Tension induces intervertebral disc degeneration via endoplasmic reticulum stress-mediated autophagy
Tension induces intervertebral disc degeneration via endoplasmic reticulum stress-mediated autophagy
复制标题
张力通过内质网应激介导的自噬诱导椎间盘退变
DOI：
10.1042/bsr20190578
复制
发表时间：
2019-08-07
影响因子：
4
通讯作者：
Yang, Dong
中科院分区：
文献类型：
--
作者：
Chen, Jiangwei;Lin, Zunwen;Yang, Dong
背景：椎间盘退变是一种常见的退行性疾病。本研究旨在探讨张力诱导的内质网应激在椎间盘退变中的作用及其机制。研究方法：采用免疫组化、qPCR和Western blot方法检测SD大鼠椎间盘退变模型中细胞凋亡、多聚腺苷二磷酸核糖聚合酶（PARP）、Caspase-12、Caspase-3、LC 3、Beclin-1和CHOP的表达。分离椎间盘纤维环细胞，进行周期性变形应力，并分析ROS和凋亡，溶酶体活性和基因表达。用siRNA敲低细胞或用内质网应激抑制剂4-PBA处理细胞，并测定ROS、凋亡、溶酶体活性和基因表达。结果：与对照组相比，X线片和HS染色显示椎间盘退变。模型组椎间盘组织中PARP、Caspase-12、Caspase-3、LC 3、Beclin-1和CHOP的表达及细胞凋亡均明显增加。在力学模拟实验中，原代纤维环细胞经受18%的周期性形变，细胞内活性氧和细胞凋亡增加，溶酶体活性增加。同样，PARP、Caspase-12、Caspase-3、LC 3、Beclin-1和CHOP的表达在变形处理后也显著增加。另一方面，当用9 mM 4-PBA和/或CHOP-siRNA 4处理细胞时，凋亡率、ROS水平、溶酶体活性和PARP、Caspase-12、Caspase-3、LC 3、Beclin-1和CHOP的表达显著降低。结论：内质网应激介导的自噬反应在张力性椎间盘退变中起重要作用。椎间盘退变可能是自噬、凋亡和网状应激之间相互作用的结果，并且是ROS依赖的。
Background: Intervertebral disc degeneration is a common degenerative disease. The present study aimed to explore the role and mechanism of tension-induced endoplasmic reticulum stress in intervertebral disc degeneration. Methods: Intervertebral disc degeneration models of SD rat were analyzed for apoptosis, the expression of Poly(ADP-ribose) polymerase (PARP), Caspase-12, Caspase-3, LC3, Beclin-1 and CHOP using immunohistochemistry, qPCR and Western blot analysis. Annulus fibrosus cells of intervertebral disc were isolated, subjected to cyclic deformation stress and analyzed for ROS and apoptosis, lysosome activity and expression of genes. The cells were knockdown with siRNA or treated with endoplasmic reticulum stress inhibitor 4-PBA and assayed for ROS, apoptosis, lysosome activity and gene expression. Results: Compared with the controls, intervertebral disc degeneration was observed through X-rays examinations and HS staining. Apoptosis and expression of PARP, Caspase-12, Caspase-3, LC3, Beclin-1 and CHOP were significantly increased in the intervertebral disc tissue of the models. In mechanic mimic experiments, the primary annulus fibrosus cells were subjected to 18% cyclic deformation, ROS and apoptosis as well as the activity of lysosome were increased. Similarly, the expression of PARP, Caspase-12, Caspase-3, LC3, Beclin-1 and CHOP was also increased significantly after deformation treatment. On other hand, when the cells were treated with 9 mM 4-PBA and/or CHOP-siRNA4, the apoptosis rate, ROS level, lysosome activity and expression of PARP, Caspase-12, Caspase-3, LC3, Beclin-1 and CHOP were significantly reduced. Conclusions: Autophagy reaction mediated by endoplasmic reticulum stress plays important rale in tension-induced intervertebral disc degeneration. Intervertebral disc degeneration likely results from interactions between autophagy, apoptosis and reticulum stress, and is ROS-dependent.