Toll样受体激动剂对小鼠脾脏细胞中SARM蛋白表达的影响

The effect of TLR ligands on expression of SARM in mouse splenocytes

目的探讨多种Toll样受体(toll-like receptor, TLR)激动剂分别刺激小鼠脾脏细胞后对SARM蛋白(sterile-α and armadillo motif-containing protein)表达的影响。方法无菌条件下收集小鼠脾脏并制成单细胞悬液,分别加入TLR1/2激动剂Pam3Cys、TLR2/6激动剂LTA、TLR3激动剂poly(I∶C)、TLR4激动剂LPS或TLR9激动剂CpG,终质量浓度均为20μg/mL,刺激时间分别为2、4、8、20和24 h,然后利用蛋白质印迹法(Western blot)检测SARM的表达。结果小鼠脾脏细胞中SARM的表达,Pam3Cys刺激后4~20 h持续上调,24 h后回落接近初始水平;LTA刺激后8~24 h持续下调;poly(I∶C)刺激后4~20 h持续上调,24 h后回落接近初始水平;LPS刺激后20~24 h持续上调;CpG刺激后24 h开始下调。结论小鼠脾脏细胞中SARM可能参与对TLR信号传导通路的调控。

Objective To investigate the effect of multiple TLR(Toll-like receptor)ligands on SARM protein(sterile-α and armadillo motif-containing protein)expression in mouse splenocytes. Methods Mouse spleens were collected under aseptic condition and prepared into single-cell suspension. Then splenocytes were incubated with different TLR agonist(Pam3 Cys for TLR1/2 agonist, LTA for TLR2/6 agonist, poly(I∶C) for TLR3 agonist, LPS for TLR4 agonist or CpG for TLR9 agonist at a final concentration of 20 μg/mL, for 2, 4, 8, 20 and 24 hours, respectively. The expression of SARM was detected by western blot. Results After Pam3 Cys incubation, the expression of SARM in mouse splenocytes was significantly increased continuously from 4 to 20 hours, fell back close to the initial level until 24 hours. After LTA incubation, the expression of SARM in mouse splenocytes was significantly decreased continuously from 8 to 24 hours. After poly(I∶C) incubation, the expression of SARM in mouse splenocytes was significantly increased continuously from 4 to 20 hours, fell back close to the initial level until 24 hours. After LPS incubation, the expression of SARM in mouse splenocytes was significantly increased continuously from 20 to 24 hours. After CpG incubation, the expression of SARM in mouse splenocytes was significantly decreased until 24 hours. Conclusion SARM may be involved in both positive and negative regulations of TLR signaling pathways.