Mechanism by which miR-34a-5p/PLCD3 axis regulates osteoarthritis progression

doi:10.12307/2024.653

Abstract

Abstract: BACKGROUND: Molecular mechanisms targeting the miRNA/mRNA axis to regulate osteoarthritis disease process have been studied. We identified the mRNA: phospholipase C delta 3 (PLCD3) and its target miRNA(miR-34a-5p) with clinical predictive value through previous bioinformatics studies, while experiments to verify their specific roles and mechanisms in regulating osteoarthritis are still lacking.
OBJECTIVE: To investigate the regulatory role and mechanism of miR-34a-5p/PLCD3 axis on osteoarthritis progression.
METHODS: The synovium of 15 patients with knee osteoarthritis was selected as the osteoarthritis group, and the synovium of 15 young patients with internal fixation of patellar fracture caused by trauma during the same period was selected as the control group. The expression of PLCD3 and miR-34a-5p in the synovium was detected by real-time PCR. Human fibroblast like synovial cells-osteoarthritis (HFLS-OA) cells were treated by cell transfection and divided into miR-34a-5p mimic group, pCDH-PLCD3 group, miR-34a-5p mimic+pCDH-PLCD3 group, miR-34a-5p inhibitor group, si-PLCD3 group, and miR-34a-5p inhibitor+si-PLCD3 group. The relationship between PLCD3 and miR-34a-5p expression was detected by real-time PCR. The effects of HFLS-OA cell viability and cell migration in each group were detected by CCK-8 assay and cell scratch test. Western blot assay was used to detect the expression level of apoptosis marker protein. The expression of inflammatory factors was detected by ELISA.
RESULTS AND CONCLUSION: (1) PLCD3 was a direct target of miR-34a-5p, and the expression levels of PLCD3 and miR-34a-5p were negatively correlated. (2) Upregulation of PLCD3 promoted proliferation of HFLS-OA cells and inhibited cell migration. The up-regulation of miR-34a-5p significantly inhibited the activity of HFLS-OA cells and enhanced cell migration. Overexpression of miR-34a-5p significantly increased the levels of Casp3 and Casp9 proteins in HFLS-OA cells, while overexpression of PLCD3 showed the opposite trend. (3) PLCD3 overexpression significantly increased the expression of interleukin 6 and tumor necrosis factor alpha in HFLS-OA cells, while miR-34a-5p mimics showed protective activity. (4) The miR-34a-5p/PLCD3 axis may affect the progression of osteoarthritis by regulating the inflammatory process or apoptosis of synovial cells.

Key words: osteoarthritis, synovial membrane, miR-34a-5p, phospholipase Cδ3, PLCD3, interleukin 6, tumor necrosis factor alpha

CLC Number:

Ying Pu, Xu Yue, Lu Tong, Xue Yi, Miao Yiming. Mechanism by which miR-34a-5p/PLCD3 axis regulates osteoarthritis progression[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(33): 5320-5325.

