白细胞介素-37 通过核因子-κB 通路影响 PDZ 结构域包含蛋白 1 从而促进痛风发病。
Interleukin-37 contributes to the pathogenesis of gout by affecting PDZ domain-containing 1 protein through the nuclear factor-kappa B pathway.
机构信息
Department of Rheumatology and Immunology, Changhai Hospital, Affiliated to Naval Military Medical University, Shanghai, China.
出版信息
J Int Med Res. 2020 Sep;48(9):300060520948717. doi: 10.1177/0300060520948717.
OBJECTIVE
Our objective was to explore the molecular pathogenesis of the onset of gout and the mechanism underlying the effect of interleukin (IL)-37 on PDZ domain-containing 1 (PDZK1) protein through the nuclear factor-κB signaling pathway.
METHODS
Real-time PCR and western blotting were used to detect expression of PDZK1 mRNA and protein, respectively, in the HK-2 cell line. The inhibitors pyrrolidine dithiocarbamate (PDTC) and wortmannin were added to HK-2 cells stimulated by IL-37, and changes in PDZK1 protein were detected by western blotting.
RESULTS
Based on our previous research, we used 10 µmol/L PDTC. We detected no significant change in at the mRNA level among the IL-37, PDTC+IL-37, and wortmannin+IL-37 groups. With increasing IL-37 concentration, the protein level of PDZK1 increased. After adding wortmannin, the protein level of PDZK1 increased with increasing concentration of IL-37, albeit not significantly, and the level of PDZK1 remained lower than that with IL-37 alone. After adding PDTC, the protein level of PDZK1 showed a trend to decrease with increasing concentrations of IL-37 up to 40 ng/mL. The immunofluorescence results supported the western blot results.
CONCLUSIONS
IL-37 can affect protein expression of PDZK1, but not at the translational level, in the pathogenesis of gout.
目的
本研究旨在探讨痛风发病的分子发病机制，以及白细胞介素（IL）-37 通过核因子-κB 信号通路对 PDZ 结构域包含蛋白 1（PDZK1）蛋白的影响及其作用机制。
方法
采用实时荧光定量 PCR 和 Western blot 法分别检测 IL-37 刺激 HK-2 细胞后 PDZK1mRNA 和蛋白的表达。采用核因子-κB 信号通路抑制剂吡咯烷二硫代氨基甲酸盐（PDTC）和渥曼青霉素（wortmannin）预处理 HK-2 细胞，Western blot 法检测 PDZK1 蛋白的变化。
结果
在前期研究的基础上，选用 10 μmol/L PDTC 作为抑制剂，IL-37、PDTC+IL-37、wortmannin+IL-37 三组细胞 PDZK1mRNA 水平比较差异无统计学意义。随着 IL-37 浓度的增加，PDZK1 蛋白表达逐渐增加，当加入 wortmannin 后，PDZK1 蛋白表达随 IL-37 浓度的增加而逐渐增加，但差异无统计学意义，且低于 IL-37 单独作用组；加入 PDTC 后，PDZK1 蛋白表达随 IL-37 浓度的增加呈逐渐下降趋势，在 40 ng/mL 时达最低值。免疫荧光结果与 Western blot 结果一致。
结论
IL-37 可能通过影响核因子-κB 信号通路，在痛风发病机制中影响 PDZK1 蛋白的表达，但不影响其翻译水平。
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