YAP通过促进自噬抑制髓核细胞胞外基质分解代谢

易威威; 唐秋雨; 陶白龙; 李开庭; 王大武

doi:10.16016/j.2097-0927.202310058

您当前的位置：

首页 >

文章列表页 >

YAP通过促进自噬抑制髓核细胞胞外基质分解代谢

基础医学 | 更新时间：2026-01-09

- YAP通过促进自噬抑制髓核细胞胞外基质分解代谢
- YAP inhibits extracellular matrix catabolism of nucleus pulposus cells by promoting autophagy
- 陆军军医大学学报 2024年46卷第10期页码：1107-1114
- 作者机构：
  
  1.400016 重庆，重庆医科大学附属第一医院: 康复医学科
  2.400016 重庆，重庆医科大学附属第一医院: 实验研究中心
- 作者简介：
  
  [王大武，E-mail: wangdawu66@163.com WANG Dawu, E-mail: wangdawu66@163.com
- 基金信息：
  
  国家自然科学基金面上项目(82102537);重庆市自然科学基金(CSTB2022NSCQ-MSX0064);重庆市自然科学基金(CSTB2022BSXM-JCX0058)
- DOI：10.16016/j.2097-0927.202310058
  中图分类号： R341;R363.21;R681.53
- 收稿：2023-10-13，
  
  修回：2023-11-26，
  
  纸质出版：2024-05-30
- 稿件说明：
移动端阅览
易威威, 唐秋雨, 陶白龙, 等. YAP通过促进自噬抑制髓核细胞胞外基质分解代谢[J]. 陆军军医大学学报, 2024,46(10):1107-1114.

Weiwei YI, Qiuyu TANG, Bailong TAO, et al. YAP inhibits extracellular matrix catabolism of nucleus pulposus cells by promoting autophagy[J]. Journal of Army Medical University, 2024, 46(10): 1107-1114.
易威威, 唐秋雨, 陶白龙, 等. YAP通过促进自噬抑制髓核细胞胞外基质分解代谢[J]. 陆军军医大学学报, 2024,46(10):1107-1114. DOI： 10.16016/j.2097-0927.202310058.

Weiwei YI, Qiuyu TANG, Bailong TAO, et al. YAP inhibits extracellular matrix catabolism of nucleus pulposus cells by promoting autophagy[J]. Journal of Army Medical University, 2024, 46(10): 1107-1114. DOI： 10.16016/j.2097-0927.202310058.

摘要

目的

探讨YAP对椎间盘髓核细胞的作用及其可能机制。

方法

收集重庆医科大学附属第一医院2021年3月至2022年7月接受腰椎手术患者的相对正常和退变的椎间盘组织，采用免疫组化和Western blot检测YAP在人椎间盘髓核组织中的表达差异。体外培养人原代髓核细胞，通过IL-1β处理诱导髓核细胞退变，将实验分为对照组、IL-1β组、IL-1β+LV-YAP组、IL-1β+YAP-siRNA组和IL-1β+LV-YAP+3-MA组。采用Western blot检测细胞外基质分解代谢及自噬水平变化。最后建立大鼠椎间盘退变模型，利用MRI、阿利新蓝染色及免疫组化检测YAP和LC3的表达及椎间盘退变情况。

结果

YAP在退变椎间盘组织的表达水平明显低于相对正常的椎间盘组织(

0.05)。体外细胞实验结果显示，IL-1β+LV-YAP组显著提高IL-1β诱导的髓核细胞中Collagen Ⅱ、Aggrecan和LC3-Ⅱ的蛋白表达(

0.05)，降低MMP-3和MMP-13的蛋白表达(

0.05)，而IL-1β+YAP-siRNA组则表现出完全相反的作用。而予以自噬抑制剂3-MA预处理后明显降低IL-1β+LV-YAP组中GFP-LC3阳性颗粒数量和Collagen Ⅱ、Aggrecan、LC3-Ⅱ蛋白表达(

0.05)，且提高MMP-3和MMP-13的蛋白表达(

0.05)。进一步在体内构建大鼠椎

间盘退变模型，YAP过表达促进LC3表达和抑制椎间盘退变。

结论

YAP过表达通过促进人髓核细胞自噬抑制细胞外基质降解，从而延缓椎间盘退变。

Abstract

Objective

To investigate the effect of Yes-associated protein (YAP) on intervertebral disc nucleus pulposus cells and its possible mechanism.

Methods

The relatively normal and degenerative intervertebral disc tissues of patients who underwent lumbar surgery in our hospital from March 2021 to July 2022 were harvested

and then the expression of YAP in the tissues were detected by immunohistochemistry assay and Western blotting. Human primary nucleus pulposus cells were isolated and primarily cultured

and treated with IL-1β to induce degeneration. Then the cells was divided into control group

IL-1β group

IL-1β+LV-YAP group

IL-1β+YAP-siRNA group

and IL-1β+LV-YAP+3-MA group. Western blot analysis was used to detect the expression of the proteins related to extracellular matrix catabolism and autophagy in each group. Finally

a rat model of disc degeneration was established

and the expression of YAP and LC3 and disc degeneration were observed with MRI

Alcian blue staining and immunohistochemistry.

