Knockdown of exosome‑mediated lnc‑PVT1 alleviates lipopolysaccharide‑induced osteoarthritis progression by mediating the HMGB1/TLR4/NF‑κB pathway via miR‑93‑5p

Meng,Yong; Qiu,Siqiang; Sun,Long; Zuo,Jinliang

doi:10.3892/mmr.2020.11594

Knockdown of exosome‑mediated lnc‑PVT1 alleviates lipopolysaccharide‑induced osteoarthritis progression by mediating the HMGB1/TLR4/NF‑κB pathway via miR‑93‑5p

Authors:
- Yong Meng
- Siqiang Qiu
- Long Sun
- Jinliang Zuo
View Affiliations

Affiliations: Qingdao University, Qingdao, Shandong 266000, P.R. China, Department of Spine Surgery, The Fourth People's Hospital of Jinan, Jinan, Shandong 250031, P.R. China, Department of Orthopedics, Weihai Municipal Hospital, Weihai, Shandong 264200, P.R. China
Published online on: October 14, 2020 https://doi.org/10.3892/mmr.2020.11594
Pages: 5313-5325
Copyright: © Meng 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

Osteoarthritis is a chronic degenerative joint disease. Long non‑coding RNA plasmacytoma variant translocation 1 (PVT1) is involved in the progression of osteoarthritis and exosomes serve a central role in intercellular communication. However, whether PVT1 can be mediated by exosomes in osteoarthritis has not been reported. Whole blood was drawn from osteoarthritis patients and healthy volunteers. Lipopolysaccharide (LPS) was used to stimulate human normal chondrocytes (C28/I2) to construct a cell damage model in vitro. Protein levels were examined via western blot analysis. eThe expression of PVT1, microRNA (miR)‑93‑5p and high mobility groupprotein B1 (HMGB1) was evaluated through reverse transcription‑quantitative PCR. Cell viability and apoptosis were determined through CCK‑8 or flow cytometric assay. Inflammatory cytokines were measured via ELISA. The relationship between PVT1 or HMGB1 and miR‑93‑5p was confirmed by dual‑luciferase reporter assay. PVT1, HMGB1 and exosomal PVT1 were upregulated while miR‑93‑5p was downregulated in osteoarthritis patient serum and LPS‑induced C28/I2 cells. Exosomes from osteoarthritis patient serum and LPS‑treated C28/I2 cells increased PVT1 expression in C28/I2 cells. PVT1 depletion reversed the decrease of viability and the increase of apoptosis, inflammation responses and collagen degradation of C28/I2 cells induced by LPS. PVT1 regulated HMGB1 expression via sponging miR‑93‑5p. miR‑93‑5p inhibition abolished PVT1 silencing‑mediated viability, apoptosis, inflammation responses and collagen degradation of LPS‑stimulated C28/I2 cells. HMGB1 increase overturned miR‑93‑5p upregulation‑mediated viability, apoptosis, inflammation responses and collagen degradation of LPS‑stimulated C28/I2 cells. Furthermore, PVT1 modulated the Toll‑like receptor 4/NF‑κB pathway through an miR‑93‑5p/HMGB1 axis. In summary, exosome‑mediated PVT1 regulated LPS‑induced osteoarthritis progression by modulating the HMGB1/TLR4/NF‑κB pathway via miR‑93‑5p, providing a new route for possible osteoarthritis treatment.

