半胱氨酸天冬氨酸蛋白酶3在亚洲带绦虫实验感染乳猪肝组织的表达

发布时间：2018-05-21 18:16

本文选题：亚洲带绦虫 + 半胱氨酸天冬氨酸蛋白酶　；参考：《中国人兽共患病学报》2017年04期

【摘要】：目的分析亚洲带绦虫实验感染乳猪后囊尾蚴寄生处肝脏组织半胱氨酸天冬氨酸蛋白酶3(Cysteine aspartyl proteinase 3,Caspase-3)的表达变化情况,为探讨Caspase-3在亚洲带绦虫感染致乳猪肝损伤中的作用提供基础资料。方法将采自贵州省都匀市良亩乡的亚洲带绦虫孕节解剖后,以生理盐水反复清洗并离心收集虫卵。20d龄约克施格杂交乳猪12头随机分为实验组和对照组各6头,实验组以定量虫卵15万个/头灌胃感染,于感染后第15d和第75d解剖乳猪,取实验组囊尾蚴寄生处肝组织和对照组对应部位肝组织,运用实时荧光定量PCR和免疫印迹方法分别定性定量分析Caspase-3在实验组和对照组中mRNA水平、蛋白水平的表达,并进一步运用免疫组织化学方法对Caspase-3在实验组和对照组中的蛋白表达水平和蛋白表达部位进行分析和定位。结果感染后第15d,实时荧光定量PCR结果显示实验组Caspase-3mRNA水平低于对照组(P=0.011),免疫印迹结果和免疫组化结果均显示实验组Caspase-3蛋白表达量低于对照组(P=0.008,P=0.004);此外,实验组感染后第75d的Caspase-3mRNA水平、蛋白表达量均高于实验组感染后第15d的样本(P=0.018,P=0.003,P=0.002);免疫组织化学结果显示Caspase-3阳性染色主要定位在肝细胞胞质中,呈黄色或棕黄色。感染后第75d实验组和对照组各项结果差异均无统计学意义。结论 Caspase-3在亚洲带绦虫感染早期的乳猪肝组织中低表达,而感染晚期正常表达,提示Caspase-3可能参与了亚洲带绦虫致乳猪早期肝损伤的调节。
[Abstract]:Objective to analyze the expression of cysteine aspartate protease 3(Cysteine aspartyl proteinase 3 (Caspase-3) in the liver of cysticercaria after Taenia asiatica infection. To investigate the role of Caspase-3 in liver injury induced by Taenia asiatica infection in suckling pigs. Methods after dissecting the pregnant nodes of Taenia asiatica collected from Liangmu Township, Duyun City, Guizhou Province, 12 Yorkshire crossbred suckling pigs of 20 days old were randomly divided into two groups: experimental group and control group. The experimental group was infected with 150000 eggs per head by stomach, and then dissected the suckling pigs on the 15th and 75th day after infection. The liver tissues of the experimental group and the control group were taken from the parasitic part of the cysticercaria and the corresponding liver tissue of the control group. The expression of mRNA and protein in experimental group and control group were analyzed qualitatively and quantitatively by real-time fluorescence quantitative PCR and Western blot, respectively. The protein expression level and protein expression site of Caspase-3 in experimental group and control group were analyzed and located by immunohistochemical method. Results on the 15th day after infection, the results of real-time fluorescence quantitative PCR showed that the level of Caspase-3mRNA in the experimental group was lower than that in the control group (0.011%), the expression of Caspase-3 protein in the experimental group was lower than that in the control group (P 0.008) and the level of Caspase-3mRNA in the experimental group was lower than that in the control group on the 75th day after infection, the expression of Caspase-3 protein in the experimental group was lower than that in the control group. The protein expression was higher than that in the experimental group on the 15th day after infection. The immunohistochemical results showed that the Caspase-3 positive staining was mainly located in the cytoplasm of hepatocytes, yellow or brown. There was no significant difference between the experimental group and the control group on the 75 th day after infection. Conclusion the expression of Caspase-3 is low in the early stage of Taenia asiatica infection, but normal in the late stage of infection, suggesting that Caspase-3 may be involved in the regulation of early liver injury induced by Taenia asiatica.
【作者单位】：贵州医科大学寄生虫学教研室;贵州医科大学病原生物学实验室;
【基金】：国家自然科学基金项目(No.81160205) 贵州省优秀科技教育人才省长资金项目(黔省专合字[2008]55号) 贵州省留学人员科技创新项目(黔人项目资助合同[2015]1号)联合资助~~
【分类号】：R383.3

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