Sun Zhongyi, Yin Zhanmin, Liu Chao, Tian Jiwei
Department of Orthopedics Surgery, School of Medicine, Shanghai First People's Hospital Affiliated Shanghai Jiao Tong University, 100, Haining Road, Shanghai, 200080, China.
Spine and Joint Surgery, Central Hospital of Tai An, Shandong, China.
Cell Biochem Biophys. 2015 May;72(1):115-22. doi: 10.1007/s12013-014-0417-3.
We aim at determining the changes in the expression of NF-kB signaling pathway in degenerative intervertebral discs. We collected normal and degenerated intervertebral discs tissues. The normal and degenerated cells were cultivated and their histopathology and immunofluoresence studies were used to observe the position of NF-kB p65 in the cell. We also treated the nucleus pulposus cells with inflammatory factors and inhibitors. Western blot was used to analyze the expression of different proteins. Real time fluorescence-based quantitative PCR was used for observation of NF-kB regulation of change in gene expression. Immunofluorescence showed that in the non-degenerative group the p65 was found in the cytoplasm of the nucleus pulposus cell while in the degenerated cell group the p65 protein was found in the nucleus of the cell. The expression of p65 increased with increase in the degree of degenerative change of the nucleus pulposus cell. RT-PCR showed that the expression of matrix metalloproteinases, aggrecanases and IL-6 was higher in the degenerative group. The amount of aggrecan and type II collagen was significantly decreased in the degenerative group. IL-1β was able to upregulate the activation of NF-kB and the expression of MMP-13 and ADAMTS-4 was also significantly increased. The effect of these proteins can be inhibited by the NF-kB inhibitor, BAY11-7082. The activation of the NK-kB signaling pathway in a degenerative intervertebral disc is gradually increased, regulating the over-expression of matrix-degrading enzymes. It plays an important role in the degradation of extracellular matrix.
我们旨在确定退变椎间盘组织中核因子-κB（NF-κB）信号通路表达的变化。我们收集了正常和退变的椎间盘组织。对正常和退变细胞进行培养，并通过组织病理学和免疫荧光研究观察NF-κB p65在细胞中的位置。我们还用炎性因子和抑制剂处理髓核细胞。采用蛋白质印迹法分析不同蛋白质的表达。采用实时荧光定量PCR观察NF-κB对基因表达变化的调控。免疫荧光显示，在非退变组中，p65存在于髓核细胞的细胞质中，而在退变细胞组中，p65蛋白存在于细胞核中。p65的表达随着髓核细胞退变程度的增加而增加。逆转录聚合酶链反应（RT-PCR）显示，退变组中基质金属蛋白酶、聚集蛋白聚糖酶和白细胞介素-6（IL-6）的表达较高。退变组中聚集蛋白聚糖和Ⅱ型胶原蛋白的含量显著降低。白细胞介素-1β（IL-1β）能够上调NF-κB的激活，基质金属蛋白酶-13（MMP-13）和含血小板反应蛋白基元的解聚蛋白样金属蛋白酶-4（ADAMTS-4）的表达也显著增加。这些蛋白质的作用可被NF-κB抑制剂BAY11-7082抑制。退变椎间盘中NF-κB信号通路的激活逐渐增加，调节基质降解酶的过度表达。它在细胞外基质的降解中起重要作用。
