APOPTOSIS OF CELLS OF NUCLEUS PULPOSUS AND ACTIVATION MECHANISM OF INTERCELLULAR SIGNALING DURING DEGENERATION OF INTERVERTEBRAL DISC

椎间盘退变过程中髓核细胞的凋亡及细胞间信号传导的激活机制

• 批准号: 13671523
• 负责人: MATSUNAGA Shunji
• 金额: $ 2.3万
• 依托单位: Kagoshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671523/
• 关键词: senescence-accelerated mice; intervertebral disc; cytokines; apoptosis; intercellular signaling; 変性; TGF-β; 分子生物学; 老化; 髄核細胞; 細胞内シグナル伝達

We previously conducted immunohistological studies of changes in TGF-β and BMP expression in nucleus pulposus cells following degeneration of the intervertebral disc. These studies suggested that changes in TGF-β expression are associated with apoptosis of cells. Following this finding, the present study was undertaken to clarify the mechanism of aging from the viewpoint of apoptosis, using molecular biological methods. The study involved a senescence-accelerated mouse strain (SAM-P/6) as the animal model, allowing observation of aging in a short time period. During last year, we removed the intervertebral discs of the cervical spine from male senescence-accelerated mice at ages 4, 8, 12, 16, 20, 24, 28, 32 and 50 weeks and examined immunohistologically the expression of TGF-β1, -β2, -β3 and their receptors in the discs' nucleus pulposus cells. The study revealed that as age advanced the expression of nucleus pulposus cell TGF-β1, -β2, -β3 and their receptors decreased, reaching almost … More zero at age 50 weeks and was accompanied by a decrease in the number of nucleus pulposus cells. This year, we conducted an experiment designed to demonstrate that such a decrease in nucleus pulposus cells is due to apoptosis. In this study, nucleus pulp cells of senescence-accelerated mice were stained positively using the TUNEL method and were found to already have caspase activity at age 32 weeks, suggesting that apoptosis had begun at this age. Morphological observation of the nuclei of the cells with an electron microscope also revealed apoptotic changes. The changes suggesting apoptosis were more marked at age 50 weeks.In 2003, we conducted immunohistological studies of ASK1, JNK and p38 aimed at clarifying the mechanism for intracellular signal transduction associated with apoptosis during degeneration of intervertebral discs. As the disc degenerated, the expression of ASK1, JNK and p38 in nucleus pulposus cells was noted, but it was also observed in the control group. Thus, it was not possible to clarify the mechanism for intracellular signal transduction associated with apoptosis. Less

我们先前进行了椎间盘退变后髓核细胞中TGF-β和BMP表达变化的免疫组织学研究。这些研究表明，TGF-β表达的变化与细胞凋亡有关。根据这一发现，本研究从细胞凋亡的角度，用分子生物学方法阐明衰老的机制。该研究涉及衰老加速小鼠品系（SAM-P/6）作为动物模型，允许在短时间内观察衰老。本研究采用4、8、12、16、20、24、28、32、50周龄的快速老化雄性小鼠，取颈椎间盘，用免疫组织化学方法检测TGF-β1、β2、β3及其受体在椎间盘髓核细胞中的表达。研究发现，随着年龄的增长，髓核细胞TGF-β1、TGF-β2、TGF-β3及其受体的表达逐渐减少， ...更多信息 在50周龄时为零，并伴有髓核细胞数量的减少。今年，我们进行了一项实验，旨在证明髓核细胞的减少是由于细胞凋亡造成的。在这项研究中，使用TUNEL方法对加速衰老小鼠的髓核细胞进行了阳性染色，发现在32周龄时已经具有caspase活性，这表明细胞凋亡在这个年龄已经开始。电镜观察细胞核形态也显示凋亡改变。2003年，我们对ASK 1、JNK和p38进行了免疫组织化学研究，旨在阐明椎间盘退变过程中细胞内信号转导与凋亡相关的机制。随着椎间盘退变，髓核细胞中有ASK 1、JNK和p38的表达，而对照组中也有表达。因此，不可能阐明与细胞凋亡相关的细胞内信号转导机制。少

项目成果

Nagano S, Matsunaga S, Yamamoto T, Yone K, Sakou, Komiya S: "Immunolocalization of transforming growth factor-betas and their receptors in the intervertebral disk of cervical spine"Proceedings of Fourth Combined Meeting of the Orthopawedic research Societ

Nagano S、Matsunaga S、Yamamoto T、Yone K、Sakou、Komiya S：“颈椎椎间盘中转化生长因子-β及其受体的免疫定位”骨科研究会第四次联合会议论文集

I.Yamaura, K.Yone, et al.: "Mechanism of destractive pathologic changes in spinal cord under clonic mechanical compression."Spine. 27. 21-26 (2002)

I.Yamaura、K.Yone 等人：“阵挛机械压迫下脊髓破坏性病理变化的机制。”脊柱。

S.Matsunaga, S.Nagano, T.Onishi, N.Morimoto, S.Suzuki, S.Komiya: "Age-related changes in expression of transforming growth factor-β and receptors in cells of intervertebral discs."J Neurosurg. 98. 63-67 (2003)

S.Matsunaga、S.Nagano、T.Onishi、N.Morimoto、S.Suzuki、S.Komiya：“椎间盘细胞中转化生长因子-β 和受体表达的年龄相关变化。”J Neurosurg。 .63-67 (2003)

S.Matsunaga, S.Nagano, et al.: "Age-related chages in expression of transforming growth factor-β and receptors in cells of intervertebral discs."J.Neurosurg.. 98. 63-67 (2003)

S.Matsunaga、S.Nagano 等人：“椎间盘细胞中转化生长因子-β 和受体表达的年龄相关变化。J.Neurosurg.. 98. 63-67 (2003)

Shunji Matsunaga: "Age-related changes in expression of transforming growth factor-β and receptors in cells of intervertebral discs"J Neurosurg. 98. 63-67 (2003)

Shunji Matsunaga：“椎间盘细胞中转化生长因子-β 和受体表达的年龄相关变化”J Neurosurg。 98. 63-67 (2003)