Results

The expression level of YAP was significantly lower in the degraded disc tissues than the relatively normal disc tissues (

0.05). The IL-1β+LV-YAP group had significantly increased protein levels of Collagen Ⅱ

Aggrecan

and LC3-Ⅱ (

0.05)

and decreased levels of MMP-3 and MMP-13 (

0.05) when compared with the cells after IL-1β treatment

whereas the IL-1β+YAP-siRNA group showed the exact opposite effects. What's more

pretreatment with autophagy inhibitor 3-MA resulted in decreased number of GFP-LC3 positive particles and protein levels of Collagen Ⅱ

Aggrecan and LC3-Ⅱ (

0.05)

and increased protein expression of MMP-3 and MMP-13 (

0.05) in comparison with the conditions in the IL-1β+LV-YAP group. Furthermore

YAP overexpression promoted LC3 expression and inhibited disc degeneration in rat model of disc degeneration.

Conclusion

YAP overexpression can inhibit extracellular matrix degradation by promoting autophagy in human nucleus pulposus cells and thus delaying disc degeneration.

关键词

Keywords

references

WU T D, JIA X B, FENG H, et al . ACTG1 regulates intervertebral disc degeneration via the NF-κB-p65 and Akt pathways[J ] . Biochem Biophys Res Commun, 2021, 545: 54-61. DOI: 10.1016/j.bbrc.2021.01.057..

ZHANG C, WANG F, XIE Z Y, et al . AMOT130 linking F-actin to YAP is involved in intervertebral disc degeneration[J ] . Cell Prolif, 2018, 51(6): e12492. DOI: 10.1111/cpr.12492.

PICCOLO S, DUPONT S, CORDENONSI M. The biology of YAP/TAZ: hippo signaling and beyond[J]. Physiol Rev, 2014, 94(4): 1287-1312. DOI: 10.1152/physrev.00005.2014.

PEI T J, HUANG X, LONG Y, et al . Increased expression of YAP is associated with decreased cell autophagy in the eutopic endometrial stromal cells of endometriosis[J ] . Mol Cell Endocrinol, 2019, 491: 110432. DOI: 10.1016/j.mce.2019.04.012.

DENG Y J, LU J Q, LI W L, et al . Reciprocal inhibition of YAP/TAZ and NF-κB regulates osteoarthritic cartilage degradation[J ] . Nat Commun, 2018, 9(1): 4564. DOI: 10.1038/s41467-018-07022-2.

GE J, YANG H L, CHEN Y F, et al . PMMA bone cement acts on the hippo/YAP pathway to regulate CTGF and induce intervertebral disc degeneration[J ] . ACS Biomater Sci Eng, 2019, 5(7): 3293-3302. DOI: 10.1021/acsbiomaterials.9b00146.

GONG C Y, ZHANG H H. Autophagy as a potential therapeutic target in intervertebral disc degeneration[J]. Life Sci, 2021, 273: 119266. DOI: 10.1016/j.lfs.2021.119266.

ZHANG S J, YANG W, WANG C, et al . Autophagy: a double-edged sword in intervertebral disk degeneration[J ] . Clin Chimica Acta, 2016, 457: 27-35. DOI: 10.1016/j.cca.2016.03.016.

ZHANG Y, HE F, CHEN Z, et al . Melatonin modulates IL-1β-induced extracellular matrix remodeling in human nucleus pulposus cells and attenuates rat intervertebral disc degeneration and inflammation[J ] . Aging (Albany NY), 2019, 11(22): 10499-10512. DOI: 10.18632/aging.102472.

CHEN J, MA Y F, YANG Z J, et al . TNFAIP3 ameliorates the degeneration of inflammatory human nucleus pulposus cells by inhibiting mTOR signaling and promoting autophagy[J ] . Aging (Albany NY), 2020, 12(23): 24242-24254. DOI: 10.18632/aging.104160.

WANG D Y, HE J X, HUANG B Y, et al . Emerging role of the Hippo pathway in autophagy[J ] . Cell Death Dis, 2020, 11(10): 880. DOI: 10.1038/s41419-020-03069-6.

ZHOU W, SHEN Q, WANG H, et al . Knockdo wn of YAP/TAZ inhibits the migration and invasion of fibroblast synovial cells in rheumatoid arthritis by regulating autophagy[J ] . J Immunol Res, 2020, 2020: 9510594. DOI: 10.1155/2020/9510594.

PFIRRMANN C W A, METZDORF A, ZANETTI M, et al . Magnetic resonance classification of lumbar intervertebral disc degeneration[J ] . Spine, 2001, 26(17): 1873-1878. DOI: 10.1097/00007632-200109010-00011.

DOWDELL J, ERWIN M, CHOMA T, et al . Intervertebral disk degeneration and repair[J ] . Neurosurgery, 2017, 80(3S): S46-S54. DOI: 10.1093/neuros/nyw078.

