CircGLCE alleviates intervertebral disc degeneration by regulating apoptosis and matrix degradation through the targeting of miR-587/STAP1

AbstractBackgroundIntervertebral disc degeneration (IDD) can induce profound global socioeconomic burdens. Recent studies have suggested that circular RNAs might have crucial functions in the progression of IDD. The purpose of this study was to identify a specific circular RNA and to investigate its regulatory mechanism in IDD.MethodsCircGLCE was selected after microarray analyses and was further analysed by RT-qPCR and FISH. After silencing CircGLCE, its function was assessed with RT-qPCR, immunofluorescence analysis and flow cytometry. Based on Sanger sequencing, miR-587 was identified as a direct target of CircGLCE, and it was further examined with RNA pulldown assays, RT-qPCR, dual luciferase assays and FISH. After silencing CircGLCE or miR-587, western blotting, immunofluorescence analysis, and flow cytometry were conducted. STAP1 was assessed by RT-qPCR and luciferase assay, and experiments with silenced and overexpressed miR-587 were performed. A rescue experiment was also included. In an IDD rat model, the in vivo effects of overexpressing CircGLCE on IDD were analysed with imaging techniques, TUNEL staining, FISH, western blotting, H&E staining and immunohistochemistry.ResultsCircGLCE was found to stably exist in the cytoplasm of nucleus pulposus (NP) cells. It was downregulated in IDD. Knockdown of CircGLCE promoted apoptosis and induced the expression of matrix-degrading enzymes in NP cells. CircGLCE served as a miR-587 sponge in NP cells. Inhibiting miR-587 counteracted the IDD-enhancing effect caused by silencing CircGLCE. STAP1 served as the miRNA target that mediated the functions of miR-587. Overexpressing CircGLCE alleviated IDD in vivo.ConclusionsCircGLCE attenuates IDD by regulating the apoptosis of NP cells and by regulating ECM degradation through the targeting of miR-587/STAP1. CircGLCE may be a potential therapeutic target for IDD treatments.

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CircERCC2 ameliorated intervertebral disc degeneration by regulating mitophagy and apoptosis through miR-182-5p/SIRT1 axis

Cell Death and Disease ◽

10.1038/s41419-019-1978-2 ◽

2019 ◽

Vol 10 (10) ◽

Cited By ~ 9

Author(s):

Lin Xie ◽

Weibo Huang ◽

Zhenhua Fang ◽

Fan Ding ◽

Fei Zou ◽

...

Keyword(s):

Intervertebral Disc ◽

Disc Degeneration ◽

Rat Model ◽

Intervertebral Disc Degeneration ◽

Circular Rnas ◽

Tert Butyl Hydroperoxide ◽

In Vitro Treatment

Abstract The molecular mechanism of intervertebral disc degeneration (IVDD) remains unclear. This study aimed to investigate the role of circular RNAs (circRNAs) in the pathogenesis of IVDD. We sued nucleus pulposus (NP) tissues of patients, tert-butyl hydroperoxide (TBHP) stimulated NP cells (NPCs), and IVDD rat model to explore the interaction between circERCC2 and miR-182-5p/SIRT1 axis. The results showed that downregulation of circERCC2 increased the level of miR-182-5p and decreased the level of SIRT1 in degenerative NP tissues in vivo as well as in TBHP-stimulated NPCs in vitro. Treatment of SIRT1-si activated apoptosis and inhibited mitophagy. Moreover, miR-182-5p-si could regulate the mitophagy and the apoptosis of NPCs by targeting SIRT1. The effects of circERCC2 on NPCs and IVDD rat model were mediated by miR-182-5p/SIRT1 axis. In conclusion, this study provides the first evidence that circERCC2 could ameliorate IVDD through miR-182-5p/SIRT1 axis by activating mitophagy and inhibiting apoptosis, and suggests that circERCC2 is a potentially effective therapeutic target for IVDD.

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An in vivo study of the effect of c-Jun on intervertebral disc degeneration in rats

Bioengineered ◽

10.1080/21655979.2021.1946459 ◽

2021 ◽

Vol 12 (1) ◽

pp. 4320-4330

Author(s):

Ming Lei ◽

Kangcheng Zhao ◽

Wenbin Hua ◽

Kun Wang ◽

Shuai Li ◽

...

Keyword(s):

Intervertebral Disc ◽

Disc Degeneration ◽

Intervertebral Disc Degeneration ◽

In Vivo Study

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Krüppel like factor 10 prevents intervertebral disc degeneration via TGF-β signaling pathway both in vitro and in vivo

Journal of Orthopaedic Translation ◽

10.1016/j.jot.2021.04.003 ◽

2021 ◽

Vol 29 ◽

pp. 19-29

Author(s):

Tongde Wu ◽

Xinhua Li ◽

Xuebing Jia ◽

Ziqi Zhu ◽

Jiawei Lu ◽

...

Keyword(s):

Intervertebral Disc ◽

Signaling Pathway ◽

Disc Degeneration ◽

Intervertebral Disc Degeneration

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Recovery of Intervertebral Disc Degeneration Post Enzymatic Treatment is Mediated by Matrix Metalloproteinases

ASME 2011 Summer Bioengineering Conference, Parts A and B ◽

10.1115/sbc2011-53887 ◽

2011 ◽

Author(s):

Nadeen Chahine ◽

Nate Stetson ◽

Neena Rajan ◽

Daniel Grande ◽

Mitchell Levine

Keyword(s):

