中医药导报2025，Vol.31Issue(9)：1-8,23,9.DOI:10.13862/j.cn43-1446/r.2025.09.001
补肾活血汤对椎间盘退变模型大鼠巨噬细胞极化介导炎症-焦亡机制的影响
Effect of Bushen Huoxue Decoction(补肾活血汤)on the Macrophage Polarization-Mediated Inflammation-Pyroptosis Mechanism in Rat Models of Intervertebral Disc Degeneration
摘要
Abstract
Objective:To explore the anti-inflammatory and anti-pyroptosis effects of Bushen Huoxue Decoction(BHD)on a rat model of intervertebral disc degeneration(IDD),based on the interaction mechanism between macrophage polarization response and nucleotide oligomerization domain-like receptor thermal protein domain 3(NLRP3)/cysteinyl aspartate specific proteinase-1(Caspase-1)/interleukin-1β(IL-1β)signaling pathway.Method:Among 44 SPF-grade male SD rats,12 rats were randomly selected as blank control group,and the remaining 32 were the surgical model group.The surgical mode group rats were modeled using a new annulus fibrosus puncture modeling method to establish the IDD rat model.Totally 30 successfully modeled rats were randomly divided into model control group,traditional Chinese medicine group and inhibitor(MCC950)group according to the random number table,with 10 rats in each group.Each group was given corresponding drug intervention for 4 weeks.After the intervention cycle ended,all rats were sacrificed and the caudal intervertebral disc tissues were removed.Hematoxylin-eosin(HE)staining was used to observe the pathological morphology of intervertebral discs and conduct histological grading and scoring.The ultrastructure and pyroptosis of nucleus pulposus cells were observed by transmission electron microscopy(TEM).The level of tumor necrosis factor-α(TNF-α)in intervertebral disc tissue was detected by enzyme-linked immunosorbent assay(ELISA).The positive expression levels of M0-type macrophage marker protein(CD68)and M1-type macrophage marker protein(CD197,NLRP3,Caspase-1 and IL-1β)in intervertebral disc tissues were detected by immunohistochemistry(IHC).The relative expression levels of NLRP3,Caspase-1 and IL-1β proteins in the tissues of intervertebral discs were detected by Western blotting.Result:The morphological structure of the intervertebral disc tissue in the blank control group rats was normal,and the annulus fibrosus was neatly and densely arranged.In the model control group,the intervertebral disc morphology and structure of rats were disordered,and the nucleus pulposus tissue was deformed and disordered in arrangement.There were fissures in the annulus fibrosus in model control group.The intervertebral disc tissue structure was basically intact in traditional Chinese medicine group and MCC950 group.A small amount of swelling and deformation of the nucleus pulposus tissue could be observed,and there was a slight fissure in the annulus fibrosus in traditional Chinese medicine group and MCC950 group.The model control group showed higher histological score than blank control group(P＜0.01).The traditional Chinese medicine group and MCC950 group showed lower histological scores than model control group(P＜0.01).The model control group showed higher number of pyroptosis bodies in the nucleus pulposus cells,than blank control group(P＜0.01).The traditional Chinese medicine group and MCC950 group showed lower number of pyroptosis bodies in the nucleus pulposus cells,than model control group(P＜0.01).The model control group showed higher level of TNF-α in the intervertebral disc tissue,than blank control group(P＜0.01).The traditional Chinese medicine group and MCC950 group showed lower level of TNF-α in the intervertebral disc tissues,than model control group(P＜0.01).The model control group showed higher positive expression levels of CD68,CD197,NLRP3,Caspase-1 and IL-1β proteins in the intervertebral disc tissues,than blank control group(P＜0.01).The traditional Chinese medicine group and MCC950 group showed lower positive expression levels of CD68,CD197,NLRP3,Caspase-1 and IL-1β proteins in the intervertebral disc tissues,than model control group(P＜0.01).The model control group showed higher relative expression levels of NLRP3,Caspase-1 and IL-1βproteins in the intervertebral disc tissues,than blank control group(P＜0.01).The traditional Chinese medicine group and MCC950 group showed lower relative expression levels of NLRP3,Caspase-1 and IL-1β proteins in the intervertebral disc tissues,than model control group(P＜0.01).Conclusion:Bushen Huoxia Decoction may inhibit the NLRP3/Caspase-1/IL-1β pyroptosis pathway mediated by macrophage polarization,thereby inhibiting macrophage infiltration and polarization,reducing the release of pro-inflammatory factors and pyroptosis of nucleus pulposus cells,and alleviating the pathological damage of intervertebral disc tissue in IDD.关键词
椎间盘退变/补肾活血汤/巨噬细胞/炎症/NLRP3/Caspase-1/IL-1β/细胞焦亡/大鼠Key words
intervertebral disc degeneration/Bushen Huoxia Decoction/macrophage/inflammation/NLRP3/Caspase-1/IL-1β/pyroptosis of cells/rat分类
医药卫生引用本文复制引用
宁子柳,苏忱,巴拉提·买提卡斯木,许铮荣,冯帅华,吴官保,肖赛,李泽湘,杨届,谢芳,文哲,肖宇旃..补肾活血汤对椎间盘退变模型大鼠巨噬细胞极化介导炎症-焦亡机制的影响[J].中医药导报,2025,31(9):1-8,23,9.基金项目
湖南省自然科学基金面上项目(2023JJ30365,2022JJ30025) （2023JJ30365,2022JJ30025）
湖南省中医药科研计划项目(A2023016,B2024033,B2024038) （A2023016,B2024033,B2024038）
长沙市自然基金项目(kq2403134) （kq2403134）
湖南中医药大学研究生创新课题(2023CX42) （2023CX42）
