Yue Lei, Hu Yongkai, Fu Haoyong, Qi Longtao, Sun Haolin
Department of Orthopaedics Peking University First Hospital, Peking University Beijing China.
Department of Orthopedics General Hospital of Southern Theatre Command of PLA Guangzhou China.
JOR Spine. 2021 Dec 8;4(4):e1181. doi: 10.1002/jsp2.1181. eCollection 2021 Dec.
Hydrogen sulfide (HS) has been found to act as an important gasotransmitter to regulate cell activities. This study aimed to investigate the effect of HS on autophagy of nucleus pulposus (NP) cells under hypoxia and possible mechanism.
NP cells were isolated from rat caudal discs. Cobalt chloride was used to mimic hypoxia, sodium hydrosulfide was used to emulate exogenous HS and 3-methyladenine was used to block cell autophagy. Cell viability was assessed by phase contrast microscope and Cell Counting Kit-8 method. Moreover, expression of key autophagic proteins was analyzed via western blotting, and transmission electron microscopy was performed to detect autophagosomes.
Hypoxia markedly impaired NP cell proliferation compared with control. Whereas HS provided pro-proliferation and pro-autophagy effects on hypoxic NP cells. However, these beneficial impact of HS on hypoxic NP cells were reversed by autophagy inhibitor.
Our results showed that HS played a cytoprotective role in NP cells exposed to hypoxia in an autophagy-dependent manner.
已发现硫化氢（HS）作为一种重要的气体递质来调节细胞活动。本研究旨在探讨HS对缺氧状态下髓核（NP）细胞自噬的影响及其可能机制。
从大鼠尾椎间盘分离出NP细胞。用氯化钴模拟缺氧，用硫氢化钠模拟外源性HS，用3-甲基腺嘌呤阻断细胞自噬。通过相差显微镜和细胞计数试剂盒-8法评估细胞活力。此外，通过蛋白质免疫印迹分析关键自噬蛋白的表达，并进行透射电子显微镜检测自噬体。
与对照组相比，缺氧显著损害NP细胞增殖。而HS对缺氧的NP细胞具有促增殖和促自噬作用。然而，HS对缺氧NP细胞的这些有益影响被自噬抑制剂逆转。
我们的结果表明，HS以自噬依赖的方式在暴露于缺氧的NP细胞中发挥细胞保护作用。
