兔脊索细胞通过与激活的巨噬细胞样 THP-1 细胞共培养调节人纤维环细胞炎症介质的表达。

Rabbit notochordal cells modulate the expression of inflammatory mediators by human annulus fibrosus cells cocultured with activated macrophage-like THP-1 cells.

机构信息

Department of Neurosurgery, Guro Hospital, College of Medicine, Korea University, Guro-gu, Seoul, South Korea.

出版信息

Spine (Phila Pa 1976). 2012 Oct 15;37(22):1856-64. doi: 10.1097/BRS.0b013e3182579434.

DOI:10.1097/BRS.0b013e3182579434

PMID:22472811

Abstract

STUDY DESIGN

We evaluated the influence of rabbit notochordal cells on the expression of inflammatory mediators by human annulus fibrosus (AF) cells cocultured with macrophage-like cells.

OBJECTIVE

To identify the protective effect of rabbit notochordal cells on AF during in vitro inflammation.

SUMMARY OF BACKGROUND DATA

Discogenic pain, which is an important cause of intractable lower back pain, is associated with macrophage-mediated inflammation in the AF. Although rabbit notochordal cells prevent intervertebral disc degeneration, their effects on human AF inflammation remain unknown.

METHODS

Human AF pellets were cocultured for 48 hours with notochordal cell clusters from adult New Zealand White rabbits and phorbol myristate acetate (PMA)-stimulated human macrophage-like THP-1 cells. Conditioned media (CM) from the cocultures were assayed by enzyme-linked immunosorbent assay. The expression of inflammatory mediators in the AF pellets was evaluated by real-time reverse-transcription polymerase chain reaction.

RESULTS

The levels of mRNA for interleukin (IL)-6, IL-8, and inducible nitric oxide synthase (iNOS) in the AF pellets cocultured with notochordal cells and macrophages (hAF[rNC-M]) were significantly lower than those in the AF pellets cultured with macrophages alone (hAF[M]) (P < 0.05). The levels of IL-6 and IL-8 proteins in the CM of hAF(rNC-M) were significantly lower than those in the CM of hAF(M) (P < 0.05). Coculturing with notochordal cells significantly decreased the levels of mRNA for IL-6, IL-8, and iNOS in the macrophage-exposed AF pellets (P < 0.05). After 1 ng/mL IL-1β stimulation, the levels of IL-6 and IL-8 mRNA and the level of IL-8 protein production were significantly decreased in the AF pellets with notochordal cells compared with naïve AF pellets (P < 0.05).

CONCLUSION

In an in vitro coculture system, rabbit notochordal cells reduced the levels of main inflammatory mediators and gene expression in the human AF during inflammation. Therefore, rabbit notochordal cells may constitute an important protective tool against symptomatic disc development.

摘要

研究设计

我们评估了在体外炎症中，兔脊索细胞对与巨噬细胞样细胞共培养的人纤维环（AF）细胞炎症介质表达的影响。

目的

鉴定兔脊索细胞对体外炎症时 AF 的保护作用。

背景资料概要

椎间盘源性疼痛是一种难治性腰痛的重要原因，与 AF 中的巨噬细胞介导的炎症有关。尽管兔脊索细胞可以预防椎间盘退变，但它们对人 AF 炎症的影响尚不清楚。

方法

将人 AF 球与成年新西兰白兔脊索细胞簇和佛波醇肉豆蔻酸酯（PMA）刺激的人巨噬细胞样 THP-1 细胞共培养 48 小时。通过酶联免疫吸附试验检测共培养物的条件培养基（CM）。通过实时逆转录聚合酶链反应评估 AF 球中炎症介质的表达。

结果

与巨噬细胞共培养的兔脊索细胞和巨噬细胞（hAF[rNC-M]）的 AF 球中白细胞介素（IL）-6、IL-8 和诱导型一氧化氮合酶（iNOS）的 mRNA 水平明显低于单独与巨噬细胞培养的 AF 球（hAF[M]）（P <0.05）。hAF[rNC-M]的 CM 中 IL-6 和 IL-8 蛋白水平明显低于 hAF[M]的 CM（P <0.05）。与脊索细胞共培养可显著降低暴露于巨噬细胞的 AF 球中 IL-6、IL-8 和 iNOS 的 mRNA 水平（P <0.05）。在 1ng/mL IL-1β刺激下，与未处理的 AF 球相比，含有脊索细胞的 AF 球中 IL-6 和 IL-8 mRNA 水平和 IL-8 蛋白产生水平均显著降低（P <0.05）。