Yang Xuefang
Orthopedics Department, Yingkou Central Hospital, Yingkou, China.
Ann Transl Med. 2022 Feb;10(4):213. doi: 10.21037/atm-22-91.
The intervertebral disc can increase the amplitude of spinal motion, withstand pressure, buffer vibration, and protect the brain and spinal cord. It is also the main reason why height changes, but the regulatory mechanism is still unclear, and this study mainly explored the role of miR-874-3p in intervertebral disc degeneration (IDD).
The mechanism and perform correlation analysis of miR-874-3p and the pathological degree and prognosis of patients with IDD. miR-874-3p is involved in the progression of several diseases, such as cell differentiation, proliferation, apoptosis and extracellular matrix degradation, overexpressing cell line GV369-miR-874-3p-NP was obtained by infected nucleus pulposus (NP) cells, and the empty vector GV369-NP group was set with the blank group. The expression of the tag protein (green fluorescent protein, GFP) was visualized by fluorescence microscopy, followed by real-time polymerase chain reaction (PCR) method to detect miR-874-3p expression, apoptosis by flow cytometry, luciferase reporter analysis for verifying the targeting relationship between miR-874-3p, caspase-3, B-cell lymphoma-2 (Bcl-2) and Bax in cells, and examined the changes in cellular mitochondrial membrane potential using kits.
The expression of miR-874-3p was significantly reduced in IDD patients, and was negatively correlated with matrix metallopeptidase 2 (MMP2) and downregulated matrix metallopeptidase 3 (MMP3) in the NP cells. In addition, western blot revealed that overexpression of miR-874-3p increased the aggregation protein level in the NP cells.
miR-874-3p can inhibit the cell death of IDD, and not only participate in caspase-3 and Fas-associating protein with a novel death domain (FADD)-mediated apoptosis through targeted regulation of exogenous MMP2/MMP3 pathway, but also play a role in cell apoptosis through mitochondrial pathway.
椎间盘可增加脊柱运动幅度、承受压力、缓冲震动并保护脑和脊髓。它也是身高变化的主要原因,但其调节机制仍不清楚,本研究主要探讨了miR-874-3p在椎间盘退变(IDD)中的作用。
对miR-874-3p与IDD患者病理程度及预后进行机制及相关性分析。miR-874-3p参与多种疾病的进展,如细胞分化、增殖、凋亡及细胞外基质降解,通过感染髓核(NP)细胞获得过表达细胞系GV369-miR-874-3p-NP,并设置空载体GV369-NP组及空白组。通过荧光显微镜观察标签蛋白(绿色荧光蛋白,GFP)的表达,随后采用实时聚合酶链反应(PCR)法检测miR-874-3p表达,通过流式细胞术检测凋亡情况,采用荧光素酶报告基因分析验证细胞中miR-874-3p、半胱天冬酶-3(caspase-3)、B细胞淋巴瘤-2(Bcl-2)和Bax之间的靶向关系,并使用试剂盒检测细胞线粒体膜电位的变化。
IDD患者中miR-874-3p表达显著降低,且与NP细胞中基质金属蛋白酶2(MMP2)及下调的基质金属蛋白酶3(MMP3)呈负相关。此外,蛋白质免疫印迹法显示miR-874-3p过表达增加了NP细胞中的聚集蛋白水平。
miR-874-3p可抑制IDD的细胞死亡,不仅通过靶向调控外源性MMP2/MMP3途径参与caspase-3和含新死亡结构域的Fas相关蛋白(FADD)介导的凋亡,还通过线粒体途径在细胞凋亡中发挥作用。