豚鼠过敏性休克肺组织中类胰蛋白酶和胃促胰酶的表达

An immunohistochemical study of tryptase and chymase in the lungs of guinea pigs died of anaphylactic shock

目的观察豚鼠过敏性休克死亡肺组织中类胰蛋白酶和胃促胰酶的表达情况,试图为过敏性休克死亡提供客观的诊断依据。方法健康豚鼠24只,随机均分为实验组和对照组,每组再分死亡即时组、冷藏48h组和冷冻7d组,每组4只。实验组将0.5mL人混合血清用生理盐水1∶10稀释,注射于豚鼠后掌皮内,致敏后3周以人混合血清1mL注入心腔诱发过敏性休克致死;对照组采用生理盐水代替混合血清。提取豚鼠心血及肺组织,应用免疫组化染色和图像分析技术观察类胰蛋白酶和胃促胰酶的表达情况。结果对照组豚鼠肺组织中类胰蛋白酶和胃促胰酶阳性细胞数量较少,分布在小血管和小气管周围。实验组肺组织中类胰蛋白酶和胃促胰酶阳性细胞明显增多,多数细胞形态不规则,阳性染色颗粒脱出肥大细胞并弥散到组织间隙。冷藏48h和冷冻7d的条件下对这两种酶的表达无明显影响。结论过敏性休克致死豚鼠肺组织中类胰蛋白酶和胃促胰酶的表达明显增强,在冷藏48h和冷冻7d内的条件下,可作为过敏性休克死亡的一项诊断依据。

Objective The aim of this study is attempted to provide an objective evidence for diagnosis of anaphylactic shock.Methods 24 healthy guinea pigs were randomly divided into experimental group and control group.For experimental group,animals were sub-grouped into animals of instantaneous death from anaphylactic shock,animals kept at 4℃ for 48h after death and those kept at-20℃ for 7d after death.For the control animals sub-groups were the same as the experimental animals.4 animals was in each sub-group.Anaphylacatic shock was induced in the experimental rabbits by intraventricular injection of human sera.Tryptase and chymase in the lungs were detected by immunohistochemical staining.Image analysis was performed.Results Tryptase-or chymase-positive mast cells were detected in the lungs in control animals,which were scattered around small vessels and bronchioles.Tryptase-positive or chymase-positive cells were significantly increased in number,and tryptase-or chymase-stained granules free from mast cells were detetced in the interstitial space in the lungs in the animals of deaths from anaphylactic shock.The preserving conditions did not affect the immunostaining for tryptase or chymase under 4℃ for 48h or-20℃ for 7d.Conclusion Immunostaining for tryptase and chymase may provide an objective method for diagnosis of anaphylactic shock.