摘要
目的：观察1，25-二羟基维生素D3[1，25（OH）2D3]对屋尘螨抗原（Dermatophagoides pteronyssinus， Der p）直接作用的P815肥大细胞Toll样受体4（toll-like receptor 4，TLR4）表达及白细胞介素-4（interleukin-4， IL-4）分泌的影响，探讨1，25（OH）2D3在肥大细胞免疫调节中的作用。方法用不同浓度Der p、1，25（OH）2D3单独或联合激发P815细胞，收集激发细胞和上清。用real-time PCR及Westernblot法检测P815细胞TLR4表达，ELISA法检测上清中IL-4浓度。结果（1）P815细胞表达TLR4；不同浓度Der p均可上调P815细胞TLR4表达，且36 h组TLR4表达有明显浓度依赖性上调；（2） Der p可促进IL-4释放（P＜0．05）；（3）不同浓度1，25（OH）2D3单独激发对P815细胞TLR4表达及IL-4分泌均无明显影响（P＞0．05）；（4）10－8 mol／L 1，25（OH）2D3可增强Der p对P815细胞TLR4表达的促进作用（P＜0．01），1，25（OH）2D3可抑制Der p引起的IL-4的分泌（P＜0．05或P＜0．01），且1，25（OH）2D3浓度越高，抑制作用越强。结论（1）尘螨抗原可通过上调肥大细胞TLR4表达，增加IL-4分泌，促进炎症及过敏反应的发生。（2）1，25（OH）2D3对尘螨抗原引起的肥大细胞免疫应答的干预作用可能是通过抑制IL-4的合成和分泌，从而抑制Th2型免疫应答。
Objective To investigate the effects of Dermatophagoides pteronyssinus （ Der p） on the expression of TLR4 and IL-4 in P815 mast cells and to further analyze the immunoregulatory effects of 1,25-（OH）2D3 on Der p treated P815 mast cells.Methods Different concentrations of Der p and 1,25-（ OH） 2 D3 were used alone or in combination to stimulate P815 mast cells.The supernatants of the stimulated cell culture were analyzed by enzyme-linked immunosorbent assay （ ELISA） for the detection of IL-4.The stimulated cells were collected and analyzed by real-time PCR and Western blot assays for the detection of TLR4atmRNAandproteinlevels,respectively.Results （1）TLR4expressionwasdetectedinP815 cells.The expression of TLR4 was enhanced in P815 cells treated with various concentrations of Der p.A significant dose-dependent up-regulation of TLR4 was observed in P815 cells after incubation with Der p for 36 h.（2） Der p promoted the release of IL-4 in P815 cells （P〈0.05）.（3） No significant differences with the expression of TLR4 and IL-4 were observed among 1,25（OH）2D3 treatment groups as compared with the control group （P〉0.05）.（4） 10-8 mol/L of 1,25（OH）2D3 promoted the Der p-induced expression of TLR4 in P815 cells （P〈0.01）.However, 1,25（OH）2D3 inhibited the release of IL-4 in a dose-dependent manner（P〈0.05orP〈0.01）.Conclusion （1）Derpcouldpromotetheinflammationandallergicreac-tion through up-regulating TLR4 and IL-4 in mast cells.（2） The possible mechanism for the inhibitory of 1, 25（OH）2D3 on Der p-induced immune responses was due to the suppression of Th2-type immune responses through inhibiting the synthesis and secretion of IL-4 in mast cells.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2015年第3期188-193,共6页
Chinese Journal of Microbiology and Immunology
基金
浙江省自然科学基金资助项目（LY13H010004）
