Huang Xiaofei, Niu Man, Sun Tianjing, Li Mo, Jiang Xuheng, Duan Haizhen, Zhang Tianxi, Zhang Ji, Xie Fangke, Song Renjie, Yu Anyong
Department of Emergency, Affiliated Hospital of Zunyi Medical University, 563003 Zunyi, Guizhou, China.
Department of Emergency, Affiliated Hospital of Zunyi Medical University, 563003 Zunyi, Guizhou, China; Department of Emergency, Fourth Hospital of Shijiazhuang, 050035 Shijiazhuang, Hebei, China.
Immunol Lett. 2025 Apr;272:106967. doi: 10.1016/j.imlet.2024.106967. Epub 2024 Dec 26.
The spleen, as the body's largest peripheral immune organ and a crucial source of circulating monocytes, plays a significant role in the acute inflammatory response of spleen-derived macrophages to diseases. Therefore, studying the impact and mechanism of X-ray irradiation on spleen-derived macrophages' inflammatory responses is of great importance.
Extracted and identified mice splenic macrophages were divided into four groups: control group, LPS and ATP co-stimulated non-irradiated group, LPS and ATP co-stimulated group irradiated after 6 h, and LPS and ATP co-stimulated group irradiated after 12 h In the LPS and ATP co-stimulated groups, LPS (1μg/ml) and ATP (5mmol/L) were added to establish an inflammatory model in mice splenic macrophages. The irradiated groups were exposed to a medical linear accelerator (Elekta Synergy), while the non-irradiated groups were placed under the light source for the same duration without irradiation. Protein extraction was performed in each group at 6 h and 12 h post-treatment for subsequent analysis using Western blot, ELISA, RT-qPCR and other relevant methods.
(1) Compared with the non-irradiated group, the cell activity in the groups irradiated for 6 h and 12 h at 8 Gy showed a significant increase (P<0.01). (2) In the LPS and ATP co-stimulated groups irradiated after 6 h and 12 h, the expression of NLRP3 mRNA and protein, IL-18 and IL-1β showed a notable decrease compared to the LPS and ATP co-stimulated non-irradiated group (P<0.05). Additionally, caspase-1 expression of caspase-1 mRNA and protein in the 12 h post-irradiation group also decreased considerably when compared with the LPS and ATP co-stimulated non-irradiated group (P < 0.05). In the groups irradiated after 6 h and 12 h, (3) there was a remarkable decrease in the expression of TWIK mRNA and TWIK2, (4) as well as Gq mRNA and protein, when compared to the LPS and ATP co-stimulated non-irradiated group (P < 0.05). Particularly, the 12 h post-irradiation group exhibited a notable reduction in PKC expression (P < 0.05).
X-ray irradiation is capable of inhibiting the activation of ATP-dependent NLRP3 inflammasomes in splenic macrophages.
脾脏作为人体最大的外周免疫器官以及循环单核细胞的重要来源，在脾脏来源的巨噬细胞对疾病的急性炎症反应中发挥着重要作用。因此，研究X射线照射对脾脏来源巨噬细胞炎症反应的影响及其机制具有重要意义。
将提取并鉴定的小鼠脾巨噬细胞分为四组：对照组、脂多糖（LPS）和三磷酸腺苷（ATP）共刺激未照射组、LPS和ATP共刺激6小时后照射组、LPS和ATP共刺激12小时后照射组。在LPS和ATP共刺激组中，加入LPS（1μg/ml）和ATP（5mmol/L）以建立小鼠脾巨噬细胞炎症模型。照射组接受医用直线加速器（医科达Synergy）照射，未照射组在相同时间置于光源下但不进行照射。在处理后6小时和12小时对每组进行蛋白质提取，随后使用蛋白质印迹法、酶联免疫吸附测定法、逆转录定量聚合酶链反应等相关方法进行分析。
（1）与未照射组相比，8 Gy剂量照射6小时和12小时组的细胞活性显著增加（P<0.01）。（2）在LPS和ATP共刺激6小时和12小时后照射组中，与LPS和ATP共刺激未照射组相比，NLRP3 mRNA和蛋白、白细胞介素18（IL-18）和白细胞介素1β（IL-1β）的表达显著降低（P<0.05）。此外，与LPS和ATP共刺激未照射组相比，照射后12小时组中半胱天冬酶1（caspase-1）mRNA和蛋白的caspase-1表达也显著降低（P<0.05）。在照射6小时和12小时后组中，（3）与LPS和ATP共刺激未照射组相比，TWIK mRNA和TWIK2的表达显著降低，（4）Gq mRNA和蛋白的表达也显著降低（P<0.05）。特别是，照射后12小时组蛋白激酶C（PKC）表达显著降低（P<0.05）。
X射线照射能够抑制脾巨噬细胞中ATP依赖性NLRP3炎性小体的激活。
