实验性急性中耳细菌感染诱发的热休克反应

Heat shock response in experimental acute middle ear bacterial infection

目的探讨细菌抗原和内耳抗原之间的联系及热休克反应对其抗原的影响。方法用抗哺乳类热休克蛋白（ｈｅａｔｓｈｏｃｋｐｒｏｔｅｉｎ，ｈｓｐ）７０抗体识别绿脓杆菌、变形杆菌、大肠杆菌、金黄色葡萄球菌和肺炎克雷白杆菌抗原，进行免疫转印分析，测试正常豚鼠膜迷路中ｈｓｐ７０的表达水平。用肺炎克雷白杆菌制作急性中耳感染动物模型，观察细菌和内耳与哺乳类ｈｓｐ７０相关的蛋白表达水平动态变化。结果抗哺乳类ｈｓｐ７０单克隆抗体识别上述各种细菌抗原均非ｈｓｐ７０家族，正常豚鼠膜迷路表达ｈｓｐ７０水平极低。肺炎克雷白杆菌侵入中耳后，其自身出现热休克反应，表达ｈｓｐ水平增高，用抗ｈｓｐ７０单抗识别的表位有分子量７００００和１７０００的蛋白分子。分子量７００００蛋白表达水平为注射后３ｄ最高，１７０００蛋白表达水平于注射后５ｄ最高。豚鼠中耳受肺炎克雷白杆菌感染后，急性期内耳主要表达ｈｓｐ７０蛋白和与之有相关表位的ｈｓｐ３１，７００００蛋白的表达水平明显低于３１０００。结论细菌侵入机体后其自身发生热休克反应，机体内耳ｈｓｐ表达水平也相对增高，这些存在共同表位的ｈｓｐ的高表达为微生物感染诱发内耳自身免疫反应提供了物质基础。小分子ｈｓｐ有可能参与内?

Objective To investigate the association between bacteria antigen and inner ear antigen and effects of stress on the expression spectrum and level of the antigen bearing hsp70 related epitope. Methods Use monoclonal antibody against mammiferous hsp70 to recognize the related antigens of Bacillus pyocyaneus,Bacillus proteus,Bacillus coli,Staphylococcus aureus and Klebsiella pneumoniae with modified western blotting. The expression level of hsp70 in unstressed guinea pig inner ear was tested.An animal model of middle ear acute infection by Klebsiella pneumoniae was established. The dynamic changes of mammiferous hsp70 epitope related proteins were tested with modified western blotting. Results All the bacteria antigens recognized by anti mammiferous hsp70 antibody were not of the hsp70 family and the expression level of hsp70 in normal guinea pig was extraordinarily low.Klebsiella pneumoniae expressed 70 000 dalton and 17 000 dalton hsp after its invasion into guinea pig middle ear. 70 000 dalton antigen expressed most intensely at 3rd day of invasion and then fell.17 000 dalton antigen was expressed most intensely at 5th day of invasion and then fell. The expression of 70 000 dalton hsp was much weaker than 31 000 dalton hsp at acute periods of middle ear infection. Conclusions The elevated expression of hsp both in bacteria and inner ear during middle ear acute infection provides material base for immune impairment on inner ear induced by bacterial infection.The low molecular weight hsp may be involved in the inner ear autoimmune impairment.