绵羊痒螨重组精氨酸激酶诱导兔皮肤嗜酸性粒细胞的聚集  

Recombinant Arginine Kinase of Psoroptes ovis Induces Cutaneous Eosinophil Recruitment in Rabbit

作　　者：黄翠锐 郝桂英[2] 周宇 田焱[1] 杨富升 唐丽[1] 杨光友[1] 谢跃[1] 何冉 徐璟[1] 古小彬[1] HUANG Cuirui;HAO Guiying;ZHOU Yu;TIAN Yan;YANG Fusheng;TANG Li;YANG Guangyou;XIE Yue;HE Ran;XU Jing;GU Xiaobin(College of Veterinary Medicine,Sichuan Agricultural University,Chengdu 611130,China;School of Animal Science of Xichang University,Xichang 615013,China)

机构地区：[1]四川农业大学动物医学院,成都611130 [2]西昌学院动物科学学院,西昌615013

出　　处：《畜牧兽医学报》2023年第5期2186-2194,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：四川省科技计划重点研发项目(2019YFN0155);四川农业大学学科双支计划(2221993268)。

摘　　要：本研究旨在探究绵羊痒螨精氨酸激酶(arginine kinase of Psoroptes ovi s,Pso AK)对兔皮肤嗜酸性粒细胞聚集的影响。以永生化角质形成细胞(HaCaT)和痒螨致敏兔分别为体外模型和体内模型,重组Pso AK(r Pso AK)分别刺激HaCaT细胞和皮内注射痒螨致敏兔后,采用RT-qPCR检测重组蛋白处理后HaCaT细胞和注射部位兔皮肤组织中与嗜酸性粒细胞聚集相关因子(IL-4、IL-5、IL-13、TSLP、IL-25、IL-33、CCL5、CCL11、IL-4R)转录水平,以及通过变色酸2R特殊染色观察,并统计兔皮肤组织中聚集的嗜酸性粒细胞数量。结果表明,与空白和载体蛋白对照组相比,0.1μg·mL^(-1)r Pso AK可显著促进HaCaT细胞中IL-13、IL-33、CCL5和CCL11转录水平的增加(P<0.05),以及兔皮肤内IL-4R和CCL5转录水平的增加(P<0.01),其他因子无显著变化(P>0.05);同时,与PBS和载体蛋白对照组相比,r Pso AK可诱导皮肤中嗜酸性粒细胞的数量极显著增加(P<0.001)。综上所述,Pso AK能够促进部分嗜酸性粒细胞聚集相关细胞因子和趋化因子转录水平的增加,且诱导兔皮肤内嗜酸性粒细胞的聚集,表明Pso AK是引发痒螨病皮损区嗜酸性粒细胞聚集的抗原分子,其参与了绵羊痒螨感染的致病过程。The object of this study was to investigate the effect of arginine kinase of Psoroptes ovi s(Pso AK)on cutaneous eosinophil recruitment in rabbit.I n vitro of HaCaT cells and in vivo models of P.ovis-sensitization rabbits,qRT-PCR analysis were used to evaluate whether the recombinant Pso AK(r Pso AK)has effect on the eosinophils-related genes expression in vitro and in vivo,such as IL-4,IL-5,IL-13,TSLP,IL-25,IL-33,CCL5,CCL11,IL-4R.Additionally,chromotrope-2R staining was employed to evaluate whether r Pso AK can induce eosinophil recruitment to skin in vivo.Compared to the blank and pET32a control groups,0.1μg·mL^(-1) r Pso AK significantly promoted the relative transcript levels of IL-13,IL-33,CCL5 and CCL11 mRNA in HaCaT cells(P<0.05).In vivo,r Pso AK was also significantly upregulated in the mRNA levels of IL-4R and CCL5 in rabbit skin compared with the blank and pET32a control groups(P<0.01),and there was no significant effect on other factors(P>0.05).Meanwhile,r Pso AK significantly induced the increased number of eosinophil recruitment in rabbit skin as compared with PBS and vector control groups(P<0.001).In conclusion,Pso AK can promote the increase of some eosinophil-related genes expressions and induce cutaneous eosinophil recruitment in rabbit,and serves as a molecule contributing to skin eosinophils accumulation in P.ovis infection and involves in the pathogenesis of P.ovis infestation.

关 键 词：绵羊痒螨 重组精氨酸激酶 兔 皮肤 嗜酸性粒细胞聚集 

分 类 号：S852.746[农业科学—基础兽医学]