IL-4对肝片吸虫感染诱导的巨噬细胞中脂肪酸结合蛋白2分布的影响

为阐明小鼠感染肝片吸虫后巨噬细胞中脂肪酸结合蛋白2(FABP2)的分布,采用肝片吸虫囊蚴为感染源,经口分别感染雌性BALB/c野生型(WT)及其IL-4单抗处置小鼠,在运用特异性PCR鉴定成功感染小鼠后获取巨噬细胞,并设立人工巯基醋酸盐诱导巨噬细胞对照组进行体外培养。用荧光定量PCR检测选择性激活巨噬细胞的标记蛋白Relmα、Ym1和 Arginase1以确定其激活状态。采用特异性抗体对所获取的巨噬细胞细胞质染色后用共聚焦显微镜摄取不同时间点从不同小鼠体内获取的巨噬细胞染色后的图片。野生型BALB/c巨噬细胞中Relmα、Ym1和Arginase1均大量表达;IL-4单抗处置BALB/c小鼠和巯基醋酸盐诱导小鼠中巨噬细胞中的Relmα、Ym1和Arginase1的表达量较野生型小鼠中的极显著或显著下降。FABP2在FeMΦ中并不分布于细胞质膜下,而是呈点状广泛分布于细胞质中,其荧光强度在12~24 h间呈上升趋势,在24~48 h间却呈下降趋势。

Effects of IL-4 on Distribution of FABP2 in Fasciola hepatica Elicited Macrophages

Macrophages were acquired from the F.hepatica metacercariae infected IL-4 monoclonal antibody treated and wild type female BALB/c mice, respectively. All F.hepatica elicited macrophages and control macrophages were cultured in vitro. mRNAs from macrophages were extracted for the detection of Relmα, Ym1 and Arginase1 genes using Real-time polymerase chain reaction. Pictures and fluorescence intensity at different time points were acquired with a confocal microscopy and image software respectively after staining with serial purified antibodies. Results showed that levels of Relmα, Ym1 and Arginase1 in F.hepatica elicited macrophages(FeMΦ)and control macrophages (TeMΦ) from IL-4 monoclonal antibody treated mice were significantly decreased comparison to that in wild type mice.FABP2 was detected as spots in the cytoplasm. The fluorescence intensity of FABP2 increased from 12 to 24 h, but decreased afterward to 48 h. The fluorescence intensity of FABP2 in TeMΦ was similar to that in FeMΦ.