scholarly journals LDHA-Mediated Glycolytic Metabolism in Nucleus Pulposus Cells Is a Potential Therapeutic Target for Intervertebral Disc Degeneration

2021 ◽  
Vol 2021 ◽  
pp. 1-14
Author(s):  
Longxi Wu ◽  
Jieliang Shen ◽  
Xiaojun Zhang ◽  
Zhenming Hu
Keyword(s):  
Energy Metabolism ◽  
Intervertebral Disc ◽  
Nucleus Pulposus ◽  
Disc Degeneration ◽  
Therapeutic Target ◽  
Intervertebral Disc Degeneration ◽  
Control Group ◽  
Potential Therapeutic Target ◽  
Strong Activity ◽  
Upstream Gene

The intervertebral disc degeneration (IDD) is considered to be an initiator of a series of spinal diseases, among which changes in the nucleus pulposus (NP) are the most significant. NP cells reside in a microenvironment with a lack of blood vessels, hypoxia, and low glucose within the intervertebral disc. Due to the strong activity of HIF-1α, glycolysis is the main pathway for energy metabolism in NP cells. Our previous study found that higher SIRT1 expression is beneficial to delay the degeneration of NP cells. In order to find the downstream genes by which SIRT1 acts on NP cells, we used iTRAQ sequencing to detect the differences between degenerated NP cells overexpressing SIRT1 and a control group (human NP cells were derived from surgery) and found that the expression of LDHA changed in the same direction with SIRT1. This suggests that SIRT1 may delay the degeneration of NP cells by regulating glycolysis. We then used a Seahorse XFe24 analyzer to measure the bioenergetic parameters of NP cells and obtained three findings: (a) glycolysis is the main energy metabolism pathway in NP cells, (b) there is a large difference in ATP production between senescent cells and young cells, and (c) SIRT1 can regulate the production of ATP from glycolysis by regulating LDHA. We also found that SIRT1 in NP cells has a positive regulatory effect on c-Myc which is an upstream gene of LDHA. Through observing IDD-related indicators such as apoptosis, proliferation, senescence, and extracellular matrix, we found that SIRT1 can delay degeneration, and interference with c-Myc and LDHA, respectively, weakens the protective effect of SIRT1. Interfering with LDHA alone can also inhibit glycolysis and accelerate degeneration. Overall, we found that the inhibition of glycolysis in Np cells significantly affects their normal physiological functions and determined that LDHA is a potential therapeutic target for the treatment of IDD.

scholarly journals Acidic pH promotes intervertebral disc degeneration: Acid-sensing ion channel -3 as a potential therapeutic target

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Hamish T. J. Gilbert ◽  
Nathan Hodson ◽  
Pauline Baird ◽  
Stephen M. Richardson ◽  
Judith A. Hoyland
Keyword(s):  
Intervertebral Disc ◽  
Ion Channel ◽  
Disc Degeneration ◽  
Therapeutic Target ◽  
Intervertebral Disc Degeneration ◽  
Acidic Ph ◽  
Potential Therapeutic Target

scholarly journals Knockout of miR-17-92 Cluster Protect Against Intervertebral Disc Degeneration by Inhibiting Apoptosis of Nucleus Pulposus Cells in Mice via the Activation of PI3K/Akt Pathway

2021 ◽  
Author(s):  
Yingjun Guo ◽  
Yang Meng ◽  
Hao Liu ◽  
Xiaofei Wang ◽  
Ying Hong ◽  
...  
Keyword(s):  
Intervertebral Disc ◽  
Nucleus Pulposus ◽  
Disc Degeneration ◽  
Intervertebral Disc Degeneration ◽  
Mrna Level ◽  
Akt Pathway ◽  
Functional Genes ◽  
Control Group ◽  
Expression Levels ◽  
Protein Levels

