昆明医科大学学报2025，Vol.46Issue(5)：21-29,9.DOI:10.12259/j.issn.2095-610X.S20250503
抑制miR-153-3p通过调控Nrf2延缓椎间盘退变的调节机制
Study on the Regulatory Mechanism of Inhibiting miR-153-3p to Delay Intervertebral Disc Degeneration via Nrf2 Regulation
摘要
Abstract
Objective To investigate the correlation mechanism between miR-153-3p and Nrf2 expression in human nucleus pulposus cells and intervertebral disc degeneration(IDD).Methods The oxidative damage model of nucleus pulposus cells was duplicated induced by H2O2.MiR-153-3p inhibitor-NC,si-NRF2-NC,miR-153-3p inhibitor,and si-NRF2 were transfected into nucleus pulposus cells according to the grouping require.The transfection efficiency was detected by RT-qPCR and Western blot.The cell viability was determined by the CCK-8 assay,when the intracellular reactive oxygen species(ROS)levels and the ratio of mitochondrial membrane potential decline were measured by flow cytometry,RT-qPCR was used to detect the expression levels of Nrf2,MMP-3,Col II,PINK1,Parkin,P62,and p38 MAPK.And dual luciferase reporter assay was used to measure luciferase activity.Results(1)HNPCs treated with H2O2 showed a decrease in HNPCs cell viability,a reduction in mitochondrial membrane potential,an increase in ROS levels,and a decrease in Col II expression(P<0.05).And the expression of MMP-3,P62,and p38 MAPK increased,while the expression of PINK1 and Parkin decreased.There was no significant change in Nrf2(P>0.05).(2)Inhibition of miR-153-3p expression in nucleus pulposus cells treated with H2O2 led to increased cell viability,elevated mitochondrial membrane potential,reduced ROS levels,and enhanced Col-II expression,accompanied by decreased expression of MMP-3,P62,and p38 MAPK,while simultaneously increasing the expression of PINK1,Parkin,and Nrf2(P<0.05).(3)When miR-153-3p expression was inhibited and Nrf2 expression was silenced in nucleus pulposus cells treated with H2O2,a notable decline in cell viability and mitochondrial membrane potential was observed,along with a marked increase in ROS levels.Additionally,Col-II expression decreased,whereas the expression of MMP-3,P62,and p38 MAPK increased.However,the expression of Nrf2,PINK1,and Parkin decreased(P<0.05).(4)Dual-luciferase assay analysis revealed binding sites and a binding relationship between miR-153-3p and Nrf2,indicating a negative correlation between miR-153-3p and Nrf2(P<0.05).Conclusion Inhibition of miR-153-3p expression can alleviate H2O2 induced degeneration and fibrosis of nucleus pulposus cells,activate autophagy of damaged nucleus pulposus cells,and delay apoptosis of nucleus pulposus cells.This effect is related to the PINK1/Parkin pathway and p38 MAPK inflammatory response pathway regulated by Nrf2.关键词
椎间盘退变/人髓核细胞/miR-153-3p/Nrf2Key words
Intervertebral disc degeneration/Human nucleus pulposus cells/miR-153-3p/Nrf2分类
临床医学引用本文复制引用
庾珊,肖林,龚东平,梁楼峰,许夏懿,梁华新,肖世海..抑制miR-153-3p通过调控Nrf2延缓椎间盘退变的调节机制[J].昆明医科大学学报,2025,46(5):21-29,9.基金项目
广西中医药管理局科研基金资助项目(GXZYZ20210506) （GXZYZ20210506）
广西壮族自治区中医药局2021年自筹经费科研课题(20210630) （20210630）
广西中医药重点学科建设项目(GZXK-Z-20-62) （GZXK-Z-20-62）
