Ⅰ型胶原在人颞下颌关节盘穿孔病变细胞中的表达

The changes of type Ⅰ collagen expression in fibroblast-like cells from temporomandibular joint disc perforation

目的:研究颞下颌关节盘穿孔时关节盘细胞合成及分泌Ⅰ型胶原能力的改变及其在颞下颌关节盘穿孔发生中的作用。方法:收集颞下颌关节髁突肥大(对照组)及颞下颌关节盘穿孔(实验组)患者因手术切除的手术标本,采用酶消化法原代培养,培养的细胞经甲苯胺蓝染色及免疫荧光染色鉴定为成纤维样细胞。应用酶联免疫吸附测定法检测关节盘细胞培养上清液中Ⅰ型胶原蛋白的含量。采用蛋白质印迹法分析细胞内Ⅰ型胶原蛋白的表达。结果:实验组及对照组的颞下颌关节盘均可培养出成纤维样细胞,甲苯胺蓝染色、Ⅰ型胶原及Ⅱ型胶原免疫荧光染色均表达阳性。对照组细胞培养48 h后上清液中Ⅰ型胶原的含量显著高于实验组(P<0.05)。蛋白质印迹法检测结果显示,实验组细胞内Ⅰ型胶原的蛋白表达明显低于对照组(P<0.05)。结论:颞下颌关节盘细胞表达及分泌Ⅰ型胶原含量减少,Ⅰ型胶原表达降低可能导致人颞下颌关节盘穿孔的发生及进展。

Objective： To study the changes of type Ⅰ collagen expression in fibroblast-like cells from temporomandibular joint disc perforation and investigate the effect of type Ⅰ collagen in the occurrence of temporomandibular joint disc perforation. Methods： The fibroblastic-like cells in experimental group from the surgical removed temporomandibular joint disc perforation tissue were cultured,and the cells from temporomandibular joint condyle hyperplasia were also cultured as the control group. The cultured cells were identified as fibroblastic-like cells by toluidine blue and immunofluorescence staining.The expression of intracellular type Ⅰ collagen was detected with Western Blotting and the content of extracellular type Ⅰcollagen was examined by enzyme linked immunosorbent assay. Results： Fibroblastic-like cells in two groups were positive by toluidine blue,and the collagen-Ⅰ and collagen-Ⅱ were positive by immunofluorescence staining. The content of extracellular collagen-Ⅰ was significantly higher than that in the control group（ P〈0. 05）. The expression of typeⅠcollagen in cells from joints with disc perforation was lower than that from normal joints（ P〈0. 05）. Conclusion： The expression level of collagen-Ⅰdecreases significantly in fibroblastic-like cells from temporomandibular joint disc perforation tissue,which may lead to the development and progression of temporomandibular joint disc perforation.