Expression of Proteinase-Activated Receptor-2 in the Intervertebral Disc
Expression of Proteinase-Activated Receptor-2 in the Intervertebral Disc
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DOI：
10.1097/brs.0b013e318195a67d
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发表时间：
2009-03-01
期刊：
影响因子：
3
通讯作者：
Uchida, Atsumasa
中科院分区：
文献类型：
--
作者：
Iida, Ryu;Akeda, Koji;Uchida, Atsumasa
研究设计.大鼠和人类椎间盘（IVD）中蛋白酶激活受体-2（PAR-2）的免疫组织化学和生物化学分析。研究PAR-2在大鼠IVD细胞中的表达和功能，并确定PAR-2是否在人IVD中表达。PAR-2是一种G蛋白偶联受体，有助于调节炎症反应和炎症性疾病（包括关节炎）的病理生理学。PAR-2在IVD中的表达尚未确定。采用免疫组化和westernblot方法检测大鼠IVD细胞和组织中PAR-2的表达。采用单层培养的大鼠纤维环细胞研究PAR-2的体外生物学作用。通过Western blot和实时荧光定量PCR检测PAR-2激活肽（PAR-2（AP））对分解代谢级联反应的影响。采用免疫组织化学方法检测PAR-2在不同退变阶段人IVD组织中的表达。PAR-2在大鼠椎间盘细胞及纤维环和髓核组织中均有表达，IL-1 β可上调PAR-2的表达。PAR-2（AP）显著增加IL-1 β向培养基中的释放。虽然PAR-2（AP）对基质金属蛋白酶-3（MMP-3）和MMP-13 mRNA水平没有直接影响，但PAR-2（AP）治疗显著上调了具有血小板反应蛋白基序-4的解整合素和金属蛋白酶的mRNA水平。同时给予PAR-2（AP）和IL-1 β协同上调去整合素和金属蛋白酶与血小板反应蛋白基序-4，MMP-3和MMP-13的mRNA水平。PAR-2在人IVD组织中有表达。PAR-2表达细胞在IVD退变晚期较退变早期明显增多。我们的研究结果首次证明IVD细胞表达PAR-2。PAR-2的表达受IL-1 β刺激的调节。PAR-2激活加速基质降解酶的表达。PAR-2可能在精氨酸介导的分解代谢级联反应中发挥重要作用，因此可能参与IVD变性。
Study Design. Immunohistochemical and biochemical analyses of proteinase-activated receptor-2 (PAR-2) in rat and human intervertebral discs (IVDs).Objectives. To examine the expression and function of PAR-2 in rat IVD cells, and to determine if PAR-2 is expressed in human IVDs.Summary of Background Data. PAR-2 is a G protein-coupled receptor that contributes to the regulation of inflammatory reactions and the pathophysiology of inflammatory diseases, including arthritis. The expression of PAR-2 in the IVD has not been determined.Methods. PAR-2 expression by rat IVD cells and tissues was examined using immunohistochemistry and western blot. Rat anulus fibrosus cells in monolayer culture were used to examine the biologic role of PAR-2 in vitro. The effect of PAR-2-activating peptide (PAR-2(AP)) on the catabolic cascade was assessed by western blot and real-time PCR. The expression of PAR-2 by human IVD tissues at different stages of degeneration was determined by immunohistochemical analyses.Results. PAR-2 was expressed by rat IVD cells and in both anulus fibrosus and nucleus pulposus tissues, PAR-2 expression was up-regulated by interleukin-1 beta (IL-1 beta). PAR-2(AP) significantly increased the release of IL-1 beta into the medium. Although PAR-2(AP) had no direct effect on matrix metalloproteinase-3 (MMP-3) and MMP-13 mRNA levels, treatment with PAR-2(AP) significantly up-regulated the mRNA levels of a disintegrin and metalloproteinase with thrombospondin motif-4. The simultaneous administration of PAR-2(AP) and IL-1 beta synergistically up-regulated the mRNA levels of a disintegrin and metalloproteinase with thrombospondin motif-4, MMP-3, and MMP-13. The expression of PAR-2 was identified in human IVD tissues. The number of PAR-2-expressing cells was significantly elevated in advanced stages of IVD degeneration compared with those in early stages of degeneration.Conclusion. Our results demonstrate for the first time that IVD cells express PAR-2. The expression of PAR-2 is regulated by IL-1 beta stimulation. PAR-2 activation accelerates the expression of matrix-degrading enzymes. PAR-2 may play an important role in the cytokine-mediated catabolic cascade and consequently may be involved in IVD degeneration.