TNF-α/TNFR1在内耳免疫反应中的表达  

TNF-α/TNFR1 Expression during the Immune Response of Inner Ear

作　　者：许丽娟[1] 龚树生[2] 汪吉宝[3] 黄翔[3] 陈佳[1] 陈小林[1] 

机构地区：[1]武汉科技大学附属天佑医院耳鼻咽喉科,武汉430064 [2]首都医科大学附属北京同仁医院耳鼻咽喉头颈外科,北京100730 [3]华中科技大学同济医学院附属协和医院耳鼻咽喉科,武汉430022

出　　处：《华中科技大学学报（医学版）》2010年第5期679-683,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：湖北省教育厅资助项目(No.Q20101109)

摘　　要：目的探讨内耳免疫反应过程TNF-α/TNFR1的表达及作用。方法选用雌性白色豚鼠45只,随机分为实验组29只,对照组16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),分别在内耳免疫术后1、3、5、7和14 d处死动物,取内耳免疫侧耳蜗做石蜡切片。通过免疫组化检测内耳TNF-α的表达,原位杂交检测内耳TNFR1mRNA的表达。结果对照组TNF-α蛋白表达阴性。实验组除内耳免疫第1天Corti器、侧壁和螺旋神经节与内耳免疫第3天侧壁外,其余切片都有TNF-α蛋白阳性表达,免疫组织化学染色平均吸光度值均显著高于对照组(均P<0.01)。对照组TNFR1 mRNA表达阴性。实验组除内耳免疫第1、3、5天Corti器外,其余切片均有TNFR1 mRNA阳性表达,TNFR1 mRNA原位杂交染色平均吸光度值均显著高于对照组(均P<0.01)。结论 TNF-α/TNFR1是调节内耳免疫反应的重要细胞因子和信号转导途径之一。Objective To investigate the expression and function of TNF-α/TNFR1 in the immune response of inner ear.Methods Forty-five healthy,female white guinea pigs were employed in this study.After being sensitized systematically with keyhole limpet hemocyanin（KLH）,the KLH-immunized animals were inoculated with the same antigen,and the control animals（n=16）were injected with PBS through cochlea basal turn.All animals were sacrificed respectively at the 1st,3rd,5th,7th,and 14th day after inner ear vaccination.Paraffin-embedded sections of cochleas from animals were prepared.Immunohistochemistry method was used to detect the expression of TNF-α,and in situ hybridization histochemistry method was used to detect the expression of TNFR1 mRNA in the inner ears.Results The TNF-α was expressed in the cochlea among the KLH immunized inner ears except for the cochlea at the 1st day after inner ear vaccination and the cochlear lateral wall at the 3rd day after inner ear vaccination.The cochlea in control group showed no apparent TNF-α immunoreactivity.Except for the cochlea at the 1st day after inner ear vaccination and the cochlear lateral wall at the 3rd day after inner ear vaccination,the incidence of TNF-α in the KLH immunized animals was significantly greater than in the control animals（P〈0.01）.The TNFR1 mRNA was expressed in the cochlea among the KLH immunized inner ears except for the Corti＇s organ at the 1st,3rd and 5th day after inner ear vaccination.No cells positive for TNFR1 mRNA were seen in the control inner ears.Except for the Corti＇s organ at the 1st,3rd,and 5th day after inner ear vaccination,the incidence of TNFR1 mRNA in the KLH immunized animals was significantly greater than in the control animals（P〈0.01）.Conclusion These findings suggest that the immune response of inner ear can induce apoptosis and TNF-α/TNFR1 pathway is one of important signal transportations during this course.

关 键 词：自身免疫 细胞凋亡 肿瘤坏死因子-Α 肿瘤坏死因子受体1 内耳 

分 类 号：R322.9[医药卫生—人体解剖和组织胚胎学]