Dombrowski Malcolm E, Olsen Adam S, Vaudreuil Nicholas, Couch Brandon K, Dong Qing, Tucci Michelle, Lee Joon Y, Vo Nam V, Sowa Gwendolyn
Department of Orthopaedic Surgery, Ferguson Laboratory for Orthopaedic Research, University of Pittsburgh, Pittsburgh, PA, USA.
Department of Anesthesiology, University of Mississippi Medical Center, Jackson, MS, USA.
Neurospine. 2020 Mar;17(1):69-76. doi: 10.14245/ns.2040046.023. Epub 2020 Mar 31.
Rabbit annulus fibrosus (AF) cells were exposed to isolated or combined mechanical and inflammatory stress to examine the expression of neuropeptide Y (NPY). This study aims to explore the ability of AF cells to produce NPY in response to mechanical and inflammatory stress.
Lumbar AF cells of 6- to 8-month-old female New Zealand white rabbits were harvested and exposed to combinations of inflammatory (interleukin-1β) and mechanical (6% or 18%) tensile stress using the Flexcell System. NPY concentrations were measured in the media via enzyme-linked immunosorbent assay. The presence of NPY receptor-type 1 (NPY-1R) in AF cells of rabbit intervertebral discs was also analyzed via immunohistochemistry and immunofluorescence.
Exposure to inflammatory stimuli showed a significant increase in the amount of NPY expression compared to control AF cells. Mechanical strain alone did not result in a significant difference in NPY expression. While combined inflammatory and mechanical stress did not demonstrate an increase in NPY expression at low (6%) levels of strain, at 18% strain, there was a large-though not statistically significant-increase in NPY expression under conditions of inflammatory stress. Lastly, immunofluorescence and immunohistochemistry of AF cells and tissue, respectively, demonstrated the presence of NPY-1R.
These findings demonstrate that rabbit AF cells are capable of expressing NPY, and expression is enhanced in response to inflammatory and mechanical stress. Because both inflammatory and mechanical stress contribute to intervertebral disc degeneration (IDD), this observation raises the potential of a mechanistic link between low back pain and IDD.
将兔纤维环(AF)细胞暴露于单独或联合的机械和炎症应激下,以检测神经肽Y(NPY)的表达。本研究旨在探讨AF细胞在机械和炎症应激下产生NPY的能力。
采集6至8月龄雌性新西兰白兔的腰椎AF细胞,使用Flexcell系统将其暴露于炎症(白细胞介素-1β)和机械(6%或18%)拉伸应激的组合中。通过酶联免疫吸附测定法测量培养基中的NPY浓度。还通过免疫组织化学和免疫荧光分析兔椎间盘AF细胞中1型NPY受体(NPY-1R)的存在情况。
与对照AF细胞相比,暴露于炎症刺激下NPY表达量显著增加。单独的机械应变并未导致NPY表达有显著差异。虽然在低(6%)应变水平下,炎症和机械应激联合作用并未显示NPY表达增加,但在18%应变水平下,在炎症应激条件下NPY表达有大幅增加(尽管无统计学意义)。最后,分别对AF细胞和组织进行免疫荧光和免疫组织化学检测,证实了NPY-1R的存在。
这些发现表明兔AF细胞能够表达NPY,并且其表达在炎症和机械应激下会增强。由于炎症和机械应激均促成椎间盘退变(IDD),这一观察结果提示了腰痛与IDD之间存在机制联系的可能性。