther, 2018, 11: 4387 - 4393.

[34] Lyu Y, Bai L, Qin C, et al. Long noncoding RNAs in neurodevelopment and Parkinson's disease [J]. Animal Model Exp Med, 2019, 2 (4): 239 - 251.