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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 To observe the expression of tryptase and chymase in the lung tissue of guinea pigs with anaphylactic shock and to provide an objective diagnosis basis for the death of anaphylactic shock. Methods Twenty-four healthy guinea pigs were randomly divided into experimental group and control group. Each group was subdivided into immediate death group, chilled 48h group and frozen 7d group, with 4 in each group. The experimental group 0.5mL human serum was diluted 1:10 with saline, injected into the skin of the guinea pigs after the sensitization 3 weeks mixed serum 1mL injected into the heart chamber induced anaphylactic shock and lethal; the control group instead of mixing saline serum. The blood and lung tissues of guinea pigs were extracted, and the expression of tryptase and chymase was observed by immunohistochemistry and image analysis. Results The number of tryptase and chymase positive cells in the lungs of guinea pigs in the control group were less and distributed around the small blood vessels and the trachea. In experimental group, the number of tryptase and chymase positive cells in the lung tissue increased obviously, the majority of cells were irregular in shape, and the positive staining particles escaped the mast cells and diffused to the interstitial space. Under the conditions of refrigerated 48h and frozen 7d, the expression of these two enzymes had no significant effect. Conclusion The expression of tryptase and chymase in the lung tissue of guinea pigs that were killed by anaphylactic shock is obviously enhanced. Under the condition of refrigerated storage for 48h and frozen for 7d, it can be used as a diagnostic basis for the death of anaphylactic shock.