Department of Orthopedics, XinStell Center Hospital, Xinyu, 338000, JiangXi Province, China.
Department of Orthopedics, The Second Affiliated Hospital of Nanchang University, Nanchang, 330006, JiangXi Province, China.
Biochem Biophys Res Commun. 2018 Sep 5;503(2):791-797. doi: 10.1016/j.bbrc.2018.06.077. Epub 2018 Jun 22.
Serum deprivation is a likely contributor to intervertebral disc (IVD) degeneration (IVDD).17β-estradiol (E2) have been noted to protect nucleus pulposus cells (NPCs) against apoptosis. Autophagy and apoptosis play a paramount role in maintaining the homeostasis of IVD. So far, little research has been published on whether autophagy plays a role for the E2 mediated protection of NPCs. The aim of this study is to understand whether autophagy is involved in the protective effect of E2 against serum deprivation-induced cell apoptosis and expression of matrix metalloproteinase (MMP)-3 and MMP-13. mCherry-GFP-LC3-adenovirus transfection is used to monitor autophagy detection. The expression levels of autophagy-related proteins were measured by Western blotting, Apoptosis and MMPs were detected by flow cytometry and Western blotting. Accordingly, Autophagy and apoptosis was detected in NP cells under serum deprivation conditions, the autophagy incidence began to reached a peak value at 48 h, the apoptosis and MMPs incidence began reached a minimum value treat with E2 (10 M). Whereas the combined use of E2 and 3-MA led to a dramatic decrease in autophagy, while aberrantly elevated expression levels of apoptotic and MMPs. These data suggest that serum deprivation-induced apoptosis and MMP-3, MMP-13, which was efficiently suppressed by the E2 through promoting autophagy in rat NPCs.
血清剥夺可能是导致椎间盘(IVD)退变(IVDD)的原因之一。17β-雌二醇(E2)已被证明可以保护髓核细胞(NPC)免受细胞凋亡。自噬和细胞凋亡在维持 IVD 的内稳态中起着至关重要的作用。到目前为止,关于自噬是否在 E2 介导的 NPC 保护中起作用的研究甚少。本研究旨在了解自噬是否参与 E2 对血清剥夺诱导的细胞凋亡和基质金属蛋白酶(MMP)-3 和 MMP-13 表达的保护作用。使用 mCherry-GFP-LC3-腺病毒转染来监测自噬检测。通过 Western blot 测定自噬相关蛋白的表达水平,通过流式细胞术和 Western blot 检测细胞凋亡和 MMPs。因此,在血清剥夺条件下检测 NP 细胞中的自噬和细胞凋亡,自噬发生率在 48 h 时开始达到峰值,E2(10 M)处理时细胞凋亡和 MMPs 发生率开始达到最小值。而 E2 和 3-MA 的联合使用导致自噬明显减少,而凋亡和 MMPs 的表达水平异常升高。这些数据表明,血清剥夺诱导的细胞凋亡和 MMP-3、MMP-13 被 E2 通过促进 NPC 中的自噬有效抑制。