Figures/Tables 7

References

[1] ABRAMOFF B, CALDERA FE. Osteoarthritis: Pathology, Diagnosis, and Treatment Options. Med Clin North Am. 2020;104(2):293-311.
[2] Foster NE, Eriksson L, Deveza L, et al. Osteoarthritis year in review 2022: Epidemiology & therapy. Osteoarthritis Cartilage. 2023; 31(7):876-883.
[3] De Roover A, Escribano-Núñez A, Monteagudo S, et al. Fundamentals of Osteoarthritis: Inflammatory mediators in Osteoarthritis. Osteoarthritis Cartilage. 2023;31(10):1303-1311.
[4] Cao X, Cui Z, Ding Z, et al. An osteoarthritis subtype characterized by synovial lipid metabolism disorder and fibroblast-like synoviocyte dysfunction. J Orthop Translat. 2022;33:142-152.
[5] Wu X, Sun AR, Crawford R, et al. Inhibition of Leukotriene A4 Hydrolase Suppressed Cartilage Degradation and Synovial Inflammation in a Mouse Model of Experimental Osteoarthritis. Cartilage. 2023: 19476035231169940.
[6] Chen X, Gong W, Shao X, et al. METTL3-mediated mA modification of ATG7 regulates autophagy-GATA4 axis to promote cellular senescence and osteoarthritis progression. Ann Rheum Dis. 2022;81(1): 87-99.
[7] Mimpen JY, Baldwin MJ, Cribbs AP, et al. Interleukin-17A Causes Osteoarthritis-Like Transcriptional Changes in Human Osteoarthritis-Derived Chondrocytes and Synovial Fibroblasts In Vitro. Front Immunol. 2021;12:676173.
[8] Park H, Lee HR, Shin HJ, et al. p16INK4a-siRNA nanoparticles attenuate cartilage degeneration in osteoarthritis by inhibiting inflammation in fibroblast-like synoviocytes.Biomater Sci. 2022;10(12): 3223-3235.
[9] Maglaviceanu A, Wu B, Kapoor M. Fibroblast-like synoviocytes: Role in synovial fibrosis associated with osteoarthritis. Wound Repair Regen. 2021;29(4):642-649.
[10] Lin L, Wen J, Lin B, et al. Phospholipase C Delta 3 inhibits apoptosis and promotes proliferation, migration, and invasion of thyroid cancer cells via Hippo pathway. Acta Biochim Biophys Sin (Shanghai). 2021;53(4):481-491.
[11] Dou XL, Xia FD, Li XY. Circ_0003747 promotes thyroid cancer progression by sponging miR-338-3p to upregulate PLCD3 expression. Epigenetics. 2023;18(1):2210339.
[12] Nakamura Y, Kanemaru K, Kojima R, et al. Simultaneous loss of phospholipase Cδ1 and phospholipase Cδ3 causes cardiomyocyte apoptosis and cardiomyopathy. Cell Death Dis. 2014;5(5):e1215.
[13] Ali SA, Espin-Garcia O, Wong AK, et al. Circulating microRNAs differentiate fast-progressing from slow-progressing and non-progressing knee osteoarthritis in the Osteoarthritis Initiative cohort. Ther Adv Musculoskelet Dis. 2022;14:1759720X221082917.
[14] Balaskas P, Goljanek-Whysall K, Clegg PD, et al. MicroRNA Signatures in Cartilage Ageing and Osteoarthritis. Biomedicines. 2023;11(4):1189.
[15] Kim M, Rubab A, Chan WCW, et al. Osteoarthritis year in review 2022: Genetics, genomics and epigenetics. Osteoarthritis Cartilage. 2023;31(7):865-875.
[16] 应璞, 路通, 许岳, 等.基于生物信息学挖掘骨关节炎潜在的关键生物标志物[J].国际检验医学杂志,2023,44(4):406-411.
[17] Song AF, Kang L, Wang YF, et al. MiR-34a-5p inhibits fibroblast‑like synoviocytes proliferation via XBP1. Eur Rev Med Pharmacol Sci. 2020; 24(22):11675-11682.
[18] Tian F, Wang J, Zhang Z, et al. LncRNA SNHG7/miR-34a-5p/SYVN1 axis plays a vital role in proliferation, apoptosis and autophagy in osteoarthritis. Biol Res. 2020;53(1):9.
[19] Endisha H, Datta P, Sharma A, et al. MicroRNA-34a-5p Promotes Joint Destruction During Osteoarthritis. Arthritis Rheumatol. 2021; 73(3):426-439.
[20] 中华医学会骨科学分会关节外科学组.骨关节炎诊疗指南(2018年版)[J].中华骨科杂志,2018,38(12):11.
[21] 中华医学会骨科学分会关节外科学组, 中国医师协会骨科医师分会骨关节炎学组, 国家老年疾病临床医学研究中心(湘雅医院), 等.中国骨关节炎诊疗指南(2021年版)[J].中华骨科杂志,2021, 41(18):24.
[22] Knights AJ, Redding SJ, Maerz T. Inflammation in osteoarthritis: the latest progress and ongoing challenges. Curr Opin Rheumatol. 2023;35(2):128-134.
[23] Ali SA, Peffers MJ, Ormseth MJ, et al. The non-coding RNA interactome in joint health and disease. Nat Rev Rheumatol. 2021; 17(11):692-705.
[24] Veronesi F, Costa V, Bellavia D, et al. Epigenetic Modifications of MiRNAs in Osteoarthritis: A Systematic Review on Their Methylation Levels and Effects on Chondrocytes, Extracellular Matrix and Joint Inflammation. Cells. 2023;12(14):1821.
[25] Feng L, Yang Z, Li Y, et al. MicroRNA-378 contributes to osteoarthritis by regulating chondrocyte autophagy and bone marrow mesenchymal stem cell chondrogenesis. Mol Ther Nucleic Acids. 2022;28:328-341.
[26] Coutinho de Almeida R, Ramos YFM, et al. RNA sequencing data integration reveals an miRNA interactome of osteoarthritis cartilage. Ann Rheum Dis. 2019;78(2):270-277.
[27] Chen HW, Huang CH, Huang CF, et al. Distinct subsets of synovial fibroblasts control cartilage destruction in joint diseases. Clin Exp Rheumatol. 2023 Sep 6. doi: 10.55563/clinexprheumatol/txl9rm.
[28] Maglaviceanu A, Wu B, Kapoor M. Fibroblast-like synoviocytes: Role in synovial fibrosis associated with osteoarthritis. Wound Repair Regen. 2021;29(4):642-649.
[29] Schuster R, Rockel JS, Kapoor M, et al. The inflammatory speech of fibroblasts. Immunol Rev. 2021;302(1):126-146.
[30] CHWASTEK J, KĘDZIORA M, BORCZYK M, et al. Inflammation-Driven Secretion Potential Is Upregulated in Osteoarthritic Fibroblast-Like Synoviocytes. Int J Mol Sci. 2022;23(19):11817.