View Figures

View References

1	Hussain SM, Neilly DW, Baliga S, Patil S and Meek R: Knee osteoarthritis: A review of management options. Scott Med J. 61:7–16. 2016.PubMed/NCBI
2	Kurtz SM, Ong KL, Lau E, Widmer M, Maravic M, Gómez-Barrena E, de Pina Mde F, Manno V, Torre M, Walter WL, et al: International survey of primary and revision total knee replacement. Int Orthop. 35:1783–1789. 2011.PubMed/NCBI
3	Zhen G, Wen C, Jia X, Li Y, Crane JL, Mears SC, Askin FB, Frassica FJ, Chang W, Yao J, et al: Inhibition of TGF-β signaling in mesenchymal stem cells of subchondral bone attenuates osteoarthritis. Nat Med. 19:704–712. 2013.PubMed/NCBI
4	Taylor N: Nonsurgical management of osteoarthritis knee pain in the older adult. Clin Geriatr Med. 33:41–51. 2017.PubMed/NCBI
5	Batista PJ and Chang HY: Long noncoding RNAs: Cellular address codes in development and disease. Cell. 152:1298–1307. 2013.PubMed/NCBI
6	Schmitz SU, Grote P and Herrmann BG: Mechanisms of long noncoding RNA function in development and disease. Cell Mol Life Sci. 73:2491–2509. 2016.PubMed/NCBI
7	Liu Q, Hu X, Zhang X, Dai L, Duan X, Zhou C and Ao Y: The TMSB4 pseudogene lncRNA functions as a competing endogenous RNA to promote cartilage degradation in human osteoarthritis. Mol Ther. 24:1726–1733. 2016.PubMed/NCBI
8	Zhang Y, Wang F, Chen G, He R and Yang L: LncRNA MALAT1 promotes osteoarthritis by modulating miR-150-5p/AKT3 axis. Cell Biosci. 9:542019.PubMed/NCBI
9	Cao L, Wang Y, Wang Q and Huang J: LncRNA FOXD2-AS1 regulates chondrocyte proliferation in osteoarthritis by acting as a sponge of miR-206 to modulate CCND1 expression. Biomed Pharmacother. 106:1220–1226. 2018.PubMed/NCBI
10	Cui M, You L, Ren X, Zhao W, Liao Q and Zhao Y: Long non-coding RNA PVT1 and cancer. Biochem Biophys Res Commun. 471:10–14. 2016.PubMed/NCBI
11	Huang W, Lan X, Li X, Wang D, Sun Y, Wang Q, Gao H and Yu K: Long non-coding RNA PVT1 promote LPS-induced septic acute kidney injury by regulating TNFα and JNK/NF-κB pathways in HK-2 cells. Int Immunopharmacol. 47:134–140. 2017.PubMed/NCBI
12	Feng F, Qi Y, Dong C and Yang C: PVT1 regulates inflammation and cardiac function via the MAPK/NF-κB pathway in a sepsis model. Exp Ther Med. 16:4471–4478. 2018.PubMed/NCBI
13	Li Y, Li S, Luo Y, Liu Y and Yu N: LncRNA PVT1 regulates chondrocyte apoptosis in osteoarthritis by acting as a sponge for miR-488-3p. DNA Cell Biol. 36:571–580. 2017.PubMed/NCBI
14	Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004.PubMed/NCBI
15	Krol J, Loedige I and Filipowicz W: The widespread regulation of microRNA biogenesis, function and decay. Nat Rev Genet. 11:597–610. 2010.PubMed/NCBI
16	Wang X, Liao Z, Bai Z, He Y, Duan J and Wei L: MiR-93-5p promotes cell proliferation through down-regulating PPARGC1A in hepatocellular carcinoma cells by bioinformatics analysis and experimental verification. Genes (Basel). 9:512018.
17	Xiang Y, Liao XH, Yu CX, Yao A, Qin H, Li JP, Hu P, Li H, Guo W, Gu CJ and Zhang TC: MiR-93-5p inhibits the EMT of breast cancer cells via targeting MKL-1 and STAT3. Exp Cell Res. 357:135–144. 2017.PubMed/NCBI
