Notochordal cells protect nucleus pulposus cells from degradation and apoptosis: implications for the mechanisms of intervertebral disc degeneration.
Notochordal cells protect nucleus pulposus cells from degradation and apoptosis: implications for the mechanisms of intervertebral disc degeneration.
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DOI：
10.1186/ar3548
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发表时间：
2011
影响因子：
4.9
通讯作者：
Tsui FW
中科院分区：
文献类型：
--
作者：
Erwin WM;Islam D;Inman RD;Fehlings MG;Tsui FW
非软骨营养不良（NCD）犬对退行性椎间盘疾病（DDD）的相对抵抗力可能是由于椎间盘（IVD）髓核（NP）内脊索细胞分泌的合成代谢和抗分解代谢因子的组合。已知诱导DDD的因素包括白细胞介素-1 β（IL-1 β）和/或Fas-配体（Fas-L）。因此，我们评估了脊索细胞条件培养基（NCCM）保护NP细胞免受IL-1 β和IL-1 β + FasL介导的细胞死亡和变性的能力。我们在低氧无血清条件下（3.5%O2）用IL-1 β或IL-1 β +FasL培养牛NP细胞，并用无血清NCCM或基础培养基（Advanced DMEM/F-12）处理细胞。我们使用流式细胞术来评估细胞死亡和实时（RT-）PCR来确定聚集蛋白聚糖，胶原蛋白2和连接蛋白，基质降解介质ADAMTS-4和MMP 3，基质保护分子TIMP 1，分化簇（CD）44受体，炎性细胞因子IL-6和Ank的基因表达。然后，我们通过表征当与NCCM、使用牛NP细胞获得的条件培养基（BCCM）和全部补充有2%FBS的基础培养基一起培养时，在IL-1 β +FasL存在下活化的半胱天冬酶-3、-8和-9的表达，确定牛NP细胞中特异性凋亡途径的表达。NCCM通过抑制活化的caspase-9和caspase-3/7抑制牛NP细胞死亡和凋亡。此外，NCCM通过上调合成代谢/基质保护基因（聚集蛋白聚糖、2型胶原、CD 44、连接蛋白和TIMP-1）的表达和下调基质降解基因（如MMP-3），保护NP细胞免受IL-1 β和IL-1 β +Fas-L的降解作用。ADAMTS-4的表达增加，其编码聚集蛋白聚糖重塑的蛋白。NCCM还防止IL-1+ FasL介导的Ank表达下调。此外，在IL-1 β +Fas-L存在下用NCCM处理的NP细胞下调IL-6的表达几乎50%。BCCM不介导靶牛NP细胞中的细胞死亡/凋亡。脊索细胞分泌的因子通过抑制活化的半胱天冬酶-9和-3/7活性以及通过上调促进IVD NP的合成代谢活性和基质保护的基因来抑制NP细胞死亡。利用脊索细胞的恢复能力可能会导致新的细胞和分子策略治疗DDD。
The relative resistance of non-chondrodystrophic (NCD) canines to degenerative disc disease (DDD) may be due to a combination of anabolic and anti-catabolic factors secreted by notochordal cells within the intervertebral disc (IVD) nucleus pulposus (NP). Factors known to induce DDD include interleukin-1 beta (IL-1ß) and/or Fas-Ligand (Fas-L). Therefore we evaluated the ability of notochordal cell conditioned medium (NCCM) to protect NP cells from IL-1ß and IL-1ß +FasL-mediated cell death and degeneration. We cultured bovine NP cells with IL-1ß or IL-1ß+FasL under hypoxic serum-free conditions (3.5% O2) and treated the cells with either serum-free NCCM or basal medium (Advanced DMEM/F-12). We used flow cytometry to evaluate cell death and real-time (RT-)PCR to determine the gene expression of aggrecan, collagen 2, and link protein, mediators of matrix degradation ADAMTS-4 and MMP3, the matrix protection molecule TIMP1, the cluster of differentiation (CD)44 receptor, the inflammatory cytokine IL-6 and Ank. We then determined the expression of specific apoptotic pathways in bovine NP cells by characterizing the expression of activated caspases-3, -8 and -9 in the presence of IL-1ß+FasL when cultured with NCCM, conditioned medium obtained using bovine NP cells (BCCM), and basal medium all supplemented with 2% FBS. NCCM inhibits bovine NP cell death and apoptosis via suppression of activated caspase-9 and caspase-3/7. Furthermore, NCCM protects NP cells from the degradative effects of IL-1ß and IL-1ß+Fas-L by up-regulating the expression of anabolic/matrix protective genes (aggrecan, collagen type 2, CD44, link protein and TIMP-1) and down-regulating matrix degrading genes such as MMP-3. Expression of ADAMTS-4, which encodes a protein for aggrecan remodeling, is increased. NCCM also protects against IL-1+FasL-mediated down-regulation of Ank expression. Furthermore, NP cells treated with NCCM in the presence of IL-1ß+Fas-L down-regulate the expression of IL-6 by almost 50%. BCCM does not mediate cell death/apoptosis in target bovine NP cells. Notochordal cell-secreted factors suppress NP cell death by inhibition of activated caspase-9 and -3/7 activity and by up-regulating genes contributing anabolic activity and matrix protection of the IVD NP. Harnessing the restorative powers of the notochordal cell could lead to novel cellular and molecular strategies in the treatment of DDD.
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影响因子：
4.9
作者：
Le Maitre, Christine Lyn;Hoyland, Judith Alison;Freemont, Anthony J
通讯作者：
Freemont, Anthony J
影响因子：
5.3
作者：
Park, JB;Li, JK;Riew, KD
通讯作者：
Riew, KD
影响因子：
--
作者：
Erwin, W. Mark;Ashman, Keith;Inman, Robert D.
通讯作者：
Inman, Robert D.
影响因子：
5.5
作者：
Hoyland, J. A.;Le Maitre, C.;Freemont, A. J.
通讯作者：
Freemont, A. J.
DOI：
10.1152/ajpgi.00160.2004
发表时间：
2005-02-01
影响因子：
4.5
作者：
Nomura, S;Yamaguchi, H;Goldenring, JR
通讯作者：
Goldenring, JR