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目的观察细颗粒物PM_(2.5)对人角质细胞HaCaT的促炎症作用和过敏反应的影响。方法在雾霾天气和沙尘暴天气下分别收集PM_(2.5),用多次超声水浴法提取PM_(2.5)全颗粒,雾霾天气和沙尘暴天气下收集制备的PM_(2.5)分别记为PM_(2.5)non-dust即PM_(2.5)(N)、PM_(2.5)dust即PM_(2.5)(D)。实验组分为PM_(2.5)(N)、PM_(2.5)(D)及PM_(2.5)+2,4-二硝基氯苯(2,4-dinitrochlorobenzol,DNCB)联合作用组,采用MTT实验、实时荧光定量PCR实验、ELISA检测测定各处理组作用后细胞存活率、相关促炎症因子IL-18、IL-1α、TNF-α表达变化。结果与对照组相比HaCaT细胞经各组处理后的细胞存活率差异无统计学意义(P>0.05)。HaCaT细胞经PM_(2.5)(N)处理后,IL-1α、TNFα的胞内mRNA表达差异无统计学意义(P>0.05);而经PM_(2.5)(D)处理后IL-1α、TNFα的表达明显增加,变化倍数分别上调为2.65倍(P<0.01)、1.69倍(P<0.05);HaCaT细胞经PM_(2.5)(N)、PM_(2.5)(D)处理后IL-18的胞外释放量与对照相比均有显著增加(P<0.01),而胞内表达水平存在差异;且PM_(2.5)预处理可明显拮抗化学致敏剂DNCB导致的IL-18细胞外蛋白含量增加(P<0.05)。颗粒物组分分析显示,TNF-α的产生与PAHs(r=0.929)、WSOC(r=0.905)成正相关;而IL-18和IL-1α的产生与PAHs(r=-0.901,r=-0.864)、WSOC(r=-0.847,r=-0.919)成负相关。结论本研究表明了不同组分PM_(2.5)引发皮肤角质细胞免疫反应的异同;沙尘天气比雾霾天气的PM_(2.5)可能具有更强的促炎症效应;而雾霾天气的PM_(2.5)的致敏反应可能更为持久,颗粒物的致敏反应可能通过TLR2/4等促免疫通路导致。
Objective To observe the effect of PM 2.5 on the proinflammatory effect and allergic reaction of human keratinocytes HaCaT. Methods PM 2.5 (2.5) was collected under foggy and sandstorm conditions, and PM 2.5 particles were extracted by multiple ultrasonic baths. The PM 2.5 collected during the haze and sandstorm were recorded as PM 2.5 ) non-dust that PM_ (2.5) (N), PM_ (2.5) dust PM_ (2.5) (D). The experimental groups consisted of PM 2.5 N, PM 2.5 2.5 and D 2 + 2,4-dinitrochlorobenzol (DNCB) combined with MTT assay Real-time fluorescence quantitative PCR and ELISA were used to measure the cell viability and the expression of IL-18, IL-1α and TNF-α in each treatment group. Results Compared with the control group, the cell viability of HaCaT cells treated by each group showed no significant difference (P> 0.05). There were no significant differences in the intracellular mRNA expressions of IL-1α and TNFα in HaCaT cells treated with PM 2.5 (N) (P> 0.05). However, the expression of IL-1α and TNFα (P <0.01), 1.69-fold (P <0.05) respectively. The expressions of IL-18 in HaCaT cells treated with PM 2.5 and N 2.5 were significantly increased (P <0.01), while the intracellular expression levels were different. PM_ (2.5) pretreatment significantly inhibited the protein content of IL-18 induced by chemosensitizer DNCB Increase (P <0.05). The analysis of the composition of the particles showed that the production of TNF-α was positively correlated with PAHs (r = 0.929) and WSOC (r = 0.905), while the production of IL-18 and IL- ) And WSOC (r = -0.847, r = -0.919). Conclusions This study shows the similarities and differences of skin keratinocyte immune response induced by different components of PM 2.5. The dust weather may have a more pro-inflammatory effect than PM 2.5 in the hazy weather, while PM 2.5 in the haze days ) Sensitization may be more sustained, the sensitization of particulate matter may be caused by TLR2 / 4 and other immune pathways.