Drp1 mediates compression-induced programmed necrosis of rat nucleus pulposus cells by promoting mitochondrial translocation of p53 and nuclear translocation of AIF

Oxidative stress and subsequent nucleus pulposus (NP) cell apoptosis are important contributors to the development of intervertebral disc degeneration (IDD). Emerging evidences show that long noncoding RNAs (lncRNAs) play a role in the pathogenesis of IDD. In this study, we investigated the role of lncRNA ANPODRT (anti-NP cell oxidative damage-related transcript) in oxidative stress and apoptosis in human NP cells. We found that ANPODRT was downregulated in degenerative NP tissues and in NP cells treated with tert-butyl hydroperoxide (TBHP, the oxidative stress inducer). ANPODRT overexpression alleviated oxidative stress and apoptosis in NP cells exposed to TBHP, while ANPODRT knockdown exerted opposing effects. Mechanistically, ANPODRT facilitated nuclear factor E2-related factor 2 (Nrf2) accumulation and nuclear translocation and activated its target genes by disrupting the kelch-like ECH-associated protein 1- (Keap1-) Nrf2 association in NP cells. Nrf2 knockdown abolished the antioxidative stress and antiapoptotic effects of ANPODRT in NP cells treated with TBHP. Collectively, our findings suggest that ANPODRT protects NP cells from oxidative stress and apoptosis, at least partially, by activating Nrf2 signaling, implying that ANPODRT may be a potential therapeutic target for IDD.

Download Full-text

Pterostilbene inhibits inflammation by promoting the nuclear translocation of Nrf2 in the rat nucleus pulposus

European Journal of Inflammation ◽

10.1177/1721727x18756536 ◽

2018 ◽

Vol 16 ◽

pp. 1721727X1875653

Author(s):

Soo-Min Lee ◽

Li-Yan Jiao ◽

Li-Bo Jiang ◽

Shu-Hao Liu ◽

Maka Lee ◽

...

Keyword(s):

Nitric Oxide ◽

Nucleus Pulposus ◽

Messenger Rna ◽

Nuclear Translocation ◽

Cell Counting ◽

Enzyme Linked Immunosorbent Assay ◽

Related Factor ◽

Nucleus Pulposus Cells ◽

Cox 2 ◽

Anti Inflammatory

Pterostilbene (PTE), a natural plant extract, has an anti-inflammatory effect; however, whether PTE could protect nucleus pulposus cells (NPCs) in the intervertebral disk from inflammation remains unclear. Primary NPCs isolated from Sprague-Dawley (SD) rats were cultured, and Cell Counting Kit-8 (CCK-8) analysis was used to test the cytotoxicity of PTE. The effect of PTE on interleukin-1β (IL-1β)-induced inflammation was analyzed using an enzyme-linked immunosorbent assay, real-time polymerase chain reaction (PCR), and a Griess test. Western blotting, immunofluorescence, and a nuclear factor erythroid 2-related factor 2 (Nrf2) small interfering RNA (siRNA) transfection were used to assess the involvement of Nrf2 in the anti-inflammatory mechanism of PTE on NPCs. The results of the CCK-8 analysis showed that PTE produced no cytotoxicity in NPCs at 20 μM for 24 h. PTE suppressed the production of nitric oxide (NO) and prostaglandin E2 (PGE2) and inhibited the messenger RNA (mRNA) expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) induced by IL-1β. PTE could promote the nuclear translocation of Nrf2 in NPCs. In addition, Nrf2 silence reversed the inhibitory effect of PTE on the production of NO and PGE2 and the expression of COX-2 and iNOS. These results indicate that PTE inhibits inflammation in the rat nucleus pulposus by promoting the nuclear translocation of Nrf2.

Download Full-text

Hydrogen peroxide induces programmed necrosis in rat nucleus pulposus cells through the RIP1/RIP3-PARP-AIF pathway

Journal of Orthopaedic Research® ◽

10.1002/jor.23751 ◽

2017 ◽

Cited By ~ 3

Author(s):

Lei Zhao ◽

Hui Lin ◽

Songfeng Chen ◽

Sheng Chen ◽

Min Cui ◽

...

