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目的 研究内耳免疫反应过程中是否存在细胞凋亡 ,以及细胞凋亡是否与Caspase -3信号转导有关。方法 选用雌性白色豚鼠 16只 ,随机分为实验组和对照组各 8只 ,以钥孔虫戚血蓝蛋白 (keyholelimpethemocyanin ,KLH)全身免疫后 ,实验组以相同抗原进行内耳免疫 ,对照组内耳注射等量的磷酸盐缓冲生理盐水 (PBS) ,在内耳免疫 5d后处死动物 ,取内耳免疫侧耳蜗做石蜡切片。通过TUNEL法检测内耳凋亡细胞 ,免疫组化检测内耳Caspase -3的表达。结果 实验组Corti器 ,血管纹、螺旋韧带和螺旋神经节存在TUNEL染色阳性细胞 ,而对照组仅在支持细胞、血管纹和螺旋神经节中发现极少数TUNEL染色阳性细胞。免疫组化染色实验组Caspase -3表达阳性 ,而对照组表达阴性。结论 内耳免疫反应可以诱导细胞凋亡的发生 ;内耳免疫反应诱导细胞凋亡的发生中有Caspase -3的激活。
Objective To investigate the existence of apoptosis in the inner ear immune response and whether apoptosis is related to Caspase -3 signal transduction. Methods Twenty-six female white guinea pigs were randomly divided into experimental and control groups. After immunization with keyhole limpet hemocyanin (KLH), the experimental group received the same inner ear immunization and the control group received intra-ear injection An equal volume of phosphate buffered saline (PBS) was used to kill the animals 5 days after immunization in the inner ear. The inner ear immunocephalic cochlea were paraffin-embedded. Apoptotic cells were detected by TUNEL method. The expression of Caspase-3 in the inner ear was detected by immunohistochemistry. Results TUNEL-positive cells were present in the Corti vessels, vascular lines, spiral ligaments and spiral ganglia in the experimental group, but only a few TUNEL-positive cells in the control group were found in the supporting cells, vasculature and spiral ganglion. Immunohistochemical staining Caspase -3 expression in the experimental group, while the control group negative expression. Conclusion The immune responses in the inner ear can induce the apoptosis of cells. The activation of Caspase-3 in the inner ear induced by immune response in the inner ear can induce the apoptosis.