高场强电磁脉冲对Jurkat细胞IFN-γ和IL-4蛋白表达的定量分析

来源 :中国体视学与图像分析 | 被引量 : 0次 | 上传用户:gedebao
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目的 :研究电磁脉冲 (EMP)辐照对白血病肿瘤细胞株Jurkat细胞分泌细胞因子IFN -γ和IL - 4的影响 ,探讨EMP对Jurkat细胞损伤机制。方法 :Jurkat细胞经EMP照射后即刻、1h、 6h、 12h、 2 4h、 4 8h、 72h共 7个时相点 ,细胞悬液甩片 ,4 %多聚甲醛 4℃固定 ,自然晾干 ,进行SP法的IFN -γ和IL - 4蛋白免疫组化染色。在光镜 10× 4 0倍视野下 ,应用CMIAS-H图像分析仪对阳性细胞的图像分析参数积分光密度值 (IOD)进行统计分析。结果 :免疫组化结果表明 ,场强 6× 10 4 V/m的电磁脉冲能使Jurkat细胞IFN -r表达持续性减少 ,差异非常显著 (P <0 0 1) ,而IL - 4变化不明显 (P >0 0 5 )。结论 :EMP辐照后Th1型的细胞因子IFN-γ明显减少 ,而Th2型的细胞因子IL - 4未见显著变化 ,提示 :EMP辐照后机体中受到影响的主要是细胞免疫 ,而体液免疫功能未受到明显影响 Objective: To investigate the effects of EMP irradiation on cytokine IFN-γ and IL-4 secreted by leukemia cell line Jurkat and explore the mechanism of EMP injury in Jurkat cells. Methods: Jurkat cells were irradiated with EMP immediately after 7h, 1h, 6h, 12h, 24h, 48h, 72h. The cell suspensions were fixed with 4% paraformaldehyde at 4 ℃ and allowed to air dry. SP immunohistochemical staining of IFN-γ and IL-4 protein. The statistical analysis of the integral optical density (IOD) value of the image analysis parameters of the positive cells was carried out by using the CMIAS-H image analyzer under the light microscope of 10 × 40 times. Results: The results of immunohistochemistry showed that the electromagnetic radiation with field intensity of 6 × 10 4 V / m decreased the expression of IFN-γ in Jurkat cells continuously (P <0.01), while the change of IL - 4 was not obvious (P> 0 0 5). CONCLUSION: Th1-type cytokine IFN-γ is significantly decreased after EMP irradiation, while there is no significant change in Th2 type cytokine IL-4, suggesting that cellular immunity is mainly affected by EMP irradiation, while humoral immunity Function has not been significantly affected
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