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Relations of transcription expression of IL-2 with nuclear factor of activated T cells as well as changes of C-Fos and C-Jun after trauma

机译:IL-2转录表达与活化T细胞核因子以及创伤后C-Fos和C-Jun变化的关系

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摘要

Objective: To observe the relations among expression of interleukin-2 ( IL-2 ) in spleen lymphocytes, DNA binding activity of nuclear factor of activated T cells (NFAT) and expression of the partly family members C-Fos, C-Jun after trauma. Methods: A murine closed trauma model was used, animals were sacrificed 6, 12 hours and 1, 4, 7, 10, 14 days, respectively after injury. Spleen lymphocytes were isolated from injured mice and stimulated with concanavalin-A. The culture supernatants were harvested and assayed for IL-2 activity. Total RNA was extracted from spleen lymphocytes and assayed for IL-2 mRNA. Nuclear protein was extracted, and the DNA binding activity of NFAT was measured using an electrophoretic mobility shift assay (EMSA) , the expressions of C-Fos, C-Jun protein determined by Western blot analysis. Results: The expressions of IL-2 activity and IL-2 mRNA in spleen lymphocytes were decreased in injured mice compared with those in control mice, and the most obvious decrease appeared on the 4th day after injury. The DNA binding activity of NFAT decreased gradually and reached the minimum that was only 41 % of the control on the 4th day after injury, which was closely associated with the decline of IL-2 activity and IL-2 mRNA. An decrease in the expression of C-Fos on the 1st and 4th day after injury, trauma had no significant effect on the C-Jun expression. Conclusions: These results suggest that the inhibition of IL-2 expression is partly due to the impairment in the activation of NFAT in injured mice; and the decline in the DNA binding activity of NFAT is partly due to trauma block in the C-Fos expression.
机译:目的:观察创伤后脾淋巴细胞白细胞介素2(IL-2)表达,活化T细胞核因子(NFAT)DNA结合活性与部分家族成员C-Fos,C-Jun表达之间的关系。 。方法:使用小鼠闭合性创伤模型,分别在受伤后6、12小时和1、4、7、10、14天处死动物。从受伤的小鼠中分离出脾淋巴细胞,并用伴刀豆球蛋白A刺激。收获培养物上清液并分析IL-2活性。从脾淋巴细胞中提取总RNA,并测定IL-2 mRNA。提取核蛋白,并通过电泳迁移率分析(EMSA)测量NFAT的DNA结合活性,通过Western印迹分析确定C-Fos,C-Jun蛋白的表达。结果:损伤小鼠脾脏淋巴细胞中IL-2活性和IL-2 mRNA的表达较对照组降低,最明显的下降出现在损伤后第4天。 NFAT的DNA结合活性在受伤后第4天逐渐下降,并达到最低,仅为对照组的41%,这与IL-2活性和IL-2 mRNA的下降密切相关。损伤后第1天和第4天,C-Fos的表达降低,对C-Jun的表达无明显影响。结论:这些结果表明,IL-2表达的抑制部分归因于受损小鼠中NFAT激活的损害。 NFAT的DNA结合活性下降部分是由于C-Fos表达中的创伤受阻。

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