重组腺病毒介导SOX-9基因对颈椎关节突软骨细胞凋亡的影响

目的 探讨重组腺病毒介导的人SOX-9(AdSOX-9)对人的颈椎关节突关节软骨细胞凋亡细胞数目及Caspase-3基因表达的作用及相关意义,更好理解颈椎软骨退变的发生机制.方法 应用AdSOX-9按照不同浓度及作用时间处理体外培养软骨细胞,应用TdT介导dUTP-生物缺口末端标记方法(TUNEL)及免疫染色方法观察凋亡细胞数量及Caspase-3基因表达变化情况.结果 25MOI、50MOI剂量作用组TUNEL阳性细胞较空白对照组均明显增加;AdSOX-9转染软骨细胞预处理后,空白对照组、25MOI作用组、50MOI三种处理组Caspase-3表达均明显增加,各时点较空白对照组比较,存在显著性差异(P＜0.05).结论 SOX-9基因对软骨凋亡细胞数目、Caspase-3凋亡基因的影响,存在时间-剂量依赖关系,为软骨细胞退变发病机制研究提出新的思考.

The regulating effects of adSOX-9 on apoptosis in the hyaline cartilage cells of human cervical zygapophysial joints

Objective To investigate the role of SOX-9(SRY-related high mobility group box gene 9)regulating the apoptosis in the hyaline cartilage cells of human cervical zygapophysial joints. Methods In the experiment,human hyaline chondrocytes were harvested enzymatically and cultured in DMEM supplemented with 20% fetus calf serum,varying concentration of SOX-9 was added to the medium. The amount of apoptosis cell were assessed by TUNEL and immunostaining. Results SOX-9 with different dosages and action times can increase the amount of apoptosis cell and the expression level of Caspase-3. Their difference was significant(P<0.05)and the level of Caspase-3 expression have significant coherence with the dose of SOX-9. Conclusion Our results show that normal human chondrocytes express small amount of apoptosis cell,SOX-9 can cause a dose-dependent stimulation on Caspase-3 expression in human chondrocyte,SOX-9 may play a crucial role in the human cervical degenerative disease.