防风醇提物对肥大细胞PAR-2及相关细胞因子的影响

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(P0.05,P0.01);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(P0.05,P0.01)。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。

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

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.