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一种新型 JNK 通过调节牡蛎 Crassostrea gigas 中的 IL 表达参与免疫反应。

A novel JNK is involved in immune response by regulating IL expression in oyster Crassostrea gigas.

机构信息

Liaoning Key Laboratory of Marine Animal Immunology, Dalian Ocean University, Dalian 116023, China; Liaoning Key Laboratory of Marine Animal Immunology & Disease Control, Dalian Ocean University, Dalian 116023, China.

Liaoning Key Laboratory of Marine Animal Immunology, Dalian Ocean University, Dalian 116023, China.

出版信息

Fish Shellfish Immunol. 2018 Aug;79:93-101. doi: 10.1016/j.fsi.2018.05.017. Epub 2018 May 8.

Abstract

The c-Jun N-terminal kinase (JNK) is a member of mitogen-activated protein kinases (MAPK) highly conserved from yeast to mammals and participates in regulating many physiological and pathological processes. In the present study, a novel JNK was identified from oyster Crassostrea gigas (designated as CgJNK) and its biological functions were investigated in response against lipopolysaccharide (LPS) stimulation. The CgJNK consists of 415 amino acids, which includes a serine/threonine protein kinase (S_TKc) domain with a conserved Thr-Pro-Tyr (TPY) motif. Phylogenetic analysis revealed that CgJNK shared high similarity with other members of the JNK subfamily. CgJNK mRNA was detected in all the tested tissues and CgJNK mRNA expression levels in hemocytes were significantly up-regulated from 6 to 72 h after LPS stimulation and reached the highest level (16.1-fold, p < 0.01) at 24 h. The phosphorylation level of CgJNK in C. gigas hemocytes was increased at 2 h after LPS stimulation. The subcellular localization of CgJNK phosphorylation in hemocytes was analyzed after LPS stimulation, and CgJNK phosphorylation could be detected in both cytoplasm and nucleus of oyster hemocytes at 2 h post LPS stimulation. Additionally, the interleukins (CgILs) were detected in hemocytes of CgJNK-knockdown oysters. CgIL17-1, CgIL17-2, CgIL17-4 and CgIL17-6 transcripts were decreased significantly in CgJNK-knockdown oysters at 24 h post LPS stimulation. In summary, these results suggested that CgJNK played an important role in the immune response of oysters by regulating IL expression.

摘要

c-Jun N-端激酶(JNK)是一种高度保守的丝裂原活化蛋白激酶(MAPK),从酵母到哺乳动物都有参与,并参与调节许多生理和病理过程。本研究从牡蛎(Crassostrea gigas)中鉴定出一种新型 JNK(命名为 CgJNK),并研究了其对脂多糖(LPS)刺激的反应中的生物学功能。CgJNK 由 415 个氨基酸组成,其中包括一个丝氨酸/苏氨酸蛋白激酶(S_TKc)结构域,具有保守的 Thr-Pro-Tyr(TPY)基序。系统发育分析表明,CgJNK 与 JNK 亚家族的其他成员具有高度相似性。CgJNK mRNA 在所有测试的组织中均有检测到,并且在 LPS 刺激后 6 至 72 小时,血细胞中的 CgJNK mRNA 表达水平显著上调,在 24 小时达到最高水平(16.1 倍,p<0.01)。在 LPS 刺激后 2 小时,牡蛎血细胞中的 CgJNK 磷酸化水平增加。在 LPS 刺激后分析了血细胞中 CgJNK 磷酸化的亚细胞定位,在 LPS 刺激后 2 小时,可以在牡蛎血细胞的细胞质和细胞核中检测到 CgJNK 磷酸化。此外,还检测了 CgJNK 敲低牡蛎血细胞中的白细胞介素(CgILs)。在 LPS 刺激后 24 小时,CgJNK 敲低牡蛎中的 CgIL17-1、CgIL17-2、CgIL17-4 和 CgIL17-6 转录本显著降低。综上所述,这些结果表明,CgJNK 通过调节 IL 表达在牡蛎的免疫反应中发挥重要作用。

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