Objective To explore the effect of osthole on mast cells and expression of STAT5 gene and protein in them. Methods Passive cutaneous anaphylaxis was made in mice to copy eczema model, then the allergic mast cells were separated, and the ovalbumin was used to induce allergy of mast cells. Different concentrations of osthole were used to intervene the sensitized mast cells.Then the sensitized mast cells were divided into blank control group, osthole high-dose group and low-dose group.At the end of the experiment, morphology of the mast cells was detected by immunofluorescence technology.MTT assay was used to detect the influence of drugs on mast cells proliferation. RT-PCR and Western blotting were used to detect the expression of STAT5 gene and protein. Results As compared with blank control group, the number of mast cells in the osthole groups was significantly reduced, cells and nuclei obviously shrank, even apoptosis of some cells could be observed; the inhibition rate of mast cells in osthole groups was significantly increased in concentration-dependent manner ( P<0. 01 ) . As compared with blank control group (2.16±0.57), gene expression of STAT5 was significantly decreased in osthole high-dose group (0.59±0.12) and low-dose group (0.82±0.13) (P<0.01).The protein expression of STAT5 was also significantly decreased in osthole high-dose group and low-dose group as compared with blank control group (P<0.01). Conclusion Osthole can inhibit the proliferation of sensitized mast cells, and reduce the expression of STAT5 gene and protein.%目的 探讨蛇床子素对小鼠湿疹肥大细胞及其信号转导和转录活化蛋白5( STAT5)基因和蛋白表达的影响.方法 采用被动皮肤变态反应复制小鼠湿疹模型,致敏后分离腹腔肥大细胞,采用卵蛋白诱发肥大细胞过敏,致敏后的肥大细胞分为空白对照组、蛇床子素大剂量组及小剂量组. 药物干预24 h后,采用免疫荧光法检测3组肥大细胞形态,噻唑蓝法检测药物对肥大细胞增殖的影响,RT-PCR及Western blot法检测各组细胞中STAT5基因及蛋白的表达水平. 结果与空白对照组比较,给药组肥大细胞数量明显减少,细胞体积明显缩小,细胞核体积缩小,部分细胞凋亡;细胞抑制率随药物浓度显著上升(P<0.01);与空白对照组(2.16±0.57)比较,蛇床子素大剂量组(0.59±0.12)和小剂量组(0.82±0.13)STAT5基因表达水平均有极显著降低( P<0.01);与空白对照组比较,蛇床子素大剂量组及小剂量组STAT5蛋白表达水平均有极显著降低( P<0.01). 结论 蛇床子素可抑制致敏肥大细胞的增殖,并下调STAT5基因和蛋白的表达.
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