Study on the Regulatory Mechanism of Inhibiting miR-153-3p to Delay Intervertebral Disc Degeneration via Nrf2 Regulation

Shan YU; Lin XIAO; Dongping GONG; Loufeng LIANG; Xiayi XU; Huaxin LIANG; Shihai XIAO

Turn off MathJax

Article Contents

Abstract

References

Relative Articles

Article Navigation > Journal of Kunming Medical University > 2025 > Accepted Manuscript

Shan YU, Lin XIAO, Dongping GONG, Loufeng LIANG, Xiayi XU, Huaxin LIANG, Shihai XIAO. Study on the Regulatory Mechanism of Inhibiting miR-153-3p to Delay Intervertebral Disc Degeneration via Nrf2 Regulation[J]. Journal of Kunming Medical University.

Citation:

PDF( 1036 KB)

Study on the Regulatory Mechanism of Inhibiting miR-153-3p to Delay Intervertebral Disc Degeneration via Nrf2 Regulation

1.
Dept. of Pain Management，International Zhuang Medicine Hospital Affiliated to Guangxi University of Traditional Chinese Medicine，Nanning Guangxi 530201
2.
Guangxi University of Traditional Chinese Medicine ，Nanning Guangxi 530200，China

Received Date: 2024-04-02
Available Online: 2024-11-23

Abstract

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.
- Intervertebral disc degeneration,
- Human nucleus pulposus cells,
- miR-153-3p,
- Nrf2

FullText(HTML)

References(25)

