黄芪甲苷经由miR-125a-5p/NLRP1轴减轻椎间盘突出髓核细胞损伤  

Astragaloside IV attenuates nucleus pulposus cell injury in herniated discs via the miR-125a-5p/NLRP1 axis

作　　者：王新立[1] 刘汝银[1] 王西彬[1] 岳宗进[1] 许大勇[1] 李云朋[1] WANG Xinli;LIU Ruyin;WANG Xibin;YUE Zongjin;XU Dayong;LI Yunpeng(Department of Spine,Henan Hospital of Traditional Chinese Medicine,Zhengzhou 450003,China)

机构地区：[1]河南省中医院脊柱科,河南郑州450003

出　　处：《沈阳药科大学学报》2024年第7期929-938,共10页Journal of Shenyang Pharmaceutical University

摘　　要：目的在白介素-1β(IL-1β)诱导退变的人髓核细胞中,探究黄芪甲苷对miR-125a-5p及其靶基因介导的信号通路的作用,揭示黄芪甲苷(astragaloside IV,AS-IV)对髓核细胞增殖、凋亡以及炎症反应的影响。方法实时荧光定量PCR(RT-qPCR)检测miR-125a-5p和核苷酸寡聚化结构域(NOD)样受体蛋白1(nucleotide oligomerization domain(NOD)-like receptor protein 1,NLRP1)在椎间盘突出患者髓核组织中的表达,用IL-1β诱导髓核细胞退变,在20、50和80μg·mL^(-1)黄芪甲苷干预浓度下检测miR-125a-5p和NLRP1的表达,在IL-1β和80μg·mL^(-1)黄芪甲苷处理的髓核细胞中单独或共同转染miR-125a-5p模拟物和NLRP1过表达质粒,然后分别检测细胞增殖、凋亡和炎症因子分泌情况,蛋白质免疫印迹(Western blot)检测细胞中信号通路相关蛋白p56和p38的磷酸化水平。结果椎间盘突出(lumbar disc herniation,LDH)患者的髓核组织中miR-125a-5p表达下调,NLRP1表达上调。黄芪甲苷促进IL-1β处理的髓核细胞中miR-125a-5p表达,减少NLRP1表达以及p56和p38蛋白的磷酸化水平。黄芪甲苷促进髓核细胞增殖,减少凋亡和炎症反应。结论黄芪甲苷通过上调miR-125a-5p表达,抑制NLRP1的表达和NF-κB/MAPK信号通路,减少IL-1β诱导的髓核细胞损伤。Objective In human nucleus pulposus cells with IL-1β-induced degeneration,we investigated the effects of astragaloside IV(AS-IV)on signaling pathways mediated by miR-125a-5p and its target genes to reveal the effects of AS-IV on the proliferation,apoptosis,and inflammatory responses of human nucleus pulposus cells.Methods RT-qPCR was used to detect the expressions of miR-125a-5p and Nucleotide oligomerization domain(NOD)-like receptor protein 1(NLRP1)in the nucleus pulposus tissue of patients with disc herniation.IL-1βwas used to induce nucleus pulposus cell degeneration,and the expressions of miR-125a-5p and NLRP1 were detected at the intervention concentrations of AS-IV at 20,50 or 80μg·mL^(-1).The nucleus pulposus cells were treated with 80μg·mL^(-1) AS-IV and IL-1β,and transfected with miR-125a-5p mimic and NLRP1 overexpression plasmid separately or jointly,and then the cell proliferation,apoptosis and secretion of inflammatory factors were detected respectively,Western blotting was used to detect the phosphorylation levels of signal pathway related proteins p56 and p38 in cells.Results MiR-125a-5p expression was down-regulated and NLRP1 expression was upregulated in nucleus pulposus tissues of lumbar disc herniation(LDH)patients.Intervention of IL-1β-treated nucleus pulposus cells with AS-IV promoted miR-125a-5p expression and reduced NLRP1 expression as well as phosphorylation levels of p56 and p38 proteins.AS-IV promoted nucleus pulposus cell proliferation,reduced apoptosis and the secretion of inflammatory factors in cells.Conclusion AS-IV reduces IL-1β-induced damage to nucleus pulposus by up-regulating the expression of miR-125a-5p,inhibiting the expressions of NLRP1 and activation of the NF-κB/MAPK signaling pathway.

关 键 词：黄芪甲苷 人髓核细胞 miR-125a-5p NLRP1 NF-κB/MAPK 信号通路 

分 类 号：R28[医药卫生—中药学] R96[医药卫生—中医学]