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脾脏酪氨酸激酶(SYK)是一种非受体型酪氨酸激酶(non-receptor tyrosine kinase, NRTK)。在哺乳动物中,SYK是重要的细胞信号通路接头分子,其在免疫细胞活化,胞外刺激的信号转导和病原体识别等多种生命过程中扮演重要角色,然而在鱼类中却鲜有报道。本研究以香鱼(Plecoglossus altivelis)为研究对象,探讨了其SYK(PaSYK)在应对病原微生物感染及在活化免疫信号通路中的作用。我们克隆获得了香鱼SYK的cDNA序列,并利用多重序列比对、构建进化树等生物信息学方法分析PaSYK的进化地位;采用实时荧光定量PCR(RT-qPCR)方法分析PaSYK在健康组织及鳗弧菌(Vibrio anguillarum)感染后的表达变化;亚细胞定位检测PaSYK在HEK293T细胞的分布情况;在香鱼头肾单核/巨噬细胞(Monocyte/macrophage,MO/MΦ)中过表达PaSYK,研究其对炎症细胞因子表达的调控作用并在HEK293T细胞及香鱼头肾MO/MΦ中研究PaSYK对MAPK信号通路的活化作用。多重序列比对结果显示,SYK的蛋白质序列和结构域在进化过程中是高度保守的。系统进化树结果表明, PaSYK与河鳟的亲缘关系密切。RT-qPCR结果表明, SYK基因mRNA在检测的所有组织中均有表达,鳗弧菌感染后免疫相关组织中PaSYK表达量均上调。此外,PaSYK能显著诱导香鱼头肾MO/MΦ中促炎因子TNF-α和IL-1β的表达,但对抑炎因子TGF-β和IL-10的表达起到略微的抑制作用。Western blot结果证明PaSYK可以激活MAPK信号通路。综上所述, SYK分子在进化上高度保守,香鱼SYK与其哺乳动物中的同源物具有保守的功能,均能活化MAPK信号通路并在炎症反应中发挥作用。 相似文献
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Effects of carboxymethyl-chitosan with different molecular weights on wound healing were investigated in vivo and in vitro.A second degree burn model was performed in rats and the accelerative effects of carboxymethyl-chitosan on wound repair were observed.Contents of transforming growth factor(TGF)-β1,interleukin(IL)-6 and matrix metalloproteinase(MMP)-1 in wounds were determined by enzyme-linked immunosorbent assay(ELISA).In vitro study evaluated the influence of carboxy-methyl-chitosan on cytokines secretion of fibroblasts and macrophages.In vivo results showed that carboxymethyl-chitosan effec-tively accelerated the wound healing process in burned rats(P<0.05).Levels of TGF-β1,IL-6 and MMP-1 in carboxymethyl-chitosan groups were significantly elevated,compared with control group(P<0.05).In vitro results indicated that carboxymethyl-chitosan significantly promoted the proliferation of fibroblasts and stimulated its IL-8 and IL-10 secretion at different incubation time,but it did not affect collagen secretion of fibroblasts.Carboxymethyl-chitosan enhanced phagocytosis ability of macrophages,and in-creased its tumor necrosis factor(TNF)-α secretion.In conclusion,carboxymethyl-chitosan promoted wound healing by activating macrophages,accelerating fibroblasts growth and exerting considerable effects on the secretion of a series of cytokines. 相似文献
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为了探讨日粮中添加不同剂量的海带粗多糖对斜带石斑鱼(Epinephelus coioides)血清指标的影响,作者选取规格一致的健康斜带石斑鱼120尾(平均体质量为90 g±2.6 g,平均体长为15 cm±2.25 cm),均匀分为4组,每组设两个重复,每个重复15尾。对照组日粮中不添加海带粗多糖,试验0.5%组、1.0%组、1.5%组日粮中分别添加0.5%、1.0%、1.5%的海带粗多糖。经过48 d的喂养后,测定各组斜带石斑鱼血清中白介素2(Interleukin-2,IL-2)、白介素6(Interleukin-6,IL-6)、干扰素α(Interferon-α,IFN-α)、总蛋白(Total Protein,TP)、尿素(UREA)、肌酐(CREA)、碱性磷酸酶(Alkaline phosphatase,ALP)、谷草转氨酶(Aspartate aminotransferase,AST)、谷丙转氨酶(Alanine aminotransferase,ALT)、溶菌酶(Lysozyme,LZM)、过氧化物酶(Catalase,CAT)、超氧化物歧化酶(Superoxide dismutase,SOD)、补体3(Complement3,C3)、补体4(Complement 4,C4)等水平。结果表明,海带粗多糖能够不同程度地提高血清中IL-2、IL-6、IFN-α、TP水平,及LZM、CAT和SOD酶活性;降低C3、C4含量,及UREA、CREA水平和ALP酶活性;对AST和ALT酶活性影响不大。在3种添加比中,以0.5%的海带粗多糖添加剂量效果最明显。 相似文献
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