防风醇提物对肥大细胞PAR-2及相关细胞因子的影响

Effect of Saposhnikoviae Radix Extract on PAR-2 Expression and Related Cytokine Secretion of Mast Cells

目的:观察防风醇提物对肥大细胞过氧化物酶体增殖因子活化受体-2(PAR-2)及相关细胞因子的影响,探索防风抗过敏的新机制。方法:用胰蛋白酶刺激P815细胞的方法建立肥大细胞脱颗粒模型,设空白组,模型组,防风高、低剂量组(0.02,0.01 g·m L^(-1)),药物作用6 h后,采用ELISA检测细胞上清液中组胺,白细胞介素-4(IL-4),IL^(-1)3水平,Western blot,RTPCR检测PAR-2蛋白及其mRNA的表达。结果:与空白组比较,模型组组胺,IL-4,IL^(-1)3含量及PAR-2蛋白及mRNA的表达明显升高(P<0.05,P<0.01);与模型组比较,防风醇提物在体外抑制肥大细胞组胺,IL-4,IL^(-1)3含量及PAR-2蛋白及mRNA的表达(P<0.05,P<0.01)。结论:防风醇提物可能通过抑制PAR-2表达,阻断肥大细胞脱颗粒,且选择性减少相关细胞因子分泌,继而抑制肥大细胞"瀑布效应",达到抗过敏作用。

Objective： To observe the effect of Saposhnikoviae Radix on protease-activated receptors-2 （PAR-2） expression and the related cytokine secretion of mast cells, and explore new anti-allergic mechanism of Saposhnikoviae Radix. Method： Mast cell degranulation models were established by stimulating P815 cells with trysin in vitro. Blank group, model group, Saposhnikoviae Radix high dose group and low dose group （0.02, 0.01 g · mL^-1） were set up. After 6 h of treatment, ELISA assay was used to detect histamine, interleukin-4 （IL-4） and IL-13 levels in cell supernatant. Western blot and RT-PCR were used to detect PAR-2 protein and mRNA expression of P815 cells. Result： Compared with the blank group, histamine, IL-4, IL-13 levels and PAR-2 protein and mRNA expression were significantly increased in model group （P〈0.05, P〈0.01）. Compared with model group, Saposhnikoviae Radix in vitro significantly inhibited the histamine, IL-4, IL-13 levels and PAR-2 protein and mRNA expression in mast cells （P〈0.05,P〈0.01）. Conclusion： By reducing the expression of PAR-2, Saposhnikoviae Radix extract can inhibit mast cell degranulation, selectively decrease the secretion of related cytokines and then suppress the ＂waterfall effect＂ of mast cells, eventually resulting in anti-allergic effect.