中文摘要：

目的利用体外诱导小鼠骨髓细胞获得的高纯度肥大细胞（mast cells，MC），探讨白细胞介素（IL）4参与诱导获得的MC中神经激肽受体1（neurokinin receptor 1，NK－1R）和Fc段受体Ⅰα（FcεRⅠα）的表达情况；建立神经肽P物质（substance P，SP）介导的MC脱颗粒模型，初步探讨SP除通过其特异性受体NK－1R调控MC脱颗粒外，是否还存在FcεRⅠα介导的途径。方法从BALB/c小鼠股骨提取骨髓细胞于RPMI 1640完全培养基中，分不同浓度IL－4诱导组即100 μg/L干细胞因子（stem cell factor，SCF）、15 μg/L IL－3以及0、10、15、20、25 μg/L IL－4中进行体外培养。每周换液，将悬浮细胞移入新的培养体系。倒置显微镜观察并记录细胞生长情况。4周后收集各组细胞及上清液，进行甲苯胺蓝染色、流式细胞术检测CD117鉴定MC，流式细胞术以及蛋白印迹Western blot法分析不同组骨髓MC中FcεRⅠα和NK－1R的表达情况。经鉴定培养成功的骨髓MC中分别加入不同浓度的SP（0、0.01、0.1、1.0、10.0 mg/L）孵育0.5 h，检测上清液和细胞内组胺含量，计算组胺释放率。结果不同浓度IL－4（0、10、15、20、25 μg/L）诱导获得MC表面CD117的阳性率分别为（94.8±1.3）%、（95.7±2.5）%、（94.1±1.3）%、（96.6±1.0）%、（96.6±1.1）%，各组比较差异无统计学意义（F＝8.51, P〉0.05）；FcεRⅠα的阳性率分别为（81.5±2.6）%、（84.2±1.8）%、（91.8±2.0）%、（91.6±1.6）%、（93.0±2.6）%，各组比较差异有统计学意义（F＝15.76, P〈0.05）。不同浓度SP（0、0.01、0.1、1.0、10 mg/L）刺激，20 μg/L IL－4诱导组MC释放组胺含量分别为（20.08±1.50）%、（32.76±2.99）%、（42.90±3.36）%、（50.21±1.29）%、（56.10±3.60）%，对照组分别为（19.37±2.02）%、（19.50±1.50）%、（21.77±1.91）%、（32.00±2.50）%、（33.56±1.25）%，各组间比较差异有统计学意义（P值均〈0.05?

英文摘要：

ObjectiveTo investigate the effect of interleukin-4 （IL-4） stimulation on the expression of FcεRⅠα and NK-1R on mature mast cells（MC） cultured and differentiated from mouse bone marrow stem cells, and then to study if these MC also respond to substance P （SP） both in FcεRⅠα and NK-1R dependent manners. MethodsBone marrow cells were aseptically flushed from BALB/c mouse femurs into complete RPMI 1640, followed by culture with stem cell factor （SCF 100 μg/L）, IL-3 （15 μg/L） and IL-4 （0, 10, 15, 20 and 25 μg/L, respectively）. The culture medium was changed once a week. The morphological changes of culture cells were observed under inverted microscope. After 4 weeks culture, the cells were collected and appraised by toluidine blue staining and flow cytometry. The expressions of surface CD117, FcεRⅠα and NK-1R on these cells were detected by flow cytometry and Western blot. Bone marrow MC were activated with SP （0, 0.01, 0.1, 1.0 and 10 mg/L, respectively） for 30 min. The histamine released into the supernatant and stored in the protoplasm was quantified by enzyme linked immunosorbent assay （ELISA）. The percentage of histamine release was calculated as a percent of total histamine content. ResultsWhen different concentrations of IL-4 （0, 10, 15, 20, 25 μg/L）were added into RPMI 1640, the positive rates of CD117 on MC surface were expressed as （94.8±1.3）%, （95.7±2.5）%, （94.1±1.3）%, （96.6±1.0）%, and （96.6±1.1）%, respectively, and there was no significant difference among these groups （F=8.51, P〉0.05）. The positive rates of FcεRⅠα were expressed as （81.5±2.6）%, （84.2±1.8）%, （91.8±2.0）%, （91.6±1.6）%, and （93.0±2.6）%, respectively, and there was statistically increasing among these groups （F=15.76, P〈0.05）. Then MC were activated by SP （0, 0.01, 0.1, 1.0, 10 mg/L）, histamine from 20 μg/L IL-4 group were released （20.08±1.50）%, （32.76±2.99）%, （42.90±3.36）%、（50.21±1.29）%, （56.10±