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目的探讨李斯特菌溶血素LLO诱导THP-1细胞产生IL-1β的分子机制。方法将THP-1细胞随机分为Syk处理组、未处理组及阴性对照组。Western blot检测不同LM菌株感染巨噬细胞不同时相点Syk的磷酸化水平;免疫荧光法观察细胞内炎症复合体的关键蛋白组分ASC-speck的形成情况;实时荧光定量PCR法检测白细胞介素(IL)-1β表达。结果 WT和△hly∶∶hly菌株感染THP-1细胞0 min、15 min、30 min时,Syk磷酸化水平均较△hly明显升高;Syk处理组胞内ASC-speck阳性的细胞百分比明显低于未处理组(P<0.01),△hly菌株感染组的ASC-speck阳性细胞百分比也明显低于WT、hly菌株感染组(P<0.01);Syk处理组产生的IL-1β明显低于未处理组,并且△hly菌株感染THP-1细胞产生的IL-1β较WT和△hly∶∶hly LM感染组明显降低(P<0.05)。结论 Syk介导的信号通路参与调控LM感染过程中LLO介导的炎症复合体的形成。
Objective To investigate the molecular mechanism of LLO-induced IL-1β production in THP-1 cells. Methods THP-1 cells were randomly divided into Syk treatment group, untreated group and negative control group. Western blot was used to detect the phosphorylation of Syk at different time points in macrophages infected with different strains of LM. The formation of ASC-speck, a key protein component of intracellular inflammatory complex, was observed by immunofluorescence assay. The levels of interleukin (IL) -1β expression. Results The Syk phosphorylation levels in THP-1 cells infected with WT and △ hly::hly strains were significantly higher than those in △ hly cells at 0, 15 and 30 min, respectively. The percentage of ASC-speck-positive cells in Syk-treated group was significantly lower The percentage of ASC-speck positive cells in the untreated group (P <0.01) was also significantly lower than that in the WT and hly infected groups (P <0.01); the IL-1β production in the Syk-treated group was significantly lower than that in the untreated group Compared with WT and △ hly:: hly LM groups, the IL-1β production of TH-1 cells infected by △ hly strain was significantly decreased (P <0.05). Conclusion Syk-mediated signaling pathway is involved in the regulation of LLO-mediated inflammatory complex formation during LM infection.