References

[1]	张嘉军,刘勇,许德荣,郭建伟,王华聪. 椎间盘退变的机制及其治疗综述[J]. 临床医学进展,2020,10(10):2302-2310.
[2]	王延海,张雷明,冯艳艳. 大黄素改善高糖条件中人肾小球血管内皮细胞炎症、氧化应激及凋亡作用的研究[J]. 中国临床药理学杂,2023,39(10):1422-1426.
[3]	杨召,苑珍珍. 炎性因子在中医药治疗椎间盘退变中的机制研究[J]. 中国矫形外科杂志,2021,29(24):2246-2248.
[4]	李敬超. 红景天苷通过Nrf2/ARE信号通路缓解椎间盘退变的研究 [D]. 天津:天津医科大学,2020.
[5]	寇裕,顾勇,陈亮. 黑磷调控氧化应激-炎症级联效应延缓椎间盘退变的机制[J]. 中国组织工程研究,2024,28(15):2338-2345.
[6]	庾珊,肖林,龚东平,等. miR-153-3p对于腰退行性病变调节机制的初步探讨[J]. 昆明医科大学学报,2023,44(8):53-58.
[7]	Ebrahimi S O,Reiisi S,Shareef S. miRNAs,oxidative stress,and cancer: A comprehensive and updated review.[J]. Journal of Cellular Physiology,2020,235(11):8812-8825. doi: 10.1002/jcp.29724
[8]	郭静,吴琦,蔡旖斐,等. miR-153-3p调控神经突的生长[J]. 中华实用诊断与治疗杂志,2019,33(7):650-653.
[9]	张丽霞,周露玙,许胜,等. MiR-153-3p调控异丙肾上腺素诱导的心肌肥大机制研究[J]. 转化医学杂志,2019,8(4):199-202+212.
[10]	崔文洁,袁杰清,施海,等. lncRNA KTN1-AS1调节miR-153-3p/NFAT5轴对非小细胞肺癌细胞增殖、迁移和侵袭的影响[J]. 中国比较医学杂志,2023,33(2):22-30.
[11]	张曹,何亚琴,钱海权,等. miR-153-3p通过靶向FZD3调控胃癌SGC7901细胞的增殖、侵袭与迁移[J]. 中国肿瘤生物治疗杂志,2021,28(9):885-892.
[12]	孙蕾,王华,姜珏,等. miR-153-3p靶向下调Rho相关螺旋蛋白激酶1(ROCK1)基因抑制乳腺癌细胞增殖及迁移[J]. 细胞与分子免疫学杂志,2020,36(2):138-144.
[13]	黄兆琦,许卫,黄炯华,等. 下调miR-153-3p可促进Nrf2表达从而减轻H2O2诱导的H9C2细胞损伤[J]. 中国病理生理杂志,2020,36(7):1249-1254.
[14]	Zou X,Zhang X,Han S,et al. Pathogenesis and therapeutic implications of matrix metalloproteinases in intervertebral disc degeneration: A comprehensive review[J]. Biochimie,2023,214(PtB):27-48.
[15]	周树良,徐良,钱学峰,等. 椎间盘退变程度与髓核中miRNA-142-3p、混合谱系激酶3及白细胞介素1β的相关性[J]. 中国组织工程研究,2024,28(2):165-171.
[16]	D'Arcy M S. Cell death: a review of the major forms of apoptosis,necrosis and autophagy[J]. Cell Biology International,2019,43(6):582-592. doi: 10.1002/cbin.11137
[17]	庞延,卢健棋,唐梅玲,等. 强心汤对慢性心力衰竭模型大鼠梗死心肌区组织活性氧及PINK1/parkin介导的线粒体自噬的影响[J]. 中医杂志,2023,64(7):722-728.
[18]	Picca A,Faitg J,Auwerx J,et al. Mitophagy in human health,ageing and disease.[J]. Nature Metabolism,2023,5(12):2047-2061. doi: 10.1038/s42255-023-00930-8
[19]	潘世鸿,刘瑞端. 线粒体自噬与椎间盘退变[J]. 中国组织工程研究,2024,28(36):5872-5876.
[20]	Quinn P M J,Moreira P I,Ambrosio A F,et al. PINK1/PARKIN signalling in neurodegeneration and neuroinflammation[J]. Acta Neuropathologica Communications,2020,8(1):189. doi: 10.1186/s40478-020-01062-w
[21]	Jahanian S,Pareja-Cajiao M,Gransee H M,et al. Autophagy markers LC3 and p62 in aging lumbar motor neurons[J]. Experimental Gerontology,2024,194:112483.
[22]	魏思灿,林天来,黄玲,等. 槲皮素通过PINK1/parkin通路激活线粒体自噬减轻大鼠脑缺血再灌注损伤[J]. 中国病理生理杂志,2020,36(12):2251-2257.
[23]	Song C,Zhang A,Zhang M,et al. Nrf2/PINK1-mediated mitophagy induction alleviates sodium fluoride-induced hepatic injury by improving mitochondrial function,oxidative stress,and inflammation[J]. Ecotoxicology and Environmental Safety,2023,252:114646. doi: 10.1016/j.ecoenv.2023.114646
[24]	Han Y,Li X,Yan M,et al. Oxidative damage induces apoptosis and promotes calcification in disc cartilage endplate cell through ROS/MAPK/NF-κB pathway: Implications for disc degeneration[J]. Biochem Biophys Res Commun,2019,516(3):1026-1032. doi: 10.1016/j.bbrc.2017.03.111
[25]	Shi Z W,Zhu L,Song Z R,et al. Roles of p38 MAPK signalling in intervertebral disc degeneration[J]. Cell Prolif,2023,56(8):e13438.

Relative Articles(20)

