Expression levels of miR-199 and TNF-α in the human degenerated intervertebral disc nucleus pulposus tissue and their biological effects
WANG Wei
GUO Zhao
YANG Lixin
Abstract：Objective To investigate the expression levels of miR-199 and tumor necrosis factor-α(TNF-α)in human degenerated intervertebral disc nucleus pulposus tissue and their biological effects.Methods Five tissue samples of degenerated intervertebral discs nucleus pulposus and 5 tissue samples of normal intervertebral discs nucleus pulposus form patients were collected.Differentially expressed microRNA(miRNA)between the two groups was screened by high-throughput sequencing.Changes in TNF-α level between groups were detected by enzyme-linked immunosorbent assay(ELISA),and miR-199 level was detected by reverse transcription-quantitative polymerase chain reaction(RT-qPCR).In vitro nucleus pulposus cell lines were cultured and induced with TNF-α(20ng/mL)to establish an in vitro apoptosis model.Nucleus pulposus cells were transfected with miR-control and miR-199 mimics,followed by TNF-α induction,thus exploring the regulatory effect of miR-199.Results Compared with normal nucleus pulposus tissue,miR-199 was significantly decreased in degenerated nucleus pulposus tissue,and TNF-α was significantly upregulated(P＜0.05).With the prolongation of TNF-α induction,the expression of miR-199 and the cell proliferation ability of nucleus pulposus cells decreased significantly(P＜0.05),and Lactate dehydrogenase(LDH)increased significantly(P＜0.05).Compared with the normal group,nucleus pulposus cells induced with TNF-α showed significantly lower viability(P＜0.05),and the expression of apoptosis-related proteins(Bax,Bcl-2,caspase-3 and cleaved caspase-3)were significantly increased(P＜0.05).While compared with the TNF-α group,the TNF-α+miR-199 mimics group reversed the experimental changes mentioned above,and LDH increased.Conclusion Down-regulated miR-199 and up-regulated TNF-α are seen in the nucleus pulposus tissue of degenerative intervertebral disc.Apoptosis of nucleus pulposus cells can be induced by TNF-α in a time-dependent manner,and TNF-α-induced apoptosis of nucleus pulposus cells can be reversed by overexpression of miR-199,suggesting that miR-199 participates in the onset of intervertebral disc degeneration(IDD)and can be used as an early diagnostic and therapeutic biomarker and therapeutic target for IDD.
Keywords：intervertebral disc degenerationmicroRNA(miRNA)tumor necrosis factor-αlpha(TNF-α)apoptosis
Publication Date：2025-02-28
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 212-217 )