Abstract Background: microRNA(miR)-17-92 cluster is involved in a variety of physiological and pathological processes, and the purpose of this study is to preliminarily explore the role of miR-17-92 cluster in disc degeneration and the corresponding mechanisms.Methods: Hematoxylin and Eosin (HE) and Safranin O Staining were used to evaluate the degeneration of intervertebral disc. qRT-PCR was applied to evaluate the mRNA level of miR-17-92 cluster and functional genes of nucleus pulposus (NP) tissues, whose protein level was evaluated with Western-blot. Terminal-Deoxynucleoitidyl Transferase Mediated Nick End Labeling (TUNEL) was used to evaluate the apoptotic level of nucleus pulposus cell (NPC).Results: The expression levels of members of the miR-17-92 cluster were significantly increased in the NP tissues from patients with intervertebral disc degeneration (IVDD). Furthermore, in the 3-months and 24-months miR-17-92-ccKO mice, the degree of IVDD was significantly lower than that of the control group. At the same time, we also detected the expression levels of related functional genes in the NP tissues of mice in two groups. The results showed that the mRNA and protein levels of Bax and Caspase-3 in the knockout group were significantly lower than those in the control group, and the mRNA and protein levels of Bcl-2 were significantly higher. The TUNEL results showed that the apoptosis level of NPCs in the 3-month knockout mice was significantly lower than that in the control group. Finally, the assessment of pathway-related protein levels showed that p-Ser473-Akt expression ratio in the nucleus pulposus of mice in the knockout group were significantly increased, suggesting that the PI3K/Akt pathway was activated after miR-17-92 cluster knockout.Conclusion: To sum up, miR-17-92 cluster does play an important regulating role in IVDD, and the results showed that miR-17-92 cluster could inhibiting NPCs apoptosis by activating PI3K/Akt pathway, eventually producing protective effect against IVDD.

scholarly journals Expression of miR-195 and its target gene Bcl-2 in human intervertebral disc degeneration and their effects on nucleus pulposus cell apoptosis

2021 ◽  
Vol 16 (1) ◽  
Author(s):  
Xue-Lin Lin ◽  
Zhao-Yun Zheng ◽  
Qing-Shan Zhang ◽  
Zhen Zhang ◽  
You-Zhi An
Keyword(s):  
Cell Proliferation ◽  
Intervertebral Disc ◽  
Nucleus Pulposus ◽  
Disc Degeneration ◽  
Rat Model ◽  
Cell Apoptosis ◽  
Intervertebral Disc Degeneration ◽  
Target Gene ◽  
Blank Group ◽  
Tnf Α

Abstract Objective To investigate the expression of miR-195 and its target gene Bcl-2 in intervertebral disc degeneration (IVDD) and its effect on nucleus pulposus (NP) cell apoptosis. Methods The expressions of miR-195 and Bcl-2 in NP tissues of IVDD patients were quantified by qRT-PCR and western blotting, respectively. NP cells were divided into blank group, TNF-α group, TNF-α + miR-NC group, TNF-α + siBcl-2 group, and TNF-α + miR-195 inhibitors + siBcl-2 group. Cell proliferation was detected by MTT assay, cell apoptosis evaluated by flow cytometry, and mitochondrial membrane potential (MMP) tested by JC-1 staining. Moreover, the function of miR-195 on IVDD in vivo was investigated using a puncture-induced IVDD rat model. Results IVDD patients had significantly increased miR-195 expression and decreased Bcl-2 protein expression in NP tissues. The expression of miR-195 was negatively correlated with the expression of Bcl-2 in IVDD patients. Dual-luciferase reporter gene assay indicated that Bcl-2 was a target gene of miR-195. In comparison with blank group, TNF-α group showed decreased cell proliferation and MMP, increased cell apoptosis, upregulated expression of miR-195, Bax, and cleaved caspase 3, and downregulated Bcl-2 protein, while these changes were attenuated by miR-195 inhibitors. Additionally, siBcl-2 can reverse the protective effect of miR-195 inhibitors on TNF-α-induced NP cells. Besides, inhibition of miR-195 alleviated IVDD degeneration and NP cell apoptosis in the rat model. Conclusion MiR-195 was significantly upregulated in NP tissues of IVDD patients, and inhibition of miR-195 could protect human NP cells from TNF-α-induced apoptosis via upregulation of Bcl-2.

Sign in / Sign up

Export Citation Format

Share Document