Department of Oral and Maxillofacial Surgery, Wakayama Medical University, Japan.
Eur J Histochem. 2010 Dec 15;54(4):e50. doi: 10.4081/ejh.2010.e50.
The present study aimed at investigating the expression of a hyaluronan synthase (HAS) 3 in tissue samples of deformed human temporomandibular joint (TMJ) discs and cells obtained from the discs. Fifteen adult human TMJ discs (twelve diseased discs and three normal discs) were used in this study. The twelve diseased discs were obtained from twelve patients with internal derangement (ID) of TMJ. These patients all had anteriorly displaced discs and deformed discs. The tissues were immunohistochemically stained using HAS3 antibodies. In addition, the subcultured TMJ disc cells under both normal and hypoxic conditions (O2: 2%) were incubated for 3, 6, 12, and 24 h after addition of interleukin-1β (IL-1β) (1 ng/mL). Subsequently, the expression of HAS3 was examined using real-time reverse transcription-polymerase chain reaction (RT-PCR). The control group showed from negative to weak positive reactions for HAS3 on immunohistochemical staining. The discs extracted from twelve cases with ID presented from moderate to strong positive reactions for HAS3. The quantity of HAS3 mRNA was compared with a control group, and showed a 204-fold increase at 3 h, a 26-fold increase at 6 h, a 2.5-fold increase at 12 h and a 32-fold increase at 24 h under hypoxia with the addition of IL-1β. The expression of HAS3 mRNA was significantly enhanced at 3 h and 24 h. The results obtained suggest that HAS3 is related to the pathological changes of human TMJ discs affected by ID.
本研究旨在探讨透明质酸合酶 (HAS) 3 在变形人类颞下颌关节 (TMJ) 盘组织样本和从盘获得的细胞中的表达。本研究使用了 15 个人类 TMJ 盘(12 个患病盘和 3 个正常盘)。这 12 个患病盘来自 12 个 TMJ 内部紊乱的患者。这些患者均有前移位盘和变形盘。使用 HAS3 抗体对组织进行免疫组织化学染色。此外,在添加白细胞介素-1β (IL-1β) (1 ng/mL) 后,将正常和缺氧条件 (O2: 2%) 下培养的 TMJ 盘细胞亚培养物分别孵育 3、6、12 和 24 h,然后使用实时逆转录聚合酶链反应 (RT-PCR) 检查 HAS3 的表达。对照组在免疫组织化学染色中显示 HAS3 呈阴性至弱阳性反应。从 12 例 ID 患者提取的盘显示 HAS3 呈中度至强阳性反应。与对照组相比,在添加 IL-1β 后缺氧条件下 3 h 时 HAS3 mRNA 增加了 204 倍,6 h 时增加了 26 倍,12 h 时增加了 2.5 倍,24 h 时增加了 32 倍。HAS3 mRNA 的表达在 3 h 和 24 h 显著增强。研究结果表明,HAS3 与 ID 影响的人类 TMJ 盘的病理变化有关。