Relative Articles

[1]	Xueya CHEN, Jinmei XU, Zhi LI, Yan LIANG, Yufeng YAO, Fengquan HE, Zhiling YAN. The Association of HOXD-AS2 and MIR3142HG Gene Polymorphisms with Cervical Intraepithelial Neoplasia. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20241103
[2]	Yanping LI, Xiaolin¹ DONG, Qingyun LI, Hongmei LI, Huan WEI, Yi ZENG. miR-21-5p Alleviates OGD/R-induced Inflammation and Apoptosis of HT22 Cells and Promotes Proliferation by Inhibiting STAT3. Journal of Kunming Medical University,
[3]	Xiaobing GUO, Xiaowen LI, Hengxi LI, Yan CAO, Ping LI. miR-212-3p Targeted Regulation of NAP1L1 Inhibits Glioma Cell Proliferation，Migration and EMT. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20241104
[4]	Liang ZHANG, Baoquan WANG, Xifeng LEI, Xu WANG, Yang KE, Wei ZHANG. Effect of miR-29c-3p/IGF1 Molecular Axis on Activation，Proliferation and Apoptosis of Hepatic Stellate Cells. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20230926
[5]	Dong WANG, Bibo GAO, Huiying SUN, Denghui LENG, Xiaoping RAN, Wen LIN. miR-216b-5p Promotes Ferroptosis in Glioblastoma Cells by Targeting NCOA3. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20230805
[6]	Qian XU, Yumei CUI, Siming MA, Yunhong LIN, Yijing XIONG, Zijun SONG, Xudong LI. miR-148a-3p Targeting SMURF2 in Regulating Osteogenic Differentiation and Enamel Development during In Vitro Tooth Organogenesis. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20231103
[7]	Shan YU, Lin XIAO, Dongping GONG, Loufeng LIANG, Anwei LIANG, Xiayi XU, Jie LIN, Huaxin LIANG. The Regulatory Mechanism of miR-153-3p on Lumbar Degeneration. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20230807
[8]	Bin ZHAO, Yuanpeng DUAN, Guoying ZHANG, Chengwei BI, Libo YANG, Zhiyu SHI, Yong YANG, Jianpeng ZHANG, Ting GAO. CircRNA EZH2 Promotes Proliferation and Migration of Prostate Cancer Cells by Regulating miR-30c-5p. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20220731
[9]	Wei ZHANG, Baoquan WANG, Xifeng LEI, Xu WANG, Liang ZHANG. miR-125b-5p Regulates HK2 to Inhibit Proliferation and Glycolysis of Gallbladder Cancer Cells. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20221206
[10]	Xiaobing TAN, Min ZHANG, Yu GUO, Linling DU, Qingyuan DAI. Effect of miR-130b-3p on the Reprogramming of Human Dental Origin iPSCs. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20221204
[11]	Yanlong LIU, Xuefeng GUANG, Xiaolong YIN, Hailong DAI. Effects of miR-125a-3p on Atherosclerotic Plaque, M1/M2 Macrophages, MMP-9 and VEGF. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20220915
[12]	Jing-hui LI, Ming ZHU, Hai QU, Qiu-yu LI, Yu-juan WU, He-ying YANG, Yu-zhu WANG, Yan-ping LI. miR-490-3p Suppresses EMT of SW1990 Pancreatic Cancer Cells. Journal of Kunming Medical University, doi: 10.12259/j.issn.2095-610X.S20210305
[13]	Dai Hai Long , Zhang Xing , Pan Yun , Feng Xiao Lan , Lu Xiang Ting , Guang Xue Feng , Yin Xiao Long . . Journal of Kunming Medical University,
[14]	Li Qin Xue , Li Chuan Jing , Wang Feng . . Journal of Kunming Medical University,
[15]	Bian Li Gong , Zhong Lian Mei , Ai Qing Long , Chen Xin Yue , Xu Wen Kai , Yan Run Qi , Qiu Jin , Lu Di . The Anti-antioxidant Role of Ginsenoside Rg1 Regulating Nrf2 on Focal Cerebral Ischemia Reperfusion SD Rats. Journal of Kunming Medical University,
[16]	Gao Hong Qiang , Liu Jing , Li Zhi Qiang , Wang Hai Lei , Chen Gang , Li Li . . Journal of Kunming Medical University,
[17]	Na Xin , Chen Li Jun , Luo Min , Chen Ya Juan , Qing Chen . . Journal of Kunming Medical University,
[18]	Du Juan . . Journal of Kunming Medical University,
[19]	Li Shan Shan . . Journal of Kunming Medical University,
[20]	Song Chao . . Journal of Kunming Medical University,