Department of Orthopaedics, Peking University Third Hospital, Beijing, China.
Spine (Phila Pa 1976). 2010 Sep 15;35(20):E1018-24. doi: 10.1097/BRS.0b013e3181e198ab.
The study was undertaken to compare the expression and activity of tissue transglutaminase (TG2) in human ligamentum flavum cells derived from ossification of the ligamentum flavum (OLF) and non-OLF patients.
To determine whether TG2 is involved in the pathologic process of OLF.
OLF is a disease characterized by heterotopic formation of new bone in the flavum ligament. Recently, TG2 is proved to directly promote skeletal matrix mineralization and play an important role in the ossification. TG2 activity is vital to the differentiation of osteoblasts and the formation of mineralization. But whether TG2 is involved in the pathologic process of OLF is unknown. We investigated the relations between TG2 expression and OLF.
OLF and non-OLF cells were cultured and osteocalcin, bone morphogenetic protein-2(BMP-2) and TG2 mRNA expressions were assayed by reverse transcription polymerase chain reaction. Meanwhile, alkaline phosphatase activity and calcified nodules were compared between OLF and non-OLF cells. To detect TG2 expression, Western blot and immunohistochemical analysis were carried out, and TG2 activity was compared between OLF and non-OLF cells.
Our experiments demonstrated that OLF cells showed osteoblast-like activity and increased mRNA expression of BMP-2. More interesting, compared with non-OLF cells, OLF cells showed elevated expression levels of TG2 mRNA and protein, as well as enzyme activity.
TG2 expression and enzyme activity are upregulated in the OLF cells and TG2 may be involved in the pathologic process of OLF.
本研究旨在比较来源于黄韧带骨化(OLF)和非 OLF 患者的人黄韧带细胞中组织转谷氨酰胺酶(TG2)的表达和活性。
确定 TG2 是否参与 OLF 的病理过程。
OLF 是一种以黄韧带中异位形成新骨为特征的疾病。最近,研究证明 TG2 可直接促进骨骼基质矿化,并在骨化中发挥重要作用。TG2 活性对成骨细胞的分化和矿化的形成至关重要。但是,TG2 是否参与 OLF 的病理过程尚不清楚。我们研究了 TG2 表达与 OLF 之间的关系。
培养 OLF 和非 OLF 细胞,通过逆转录聚合酶链反应检测骨钙素、骨形态发生蛋白-2(BMP-2)和 TG2 mRNA 的表达。同时,比较 OLF 和非 OLF 细胞中的碱性磷酸酶活性和钙化结节。通过 Western blot 和免疫组织化学分析检测 TG2 表达,并比较 OLF 和非 OLF 细胞中的 TG2 活性。
我们的实验表明,OLF 细胞表现出成骨细胞样活性,并增加了 BMP-2 的 mRNA 表达。更有趣的是,与非 OLF 细胞相比,OLF 细胞中 TG2 mRNA 和蛋白的表达水平以及酶活性均升高。
OLF 细胞中 TG2 的表达和酶活性上调,TG2 可能参与 OLF 的病理过程。