单染与复染在观察SIRT1基因敲除小鼠骨关节炎切片中的差异比较

Observation of the articular cartilage of osteoarthritis in SIRT1 gene knock-out mice through single staining and compound staining

目的通过SIRT1基因敲除建立相关小鼠膝关节骨关节炎模型,在此基础上观察各种不同单独染色或复合染色技术在观察软骨组织切片形态学中的差异。方法将小鼠膝关节组织标本分为2组:SIRT1-/-小鼠假手术组(n=6,A组),SIRT1-/-小鼠骨关节炎模型组(n=6,B组)。通过前交叉韧带横断加内侧半月板切除术建立膝关节骨关节炎模型,行HE染色、番红O-固绿染色、番红O-阿利新蓝染色、番红O染色、固绿染色、阿利新蓝染色,6种单独染色或复合染色方式观察膝关节软骨组织形态学变化。结果番红O-固绿染色、番红O-阿利新蓝染色在观察软骨细胞的细胞形态、软骨分层结构的情况、II型胶原纤维的显示、潮线及软骨下骨的改变中,效果更好;而番红O染色、阿利新蓝染色在观察软骨组织缺失方面有一定的优势。结论在观察SIRT1基因敲除小鼠膝关节骨关节炎软骨组织切片中,与单独染色相比,复合染色在获得软骨结构的各种信息方面优势更加明显。

Objective To establish osteoarthritis model of the knee joint in mice on the basis of knocking out SIRT1 gene and to observe the differences in the morphology of the cartilage tissue using single staining and compound staining. Methods The knee joint specimens were divided into two groups： SIRT1-/-control group（ group A,n = 6）and SIRT1-/-osteoarthritis model group（ group B,n = 6）. The knee anterior cruciate ligament was traversed,and the ipsilateral medial meniscus was cut to establish an osteoarthritis model of knee joint. HE staining,safranin O-fast green staining,safranin O-alcian blue staining,safranin O staining,fast green staining,alcian blue staining were used to observe the morphological changes in the articular cartilage of the knee. Results Safranin O-fast green staining and safranin Oalcian blue staining showed better results in observation of the morphology of chondrocytes,the structure of cartilage layers,the presence of type II collagen,tide line and the changes of subchondral bone. While the safranin O staining and alcian blue staining had certain advantages in the observation of the defects of cartilage tissue. Conclusions Compared with thesingle staining,the compound staining used in this study have obvious advantages in obtaining useful information of the cartilage structure in the observation of morphology of cartilage tissues in SIRT1 gene knock-out mice.