YANG S D, ZHANG F, MA J T, et al . Intervertebral disc ageing and degeneration: the antiapoptotic effect of oestrogen[J ] . Ageing Res Rev, 2020, 57: 100978. DOI: 10.1016/j.arr.2019.100978.

ZHANG X B, HU Y C, CHENG P, et al . Targeted therapy for intervertebral disc degeneration: inhibiting apoptosis is a promising treatment strategy[J ] . Int J Med Sci, 2021, 18(13): 2799-2813. DOI: 10.7150/ijms.59171.

PUTS R, RIKEIT P, RUSCHKE K, et al . Functional regulation of YAP mechanosensitive transcriptional coactiv ator by Focused Low-Intensity Pulsed Ultrasound (FLIPUS) enhances proliferation of murine mesenchymal precursors[J ] . PLoS One, 2018, 13(10): e0206041. DOI: 10.1371/journal.pone.0206041.

ZHANG C, WANG F, XIE Z Y, et al . Dysregulation of YAP by the Hippo pathway is involved in intervertebral disc degeneration, cell contact inhibition, and cell senescence[J ] . Oncotarget, 2018, 9(2): 2175-2192. DOI: 10.18632/oncotarget.23299.

CHEN W, BAI Y X, PATEL C, et al . Autophagy promotes triple negative breast cancer metastasis via YAP nuclear localization[J ] . Biochem Biophys Res Commun, 2019, 520(2): 263-268. DOI: 10.1016/j.bbrc.2019.09.133..

MOYA I M, HALDER G. Hippo-YAP/TAZ signalling in organ regeneration and regenerative medicine[J]. Nat Rev Mol Cell Biol, 2019, 20(4): 211-226. DOI: 10.1038/s41580-018-0086-y.

ZHU J, ZHAO B X, XIONG P G, et al . Curcumin induces autophagy via inhibition of Yes-associated protein (YAP) in human colon cancer cells[J ] . Med Sci Monit, 2018, 24: 7035-7042. DOI: 10.12659/MSM.910650..

WEI L X, CAO P, XU C, et al . Chondroitin synthase-3 regulates nucleus pulposus degeneration through actin-induced YAP signaling[J ] . FASEB J, 2020, 34(12): 16581-16600. DOI: 10.1096/fj.202001021R.

LUO Z Q, MA Y C, DI T N, et al . DNMT3B decreases extracellular matrix degradation and alleviates intervertebral disc degeneration through TRPA1 methylation to inhibit the COX2/YAP axis[J ] . Aging (Albany NY), 2021, 13(16): 20258-20276. DOI: 10.18632/aging.203410.

CORTI O, BLOMGREN K, POLETTI A, et al . Autophagy in neurodegeneration: new insights underpinning therapy for neurological diseases[J ] . J Neurochem, 2020, 154(4): 354-371. DOI: 10.1111/jnc.15002.

DUAN R, XIE H, LIU Z Z. The role of autophagy in osteoarthritis[J]. Front Cell Dev Biol, 2020, 8: 608388. DOI: 10.3389/fcell.2020.608388.

ZHANG T W, LI Z F, DONG J, et al . The circadian rhythm in intervertebral disc degeneration: an autophagy connection[J ] . Exp Mol Med, 2020, 52(1): 31-40. DOI: 10.1038/s12276-019-0372-6.

CHEN F, LIU H, WANG X, et al . Melatonin activates autophagy via the NF-κB signaling pathway to prevent extracellular matrix degeneration in intervertebral disc[J ] . Osteoarthr Cartilage, 2020, 28(8): 1121-1132. DOI: 10.1016/j.joca.2020.05.011..

TOTARO A, ZHUANG Q Y, PANCIERA T, et al . Cell phenotypic plasticity requires autophagic flux driven by YAP/TAZ mechanotransduction[J ] . Proc Natl Acad Sci U S A, 2019, 116(36): 17848-17857. DOI: 10.1073/pnas.1908228116.

ZHAO M, ZHANG Y, JIANG Y, et al . YAP promotes autophagy and progression of gliomas via upregulating HMGB1[J ] . J Exp Clin Cancer Res, 2021, 40(1): 99. DOI: 10.1186/s13046-021-01897-8..

ZHOU Y, WANG Y B, ZHOU W H, et al . YAP promotes multi-drug resistance and inhibits autophagy-related cell death in hepatocellular carcinoma via the RAC1-ROS-mTOR pathway[J ] . Cancer Cell Int, 2019, 19: 179. DOI: 10.1186/s12935-019-0898-7..

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

Sirt1通过调控髓核细胞组蛋白H3K18乳酸化参与椎间盘退变的机制研究

Kallistatin诱导SIRT1表达抑制人髓核细胞凋亡及细胞外基质降解

锌指蛋白A20对内毒素刺激时人椎间盘髓核细胞炎症及退变的影响

缺氧诱导锌指蛋白A20表达抑制髓核细胞凋亡和炎症

RANKL通过ROS介导HIF-1α下调促进自噬和破骨细胞分化