Abstract：
BACKGROUND: Cystatin is a protease inhibitor that also regulates genes expression linked to inflammation and plays a role in defense and regulation.
RESULTS: Cystatin 10 (Smcys10) was cloned from Scophthalmus maximus and encodes a 145 amino acid polypeptide. The results of qRT-PCR showed that Smcys10 exhibited tissue-specific expression patterns, and its expression was significantly higher in the skin than in other tissues. The expression level of Smcys10 was significantly different in the skin, gill, head kidney, spleen and macrophages after Vibrio anguillarum infection, indicating that Smcys10 may play an important role in resistance to V. anguillarum infection. The recombinant Smcys10 protein showed binding and agglutinating activity in a Ca2+-dependent manner against bacteria. rSmcys10 treatment upregulated the expression of IL-10, TNF-α and TGF-β in macrophages of turbot and hindered the release of lactate dehydrogenase (LDH) from macrophages after V. anguillarum infection, which confirmed that rSmcys10 reduced the damage to macrophages by V. anguillarum. The NF-κB pathway was suppressed by Smcys10, as demonstrated by dual-luciferase analysis.
CONCLUSIONS: These results indicated that Smcys10 is involved in the host antibacterial immune response.
RESULTS: Cystatin 10 (Smcys10) was cloned from Scophthalmus maximus and encodes a 145 amino acid polypeptide. The results of qRT-PCR showed that Smcys10 exhibited tissue-specific expression patterns, and its expression was significantly higher in the skin than in other tissues. The expression level of Smcys10 was significantly different in the skin, gill, head kidney, spleen and macrophages after Vibrio anguillarum infection, indicating that Smcys10 may play an important role in resistance to V. anguillarum infection. The recombinant Smcys10 protein showed binding and agglutinating activity in a Ca2+-dependent manner against bacteria. rSmcys10 treatment upregulated the expression of IL-10, TNF-α and TGF-β in macrophages of turbot and hindered the release of lactate dehydrogenase (LDH) from macrophages after V. anguillarum infection, which confirmed that rSmcys10 reduced the damage to macrophages by V. anguillarum. The NF-κB pathway was suppressed by Smcys10, as demonstrated by dual-luciferase analysis.
CONCLUSIONS: These results indicated that Smcys10 is involved in the host antibacterial immune response.
摘要：
背景：胱抑素是一种蛋白酶抑制剂，它还调节与炎症相关的基因表达，并在防御和调节中起作用。
结果：胱抑素10（Smcys10）是从大菱杆菌属克隆的，编码145个氨基酸的多肽。qRT-PCR结果显示Smcys10具有组织特异性表达模式,其在皮肤中的表达明显高于其他组织。Smcys10在皮肤中的表达水平有显著差异，吉尔,头肾,anguillarum弧菌感染后的脾脏和巨噬细胞，这表明Smcys10可能在对鳗鱼弧菌感染的抵抗中起重要作用。重组Smcys10蛋白以Ca2依赖性方式对细菌显示出结合和凝集活性。rSmcys10处理上调了大菱头巨噬细胞IL-10，TNF-α和TGF-β的表达，并阻碍了乳酸脱氢酶（LDH）从巨噬细胞感染后，这证实rSmcys10减少了anguillarum对巨噬细胞的损伤。NF-κB途径被Smcys10抑制，如双荧光素酶分析所示。
结论：这些结果表明Smcys10参与宿主抗菌免疫应答。
结果：胱抑素10（Smcys10）是从大菱杆菌属克隆的，编码145个氨基酸的多肽。qRT-PCR结果显示Smcys10具有组织特异性表达模式,其在皮肤中的表达明显高于其他组织。Smcys10在皮肤中的表达水平有显著差异，吉尔,头肾,anguillarum弧菌感染后的脾脏和巨噬细胞，这表明Smcys10可能在对鳗鱼弧菌感染的抵抗中起重要作用。重组Smcys10蛋白以Ca2依赖性方式对细菌显示出结合和凝集活性。rSmcys10处理上调了大菱头巨噬细胞IL-10，TNF-α和TGF-β的表达，并阻碍了乳酸脱氢酶（LDH）从巨噬细胞感染后，这证实rSmcys10减少了anguillarum对巨噬细胞的损伤。NF-κB途径被Smcys10抑制，如双荧光素酶分析所示。
结论：这些结果表明Smcys10参与宿主抗菌免疫应答。
