摘要
目的:探究MK2抑制剂MMI-0100对异基因造血干细胞移植(allo-HSCT)后炎症反应的作用及分子机制。方法:建立allo-HSCT小鼠模型。随机将受鼠分为BMT+NaCl组和BMT+MMI-0100组,并于移植后每天分别注射NaCl和MMI-0100。两组分别于d 7、14取材,HE染色股骨石蜡切片并于光学显微镜下观察骨髓腔病理变化,qPCR和Western blot检测促炎性细胞因子IL-1β和IL-18的基因和蛋白表达水平,流式细胞仪检测巨噬细胞分型情况,Western blot检测NLRP3和Caspase-1的表达水平。结果:BMT+MMI-0100组骨髓腔炎性细胞浸润明显减少。Western blot结果显示BMT+MMI-0100组IL-1β和IL-18的蛋白表达水平在d 7、14均低于BMT+NaCl组,且比较差异有统计学意义(均P<0.01)。qPCR结果显示与BMT+NaCl组相比,BMT+MMI-0100组在d 7和14的IL-18基因表达水平显著降低(均P<0.01);BMT+MMI-0100组IL-1β基因表达水平在d 7较BMT+NaCl组降低(P<0.05),但在d 14有所增高且明显高于BMT+NaCl组(P<0.05)。流式细胞术检测结果显示,相较于BMT+NaCl组,BMT+MMI-0100组M1型巨噬细胞表达下降,M1/M2比值亦下降(均P<0.05)。Western blot检测结果表明,BMT+MMI-0100组NLRP3和Caspase-1蛋白表达水平均低于BMT+NaCl组(均P<0.05)。结论:MMI-0100具有改善allo-HSCT后骨髓炎症损伤的作用,且可能通过减少NLRP3表达促进M2极化来发挥作用。
Objective:To investigate the role of MK2 inhibitor MMI-0100 on inflammatory response after allogeneic hematopoietic stem cell transplantation(allo-HSCT)and related mechanisms.Methods:An allo-HSCT mouse model was established.Recipient rats were randomly divided into BMT+NaCl group and BMT+MMI-0100 group,and were injected with NaCl and MMI-0100 every day after transplantation,respectively.Samples of the two groups were collected on d 7 and 14,femur paraffin sections were stained with HE,and pathological changes in the bone marrow cavity were observed under the light microscope.The gene and protein expression levels of pro-inflammatory cytokines IL-1βand IL-18 were detected by qPCR and Western blot.Macrophage typing was detected by flow cytometry.The expression levels of NLRP3 and Caspase-1 were detected by Western blot.Results:Inflammatory cell infiltration in the bone marrow cavity was significantly reduced in the BMT+MMI-0100 group.Western blot results showed that the protein expression levels of IL-1βand IL-18 in the BMT+MMI-0100 group were decreased compared to the BMT+NaCl group on day 7 and day 14(all P<0.01).The qPCR results showed that compared to the BMT+NaCl group,the IL-18 gene expression levels in the BMT+MMI-0100 group were significantly reduced on day 7 and day 14(both P<0.01).In the BMT+MMI-0100 group,the expression level of IL-1βgene decreased on day 7(P<0.05),but increased and was higher than that in the BMT+NaCl group on day 14(P<0.05).Flow cytometry results showed that the expression of M1 macrophages and M1/M2 ratio decreased in the BMT+MMI-0100 group compared to BMT+NaCl group(all P<0.05).Western blot results showed that the protein expression levels of NLRP3 and Caspase-1 in the BMT+MMI-0100 group were lower than those in the BMT+NaCl group(all P<0.05).Conclusion:MMI-0100 can ameliorate bone marrow inflammatory injury after allo-HSCT and may act by reducing NLRP3 expression to promote M2 polarization.
作者
王兆慧
龙博
王玉寒
刘志婷
徐紫洁
丁爽
WANG Zhao-Hui;LONG Bo;WANG Yu-Han;LIU Zhi-Ting;XU Zi-Jie;DING Shuang(School of Medical Technology,Xuzhou Medical University;Xuzhou Center for Disease Control and Prevention;Department ofClinical Laboratory Examination,The Affiliated Hospital of Xuzhou Medical University,Xuzhou 221000,Jiangsu Province,China)
出处
《中国实验血液学杂志》
北大核心
2025年第5期1453-1460,共8页
Journal of Experimental Hematology
基金
国家自然科学基金(81900106)。
关键词
异基因造血干细胞移植
炎症
MK2
NLRP3
allogeneic hematopoietic stem cell transplantation
inflammation
MK2
NLRP3