18	Zhang Y, Wei QS, Ding WB, Zhang LL, Wang HC, Zhu YJ, He W, Chai YN and Liu YW: Increased microRNA-93-5p inhibits osteogenic differentiation by targeting bone morphogenetic protein-2. PLoS One. 12:e01826782017.PubMed/NCBI
19	Hua Q, Chen Y, Liu Y, Li M, Diao Q, Xue H, Zeng H, Huang L and Jiang Y: Circular RNA 0039411 is involved in neodymium oxide-induced inflammation and antiproliferation in a human bronchial epithelial cell line via sponging miR-93-5p. Toxicol Sci. 170:69–81. 2019.PubMed/NCBI
20	Xue H, Tu Y, Ma T, Wen T, Yang T, Xue L, Cai M, Wang F and Guan M: miR-93-5p attenuates IL-1β-induced chondrocyte apoptosis and cartilage degradation in osteoarthritis partially by targeting TCF4. Bone. 123:129–136. 2019.PubMed/NCBI
21	Harding CV, Heuser JE and Stahl PD: Exosomes: Looking back three decades and into the future. J Cell Biol. 200:367–371. 2013.PubMed/NCBI
22	Valadi H, Ekström K, Bossios A, Sjöstrand M, Lee JJ and Lötvall JO: Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat Cell Biol. 9:654–659. 2007.PubMed/NCBI
23	Lo Cicero A, Stahl PD and Raposo G: Extracellular vesicles shuffling intercellular messages: For good or for bad. Curr Opin Cell Biol. 35:69–77. 2015.PubMed/NCBI
24	Buzas EI, György B, Nagy G, Falus A and Gay S: Emerging role of extracellular vesicles in inflammatory diseases. Nat Rev Rheumatol. 10:356–364. 2014.PubMed/NCBI
25	Li ZL, Lv LL, Tang TT, Wang B, Feng Y, Zhou LT, Cao JY, Tang RN, Wu M, Liu H, et al: HIF-1α inducing exosomal microRNA-23a expression mediates the cross-talk between tubular epithelial cells and macrophages in tubulointerstitial inflammation. Kidney Int. 95:388–404. 2019.PubMed/NCBI
26	Patel NA, Moss LD, Lee JY, Tajiri N, Acosta S, Hudson C, Parag S, Cooper DR, Borlongan CV and Bickford PC: Long noncoding RNA MALAT1 in exosomes drives regenerative function and modulates inflammation-linked networks following traumatic brain injury. J Neuroinflammation. 15:2042018.PubMed/NCBI
27	Zhang HG, Liu C, Su K, Yu S, Zhang L, Zhang S, Wang J, Cao X, Grizzle W and Kimberly RP: A membrane form of TNF-alpha presented by exosomes delays T cell activation-induced cell death. J Immunol. 176:7385–7393. 2006.PubMed/NCBI
28	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001.PubMed/NCBI
29	Cao C, Yin C, Shou S, Wang J, Yu L, Li X and Chai Y: Ulinastatin protects against LPS-induced acute lung injury by attenuating TLR4/NF-κB pathway activation and reducing inflammatory mediators. Shock. 50:595–605. 2018.PubMed/NCBI
30	Hare KB, Stefan Lohmander L, Kise NJ, Risberg MA and Roos EM: Middle-aged patients with an MRI-verified medial meniscal tear report symptoms commonly associated with knee osteoarthritis. Acta Orthop. 88:664–669. 2017.PubMed/NCBI
31	Huang ZY, Stabler T, Pei FX and Kraus VB: Both systemic and local lipopolysaccharide (LPS) burden are associated with knee OA severity and inflammation. Osteoarthritis Cartilage. 24:1769–1775. 2016.PubMed/NCBI
32	Zhao C, Wang Y, Jin H and Yu T: Knockdown of microRNA-203 alleviates LPS-induced injury by targeting MCL-1 in C28/I2 chondrocytes. Exp Cell Res. 359:171–178. 2017.PubMed/NCBI
