酸敏感离子通道1a在大鼠椎体终板软骨细胞的表达及其意义

Expression of acid-sensing ion channel 1a in endplate chondrocytes of the vertebral body in rats

目的探讨大鼠椎体终板软骨细胞酸敏感离子通道1a(ASIC1a)的表达及其意义。方法选用成年SD大鼠10只。分离、培养椎体终板软骨细胞以及传代培养,倒置显微镜下观察细胞形态,甲苯胺蓝染色对终板软骨细胞进行鉴定。利用PT-PCR和Western blotting检测ASIC1a在大鼠椎体终板软骨细胞mRNA及其蛋白的表达,免疫荧光双标观察ASIC1a在终板软骨细胞的细胞定位。结果形态学及甲苯胺蓝染色阳性显示,培养的细胞为软骨细胞,PT-PCR和Western blotting显示,大鼠椎体终板软骨细胞存在ASIC1a mRNA及其蛋白的表达,免疫荧光显示,ASIC1a在终板软骨细胞膜有阳性表达。结论 ASIC1a在大鼠椎体终板软骨细胞的表达,对认识ASIC1a功能及寻找新抗椎间盘退变靶点有重要意义。

Objective To explore the expression of acid-sensing ion channels la （ASICla） in endplate chondroeytes of the vertebral body in rat. Methods Endplate chondroeytes were isolated from the 10 adult SD rats, and their phenotypes were determined by an inverted microscope and toluidine blue staining. RT-PCR and Western blotting were used to detect ASIC1 a in endplate chondrocytes of the rat vertebral body. Fluorescence immunoeytoehemistry double marker was used to the location of ASIC1 a in the cells. Results Positive staining for glycosaminoglyean was seen in endplate chondrocytes. RT-PCR analysis showed that ASICla mRNA were present in endplate chondrocytes of the vertebral body. The total RNA, and the corresponding protein were detected. Conclusion The ASICIa expresses in endplate chondrocytes of the vertebral body, which may be regarded as a new target of intervertebral disc degeneration （IVDD）.