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目的:检测TNF对肥大细胞IL-6、IL-10分泌和组胺释放的影响并探讨其可能的信号转导途径。方法:用不同浓度TNF激发肥大细胞系P815后收集细胞和上清,细胞用细胞激发信号ELISA(CASE)方法检测信号转导通路蛋白ERK、p38、和STAT3磷酸化水平,上清用ELISA检测组胺、IL-6和IL-10的水平。结果:ELISA结果显示TNF促进P815肥大细胞IL-6分泌(P<0.05),但对IL-10分泌和组胺释放无明显影响。ERK信号转导通路抑制剂PD98059和U0126抑制TNF引起的P815肥大细胞IL-6分泌(P<0.05),而p38信号转导通路抑制剂SB203580和STAT3信号转导通路抑制剂AG490不能抑制TNF引起的P815肥大细胞IL-6分泌。CASE结果显示ERK信号转导通路抑制剂PD98059,U0126抑制TNF引起的P815肥大细胞内ERK蛋白磷酸化(P<0.05)而p38信号转导通路抑制剂SB203580和STAT3信号转导通路抑制剂AG490不能抑制TNF引起的P815肥大细胞内p38和STAT3蛋白磷酸化。结论:TNF刺激小鼠肥大细胞P815分泌IL-6可能与ERK信号转导通路的激活有关的。
Objective: To investigate the effects of TNF on the secretion of IL-6, IL-10 and histamine in mast cells and to explore the possible signal transduction pathways. Methods: The cells and supernatant were collected after stimulation of mast cell line P815 with different concentrations of TNF. The phosphorylation levels of ERK, p38 and STAT3 were detected by cell-stimulated signal ELISA (CASE). The supernatant was assayed by ELISA Amine, IL-6 and IL-10 levels. Results: The results of ELISA showed that TNF promoted the secretion of IL-6 in P815 mast cells (P <0.05), but had no significant effect on the secretion of IL-10 and histamine release. ERK signal transduction pathway inhibitors PD98059 and U0126 inhibited TNF-induced IL-6 secretion in P815 mast cells (P <0.05), while p38 signaling pathway inhibitor SB203580 and STAT3 signal transduction pathway inhibitor AG490 did not inhibit TNF-induced P815 mast cells IL-6 secretion. CASE showed that PD98059 and U0126, inhibitors of ERK signal transduction pathway, inhibited phosphorylation of ERK protein in P815 mast cells induced by TNF (P <0.05), whereas p38 signal transduction inhibitor SB203580 and STAT4 signal transduction inhibitor AG490 did not inhibit TNF-induced phosphorylation of p38 and STAT3 proteins in P815 mast cells. CONCLUSION: TNF-stimulated IL-6 secretion by P815 in mouse mast cells may be related to the activation of ERK signal transduction pathway.