摘要
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Fas和FasL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(keyhole limpet hemocyanin,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(phosphate buffered saline,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。通过脱氧核糖核苷酸末端转移酶介导的缺口末端标记技术(terminal-deoxynucleotidyl transferase mediated nick end labeling,TUNEL)检测内耳凋亡细胞,免疫组化检测内耳Fas和FasL的表达。结果实验组豚鼠内耳Corti器毛细胞,血管纹的缘细胞和螺旋神经节细胞存在TUNEL染色阳性细胞,而对照组动物切片仅在支持细胞、血管纹和螺旋神经节细胞中发现极少数TUNEL染色阳性细胞。免疫组化染色实验组Corti器、螺旋神经节细胞、血管纹和螺旋韧带Fas和FasL蛋白表达阳性,而对照组只有螺旋神经节细胞和血管纹有较弱的Fas蛋白表达,FasL蛋白表达阴性。结论内耳免疫反应可诱导细胞凋亡的发生,Fas-FasL途径是参与此过程重要的信号转导途径之一。
Objective To investigate whether apoptosis.is invovled in the pathogenesis of the immune response of the inner ear, and to explore the relation between signal transportation of Fas, FasL and apoptosis. Methods Sixteen healthy, female guinea pigs were employed in the experiment. Sensitized systematically with keyhole limpet hemocyanin （KIH）, the KLH-immunized animals were inoculated with the same antigen,,-while the control animals were injected PBS through cochlea basal turn. The animals were sacrificed 5 days after inner ear vaccination. Paraffin sections of cochleas were stained by TUNEL assay to identify inner ear cells undergoing apoptosis, and immunohistochemistry was used to detect the expression of Fas and FasL in the inner ears. Results TUNEL-positive cells were found in the KLH-immunized inner ears but not in the control inner ears except a few positive cells in the supporting cells, the stria vascularis cells and the spiral ganglion cells. The positive cells were the hair cells in the Corti＇s organ, the marginal cells in the stria vascularis and the neurons in the spiral ganglion. High expression of Fas and FasL could be detected in the Corti＇s organ, the stria vascularis, the spiral ligament and the neurons of the spiral ganglion in the KLH-immunized inner ears. Low expression of Fas could be detected in the stria vascularis and the neurons of the spiral ganglion in the control inner ears, but no positive FasL. cells were found in the control inner ears. Conclusion These findings suggest that apopt.osis is involved in the pathogenesis of the immune response of the inner ear and the Fas-FasL is one of the important, pathways of signal transportation.
出处
《中国组织化学与细胞化学杂志》
CAS
CSCD
2005年第6期629-634,共6页
Chinese Journal of Histochemistry and Cytochemistry
