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目的 了解抗原免疫对热休克蛋白 70 (heatshockprotein 70 ,HSP70 )在豚鼠耳蜗中表达的影响。方法 以同种异体内耳组织为抗原免疫豚鼠 ,用免疫组织化学方法检测豚鼠耳蜗中HSP70的表达并进行计算机图像处理 ;对豚鼠膜迷路组织进行免疫转印分析以测定HSP71 ;以纯化的人类重组HSP71为抗原 ,用免疫转印技术 (Westernblot)检测动物血浆中HSP71抗体。结果 HSP70在正常豚鼠耳蜗中呈低水平表达。抗原免疫后 ,实验组动物耳蜗HSP70免疫活性较对照组明显增强 ,两组间的吸光度值差异有显著性 (P <0 0 1 ) ;免疫后大多数动物 (1 2 / 2 0 )体内产生了抗HSP71抗体 ,实验组HSP71抗体阳性率显著高于对照组 (P <0 0 1 )。结论 同种膜迷路组织作为抗原免疫豚鼠能够诱导HSP70在耳蜗内合成增加 ;表明HSP70与自身免疫性内耳病有一定联系。
Objective To investigate the effect of antigen immunization on the expression of heat shock protein 70 (HSP70) in cochlea of guinea pigs. Methods The guinea pigs were immunized with allogenic inner ear tissue antigen and the expression of HSP70 in the cochlea of guinea pigs was detected by immunohistochemistry. The expression of HSP70 in guinea pig cochlea was detected by immunoblotting analysis. The purified recombinant human HSP71 As antigen, HSP71 antibody in plasma of animals was detected by immunoblotting (Western blot). Results HSP70 expression in the normal guinea pig cochlea was low. After antigen immunization, the immunocompetence of HSP70 in experimental group was significantly higher than that in control group, the difference was significant (P <0.01); after immunization, most of the animals (12/200) produced in vivo The anti-HSP71 antibody, the positive rate of HSP71 antibody in the experimental group was significantly higher than that of the control group (P <0.01). Conclusion The same kind of membrane labyrinth tissue as antigen immunized guinea pigs can induce the synthesis of HSP70 in the cochlea, which indicates that there is a certain relationship between HSP70 and autoimmune inner ear disease.