33	Sun T, Yu J, Han L, Tian S, Xu B, Gong X, Zhao Q and Wang Y: Knockdown of long non-coding RNA RP11-445H22.4 alleviates LPS-induced injuries by regulation of miR-301a in osteoarthritis. Cell Physiol Biochem. 45:832–843. 2018.PubMed/NCBI
34	Li F, Sun J, Huang S, Su G and Pi G: LncRNA GAS5 overexpression reverses LPS-induced inflammatory injury and apoptosis through up-regulating KLF2 expression in ATDC5 chondrocytes. Cell Physiol Biochem. 45:1241–1251. 2018.PubMed/NCBI
35	Chen WK, Yu XH, Yang W, Wang C, He WS, Yan YG, Zhang J and Wang WJ: lncRNAs: Novel players in intervertebral disc degeneration and osteoarthritis. Cell Prolif. 50:e123132017.
36	Xu J and Xu Y: The lncRNA MEG3 downregulation leads to osteoarthritis progression via miR-16/SMAD7 axis. Cell Biosci. 7:692017.PubMed/NCBI
37	Barile L and Vassalli G: Exosomes: Therapy delivery tools and biomarkers of diseases. Pharmacol Ther. 174:63–78. 2017.PubMed/NCBI
38	Zhao Y, Zhao J, Guo X, She J and Liu Y: Long non-coding RNA PVT1, a molecular sponge for miR-149, contributes aberrant metabolic dysfunction and inflammation in IL-1β-simulated osteoarthritic chondrocytes. Biosci Rep. 38:BSR201805762018.PubMed/NCBI
39	Ding LB, Li Y, Liu GY, Li TH, Li F, Guan J and Wang HJ: Long non-coding RNA PVT1, a molecular sponge of miR-26b, is involved in the progression of hyperglycemia-induced collagen degradation in human chondrocytes by targeting CTGF/TGF-β signal ways. Innate Immun. 26:204–214. 2020.PubMed/NCBI
40	Liu J, Jiang M, Deng S, Lu J, Huang H, Zhang Y, Gong P, Shen X, Ruan H, Jin M and Wang H: miR-93-5p-containing exosomes treatment attenuates acute myocardial infarction-induced myocardial damage. Mol Ther Nucleic Acids. 11:103–115. 2018.PubMed/NCBI
41	Laursen TL, Støy S, Deleuran B, Vilstrup H, Grønbaek H and Sandahl TD: The damage-associated molecular pattern HMGB1 is elevated in human alcoholic hepatitis, but does not seem to be a primary driver of inflammation. APMIS. 124:741–747. 2016.PubMed/NCBI
42	Srikrishna G and Freeze HH: Endogenous damage-associated molecular pattern molecules at the crossroads of inflammation and cancer. Neoplasia. 11:615–628. 2009.PubMed/NCBI
43	Qin Y, Chen Y, Wang W, Wang Z, Tang G, Zhang P, He Z, Liu Y, Dai SM and Shen Q: HMGB1-LPS complex promotes transformation of osteoarthritis synovial fibroblasts to a rheumatoid arthritis synovial fibroblast-like phenotype. Cell Death Dis. 5:e10772014.PubMed/NCBI
44	Wang X, Guo Y, Wang C and Yu H, Yu X and Yu H: MicroRNA-142-3p inhibits chondrocyte apoptosis and inflammation in osteoarthritis by targeting HMGB1. Inflammation. 39:1718–1728. 2016.PubMed/NCBI
45	Liang S, Lv ZT, Zhang JM, Wang YT, Dong YH, Wang ZG, Chen K, Cheng P, Yang Q, Guo FJ, et al: Necrostatin-1 attenuates trauma-induced mouse osteoarthritis and IL-1β induced apoptosis via HMGB1/TLR4/SDF-1 in primary mouse chondrocytes. Front Pharmacol. 9:13782018.PubMed/NCBI
46	Jiang Y, Zhu L, Zhang T, Lu H, Wang C, Xue B, Xu X, Liu Y, Cai Z, Sang W, et al: BRD4 has dual effects on the HMGB1 and NF-κB signalling pathways and is a potential therapeutic target for osteoarthritis. Biochim Biophys Acta Mol Basis Dis. 1863:3001–3015. 2017.PubMed/NCBI