Intervertebral Disc ◽

Disc Degeneration ◽

Enzymatic Degradation ◽

Intervertebral Disc Degeneration ◽

Biomechanical Properties ◽

Enzymatic Treatment ◽

Disc Injury ◽

Herniated Discs ◽

Dose Dependent

Enzymatic degradation of the intervertebral disc (IVD) with chondroitinase ABC (ChABC) reduces proteoglycan content of the IVD, thus simulating the GAG loss seen clinically in patients suffering from disc degeneration. This approach has been employed in models of disc injury in rats, rabbits and goats when administered over a large range of dosages [1–3]. Moreover, ChABC has also been used to induce repair of herniated discs in rabbits via chemonucleolysis [4, 5]. Despite the effectiveness of ChABC treatment to reduce the GAG content of the IVD, several recent studies including our own, have shown that this GAG loss is reversible at extended time points post enzymatic treatment [2,6,7]. The goal of the current study is to examine the dose dependent response of IVDs to degradation by ChABC in vivo. We hypothesize that administration of ChABC will result in dose dependent GAG loss and reduced mechanical properties. We administered ChABC at 0.1 U/ml, 1.0 U/ml and 10 U/ml and examined the changes in biomechanical properties, biochemical content, and gene expression in order to examine the biophysical and molecular mechanism by which GAG loss occurs in this model.

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Profiling and bioinformatics analysis of differentially expressed circular RNAs in human intervertebral disc degeneration

Acta Biochimica et Biophysica Sinica ◽

10.1093/abbs/gmz036 ◽

2019 ◽

Vol 51 (6) ◽

pp. 571-579 ◽

Cited By ~ 7

Author(s):

Shunmin Wang ◽

Jingchuan Sun ◽

Haisong Yang ◽

Weiguo Zou ◽

Bing Zheng ◽

...

Keyword(s):

Intervertebral Disc ◽

Disc Degeneration ◽

Intervertebral Disc Degeneration ◽

Expression Profiles ◽

Differentially Expressed ◽

Circular Rnas ◽

Functional Changes ◽

Qrt Pcr ◽

Reverse Transcription Pcr ◽

Microarray Expression

AbstractThe functional changes of nucleus pulposus (NP) cells are considered to be the initiating factors of intervertebral disc degeneration (IDD), and the differentially expressed circRNAs in NP cells may play an important role in the process of IDD. To identify circular RNAs (circRNAs) associated with human IDD, we isolated the NP cells from human degenerated and non-degenerated intervertebral disc and identified NP cells by microscopy and cell proliferation. CircRNA microarray expression profiles were obtained from NP cells of degenerated and non-degenerated intervertebral disc and further validated by quantitative reverse transcription PCR (qRT-PCR). The expression data were analyzed by bioinformatics. Microarray analysis identified 7294 circRNAs differentially expressed in degenerated human IDD NP cells. Among them, 3724 circRNAs were up-regulated and 3570 circRNAs were down-regulated by more than 2 folds. After validating by qRT-PCR, we predicted the possible miRNAs of the top dysregulated circRNAs using TargetScan, and miRanda. Furthermore, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that the most modulated circRNAs regulate the viability, degradation, apoptosis and oxidative stress in NP cells, and the possible mechanism underlying IDD was discussed. These results revealed that circRNAs may play a role in IDD and might be a promising candidate molecular target for gene therapy.

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A novel in vivo mouse intervertebral disc degeneration model induced by compressive suture

Experimental Cell Research ◽

10.1016/j.yexcr.2020.112359 ◽

2021 ◽

Vol 398 (1) ◽

pp. 112359

Author(s):

Zhuochao Liu ◽

Qi Zhou ◽

Jiancheng Zheng ◽

Changwei Li ◽

Weibin Zhang ◽

...

Keyword(s):

Intervertebral Disc ◽

Disc Degeneration ◽

Intervertebral Disc Degeneration

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Quercetin Suppresses Apoptosis and Attenuates Intervertebral Disc Degeneration via the SIRT1-Autophagy Pathway

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2020.613006 ◽

2020 ◽

Vol 8 ◽

Author(s):

Dong Wang ◽

Xin He ◽

Di Wang ◽

Pandi Peng ◽

Xiaolong Xu ◽

...

Keyword(s):

Intervertebral Disc ◽

Protective Effect ◽

Disc Degeneration ◽

Intervertebral Disc Degeneration ◽

Specific Effect ◽

Therapeutic Effects ◽

Dependent Manner ◽

Autophagy Pathway ◽

Dose Dependent

Intervertebral disc degeneration (IDD) has been generally accepted as the major cause of low back pain (LBP), which causes an enormous socioeconomic burden. Previous studies demonstrated that the apoptosis of nucleus pulposus (NP) cells and the dyshomeostasis of extracellular matrix (ECM) contributed to the pathogenesis of IDD, and effective therapies were still lacking. Quercetin, a natural flavonoid possessing a specific effect of autophagy stimulation and SIRT1 activation, showed some protective effect on a series of degenerative diseases. Based on previous studies, we hypothesized that quercetin might have therapeutic effects on IDD by inhibiting the apoptosis of NP cells and dyshomeostasis of ECM via the SIRT1-autophagy pathway. In this study, we revealed that quercetin treatment inhibited the apoptosis of NP cells and ECM degeneration induced by oxidative stress. We also found that quercetin promoted the expression of SIRT1 and autophagy in NP cells in a dose-dependent manner. Autophagy inhibitor 3-methyladenine (3-MA) reversed the protective effect of quercetin on apoptosis and ECM degeneration. Moreover, SIRT1 enzymatic activity inhibitor EX-527, suppressed quercetin-induced autophagy and the protective effect on NP cells, indicating that quercetin protected NP cells against apoptosis and prevented ECM degeneration via SIRT1-autophagy pathway. In vivo, quercetin was also demonstrated to alleviate the progression of IDD in rats. Taken together, our results suggest that quercetin prevents IDD by promoting SIRT1-dependent autophagy, indicating one novel and effective therapeutic method for IDD.

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