Ariga K, Yonenobu K, Nakase T, Kaneko M, Okuda S, Uchiyama Y, Yoshikawa H
Department of Orthopaedic Surgery, Osaka University Medical School, Suita, Osaka, Japan.
Spine (Phila Pa 1976). 2001 Dec 15;26(24):2666-72. doi: 10.1097/00007632-200112150-00007.
Localization of cathepsins D, K, and L in degenerated intervertebral discs was examined by immunohistochemistry.
To determine the involvement of cathepsins in the pathomechanism of intervertebral disc degeneration by monitoring the immunolocalization of cathepsins in degenerated intervertebral disc tissue.
Cathepsins D, K, and L are enzymes that contribute to the matrix destruction seen in the articular cartilage affected by osteoarthritis and rheumatoid arthritis. However, little is known about the contribution of these cathepsins to intervertebral disc degeneration.
Paraffin-embedded sections of degenerated intervertebral disc tissue collected at the time of surgery (13 discs from 12 patients) were immunohistochemically stained with antibodies for cathepsins D, K, and L. For further characterization of the stained cells, immunohistochemical detection of CD68 and TRAP staining were performed.
Hematoxylin and eosin staining revealed obvious signs of degeneration in all sections. Cathepsins D and L were immunolocalized in disc fibrochondrocytes at various sites exhibiting degeneration. Cathepsins K were found in tartrate-resistant acid phosphatase-positive multinucleated cells, in particular near the cleft within the cartilaginous endplate. However, few cells were positive for these cathepsins in anulus fibrosus that maintained the lamellar structure of collagen fibers.
Marked expression of cathepsins D and L was observed at the site of degeneration. Cathepsins D and K localized in tartrate-resistant acid phosphatase-positive multinucleated cells existed at the cleft between the cartilaginous endplate and vertebral body. The site-specific localization of these cathepsins suggests the association of these proteinases with endplate separation and disorganization of the anulus fibrosus in degenerative spinal disorders.
采用免疫组织化学方法检测组织蛋白酶D、K和L在退变椎间盘内的定位。
通过监测组织蛋白酶在退变椎间盘组织中的免疫定位,确定组织蛋白酶在椎间盘退变发病机制中的作用。
组织蛋白酶D、K和L是导致骨关节炎和类风湿关节炎患者关节软骨基质破坏的酶。然而,关于这些组织蛋白酶在椎间盘退变中的作用知之甚少。
对手术时采集的退变椎间盘组织石蜡包埋切片(来自12例患者的13个椎间盘)进行免疫组织化学染色,使用组织蛋白酶D、K和L的抗体。为进一步鉴定染色细胞,进行CD68免疫组织化学检测和抗酒石酸酸性磷酸酶染色。
苏木精-伊红染色显示所有切片均有明显退变迹象。组织蛋白酶D和L免疫定位于退变的不同部位的椎间盘纤维软骨细胞中。组织蛋白酶K存在于抗酒石酸酸性磷酸酶阳性的多核细胞中,尤其是在软骨终板内的裂隙附近。然而,在维持胶原纤维层状结构的纤维环中,很少有细胞对这些组织蛋白酶呈阳性反应。
在退变部位观察到组织蛋白酶D和L的显著表达。存在于软骨终板和椎体之间裂隙处的抗酒石酸酸性磷酸酶阳性多核细胞中有组织蛋白酶D和K。这些组织蛋白酶的位点特异性定位表明这些蛋白酶与退行性脊柱疾病中终板分离和纤维环紊乱有关。