Keyword(s):

Hydrogen Peroxide ◽

Nucleus Pulposus ◽

Nucleus Pulposus Cells ◽

Programmed Necrosis

Download Full-text

Reactive Oxygen Species Regulate Endoplasmic Reticulum Stress and ER-Mitochondrial Ca2+ Crosstalk to Promote Programmed Necrosis of Rat Nucleus Pulposus Cells under Compression

Oxidative Medicine and Cellular Longevity ◽

10.1155/2021/8810698 ◽

2021 ◽

Vol 2021 ◽

pp. 1-20

Author(s):

Hui Lin ◽

Yizhong Peng ◽

Jinye Li ◽

Zhe Wang ◽

Sheng Chen ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Endoplasmic Reticulum ◽

Nucleus Pulposus ◽

Molecular Mechanisms ◽

Therapeutic Targets ◽

Reactive Oxygen ◽

Oxygen Species ◽

Nucleus Pulposus Cells ◽

Programmed Necrosis ◽

Selective Channel

Programmed necrosis of nucleus pulposus (NP) cells caused by excessive compression is a crucial factor in the etiopathogenesis of intervertebral disc degeneration (IVDD). The endoplasmic reticulum (ER) and mitochondria are crucial regulators of the cell death signaling pathway, and their involvement in IVDD has been reported. However, the specific role of ER stress (ERS) and ER-mitochondria interaction in compression-induced programmed necrosis of NP cells remains unknown. Our studies revealed that compression enhanced ERS and the association between ER and mitochondria in NP cells. Suppression of ERS via 4-phenylbutyrate (4-PBA) or ER-mitochondrial Ca2+ crosstalk by inhibiting the inositol 1,4,5-trisphosphate receptor, glucose-regulated protein 75, voltage-dependent anion-selective channel 1 complex (IP3R–GRP75–VDAC1 complex) protected NP cells against programmed necrosis related to the poly(ADP-ribose) polymerase (PARP) apoptosis-inducing factor (AIF) pathway. Moreover, excessive reactive oxygen species are critical activators of ERS, leading to mitochondrial Ca2+ accumulation and consequent programmed necrosis. These data indicate that ERS and ER-mitochondrial Ca2+ crosstalk may be potential therapeutic targets for the treatment of IVDD-associated disorders. These findings provide new insights into the molecular mechanisms underlying IVDD and may provide novel therapeutic targets.

Download Full-text

Anisodamine Maintains the Stability of Intervertebral Disc Tissue by Inhibiting the Senescence of Nucleus Pulposus Cells and Degradation of Extracellular Matrix via Interleukin-6/Janus Kinases/Signal Transducer and Activator of Transcription 3 Pathway

Frontiers in Pharmacology ◽

10.3389/fphar.2020.519172 ◽

2020 ◽

Vol 11 ◽

Author(s):

Ning Tang ◽

Yulei Dong ◽

Chong Chen ◽

Hong Zhao

Keyword(s):

Intervertebral Disc ◽

Nucleus Pulposus ◽

Nuclear Translocation ◽

Telomerase Activity ◽

Nucleus Pulposus Cells ◽

Stat3 Pathway ◽

Stat3 Phosphorylation ◽

Ecm Degradation ◽

Collagen Ii ◽

Induced Apoptosis

Objectives: Anisodamine (ANI) has been used to treat a variety of diseases. However, the study of ANI in intervertebral disc degeneration (IVDD) is unclear. This study investigated the effects of ANI on degenerative nucleus pulposus cells (NPCs) and IVDD rats, and its possible mechanisms.Methods: Human nucleus pulposus cells (HNPCs) were treated with IL-1β (20 ng/ml) to simulate IVDD, and an IVDD rat model was constructed. IL-1β-induced HNPCs were treated with different concentrations (10, 20, or 40 μM) of ANI, and IVDD rats were also treated with ANI (1 mg/kg).Results: ANI treatment significantly reduced the apoptosis, caspase-3 and SA-β-gal activities, and p53 and p21 proteins expression, while promoted telomerase activity and aggrecan and collagen II synthesis in IL-1β-induced HNPCs. Moreover, the introduction of ANI inhibited the expression of IL-6, phosphorylation of JAK and STAT3, and nuclear translocation of p-STAT3 in Degenerated HNPCs. Additionally, the application of ANI abolished the effects of IL-6 on apoptosis, SA-β-gal and telomerase activity, and the expression of p53, p21, aggrecan and collagen II proteins in degenerated HNPCs. Simultaneously, ANI treatment enhanced the effects of AG490 (inhibitor of JAK/STAT3 pathway) on IL-1β-induced apoptosis, senescence and ECM degradation in HNPCs. Furthermore, ANI treatment markedly inhibited the apoptosis and senescence in the nucleus pulposus of IVDD rats, while promoted the synthesis of aggrecan and collagen II. ANI treatment obviously inhibited JAK and STAT3 phosphorylation and inhibited nuclear translocation of p-STAT3 in IVDD rats.Conclusion: ANI inhibited the senescence and ECM degradation of NPCs by regulating the IL-6/JAK/STAT3 pathway to improve the function of NPCs in IVDD, which may provide new ideas for the treatment of IVDD.

Download Full-text