退变椎间盘纤维环破裂后髓核免疫微环境的变化  

Changes of immune microenvironment of nucleus pulposus caused by annulus fibrosus rupture in intervertebral disc degeneration

作　　者：白进玉 陈浩[1] 常文举 蒋凤仙[1] 周晓中[1] 田波 单冰晨[1] Bai Jinyu;Chen Hao;Chang Wenju;Jiang Fengxian;Zhou Xiaozhong;Tian Bo;Shan Bingchen(Department of Orthopedics,the Second Affiliated Hospital of Soochow University,Suzhou 215004,China)

机构地区：[1]苏州大学附属第二医院骨科,苏州215004

出　　处：《中华实验外科杂志》2023年第6期1083-1086,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81873995、82172425、82102611)。

摘　　要：目的:探究椎间盘退变(IDD)中纤维环破裂引起的免疫微环境变化。方法:选取2020年9月至2021年7月在苏州大学附属第二医院骨科诊断为椎间盘突出症行手术治疗的患者,根据纤维环破裂与否,分为纤维环完整组(AI)和纤维环破裂组(AR),收集髓核标本。采用流式细胞仪、实时反转录聚合酶链反应(RT-PCR)和酶联免疫吸附法(ELISA)检测组织样本炎性因子表达水平;免疫组织化学染色观察免疫细胞CD68、F4/80、白细胞介素(IL)-1β的表达。结果:流式细胞术分析显示AR组巨噬细胞(37.20±0.64比14.50±0.58,t=45.30,P<0.01)和T细胞比例较AI组明显增多[(22.10±0.67)%比(4.06±0.05)%,t=46.20,P<0.01]。在mRNA表达水平上,AR组TNF-α表达倍数高于AI组(8.73±1.56比1.00±0.04,t=8.57,P<0.01),AR组IL-1β表达倍数高于AR组(5.68±0.70比1.00±0.02,t=11.55,P<0.01),AR组IL-4表达倍数高于AI组(2.33±0.77比1.00±0.05,t=3.01,P<0.05),AR组IL-6表达倍数高于AI组(17.00±1.81比1.00±0.06,t=8.83,P<0.01),差异有统计学意义。在炎症因子水平上,AR组TNF-α高于AI组(12.20±4.56比5.55±3.74,t=2.66,P<0.05)和AR组IL-1β高于AI组(10.60±1.58比6.32±2.61,t=3.41,P<0.05)。免疫组织化学染色显示AR组中CD68、F4/80、IL-1β表达阳性的细胞明显增多。结论:IDD进程中纤维环破裂加剧髓核组织免疫炎症反应,且影响椎间盘退变的病理生理过程。Objective To investigate the changes of immune microenvironment caused by annulus fibrosus rupture in intervertebral disc degeneration(IDD).Methods Patients diagnosed with intervertebral disc herniation and underwent surgical treatment in the orthopedics department of the second affiliated hospital of soochow university from september 2020 to july 2021 were enrolled.According to the annulus rupture or not,patients were divided into annulus intact(AI)group and annulus rupture(AR)group.The nucleus pulposus were collected and then flow cytometry,real-time reverse transcriptase-polymerase chain reaction(RT-PCR)and enzyme-linked immunosorbent assay(ELISA)to detect the levels of inflammatory factors.Immunohistochemical was used to observe the expression of CD68,F4/80 and interleukin(IL)-1βin immune cells.Results Flow cytometry analysis showed that the ratio of macrophages and T cells(37.20±0.64 vs.14.5±0.58,t=45.30,P<0.01)in AR group was higher than that in AI group[(22.10±0.67)%vs.(4.06±0.05)%,t=46.20,P<0.01].At the mRNA expression level,AR expression of TNF alpha multiples than AI group(8.73±1.56 vs.1.00±0.04,t=8.57,P<0.01),IL-1βexpression ratio in AR group was higher than that of AR group(5.68±0.70 vs.1.00±0.02,t=11.55,P<0.01),The expression ratio of IL-4 in AR group was higher than that in AI group(2.33±0.77 vs.1.00±0.05,t=3.01,P<0.05),and the expression ratio of IL-6 in AR group was higher than that in AI group(17.00±1.81 vs.1.00±0.06,t=8.83,P<0.01),and the difference was statistically significant.TNF-αin AR group was higher than that in AI group(12.20±4.56 vs.5.55±3.74,t=2.66,P<0.05)and IL-1βin AR group was higher than that in AI group(10.60±1.58 vs.6.32±2.61,t=3.41,P<0.05).Immunohistochemical staining showed that cells with positive expressions of CD68,F4/80 and IL-1βincreased significantly in AR group.Conclusion The annulus fibrosus rupture during IDD exacerbates the immune inflammatory response in nucleus pulposus and affects the pathophysiological process of disc degeneration.

关 键 词：椎间盘退变 纤维环破裂 免疫微环境 炎性反应 

分 类 号：R681.53[医药卫生—骨科学]