Age-related effects of cocultured rat nucleus pulposus cells and macrophages on nitric oxide production and cytokine imbalance
Age-related effects of cocultured rat nucleus pulposus cells and macrophages on nitric oxide production and cytokine imbalance
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DOI：
10.1097/brs.0b013e31816b4685
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发表时间：
2008-04-15
期刊：
影响因子：
3
通讯作者：
Mochida, Joji
中科院分区：
文献类型：
--
作者：
Katsuno, Raijiro;Hasegawa, Toru;Mochida, Joji
研究设计。研究年龄对大鼠髓核细胞和巨噬细胞一氧化氮（NO）和细胞因子产生的影响。在体外共培养NP细胞和巨噬细胞模型中评估年龄对NO和细胞因子产生的影响。背景资料摘要。众所周知，腰椎间盘突出症的临床特征随年龄的不同而不同。年龄相关的临床特征差异与免疫化学因子（如NO和炎症细胞因子）之间的关系尚未确定。选用雄性Sprague Dawley大鼠（n = 45），其中15只动物分为3周龄、12周龄和32周龄3组。NP细胞和渗出的腹腔巨噬细胞在无血清培养基中共培养。采用Griess法在2、24、48和72小时测量NO水平。培养7天后，检测细胞因子的产生，包括组织抑制剂金属蛋白酶-1、干扰素- γ (ifn - γ)和白细胞介素-10 （IL-10）。共培养一氧化氮水平随年龄增长而升高。共培养组3周龄组织抑制剂金属蛋白酶-1、ifn - γ水平均高于12、32周龄。3周龄大鼠IL-10水平低于12、32周龄大鼠。共培养物NO水平随着年龄的增长而增加，表明炎症反应随着年龄的增长而增加。这项研究显示了与年龄相关的细胞因子失衡，以ifn - γ和IL-10的水平为代表。应激和衰老被认为会影响细胞外基质并改变免疫反应。年轻大鼠NP细胞具有较高的细胞免疫活性，而老年大鼠具有较高的体液免疫活性。这些结果表明，年龄影响了NP细胞的免疫反应。需要进一步的研究来阐明这一新发现的机制并将这些发现应用于临床。
Study Design. A study of age-related effects on nitric oxide (NO) and cytokine production in cocultured rat nucleus pulposus (NP) cells and macrophages.Objective. To evaluate the effects of age on NO and cytokine production in an in vitro model of cocultured NP cells and macrophages.Summary of Background Data. It is well known that the clinical characteristics of lumbar disc herniation differ with age. The relationship between age-related differences in clinical features and immuno-chemical factors, such as NO and inflammatory cytokines, has not been established.Methods. Male Sprague Dawley rats (n = 45), including 15 animals from 3 groups (3-, 12-, and 32-weeks old), were used. NP cells and exudated peritoneal macrophages were cocultured in serum-free media. NO levels were measured at 2-, 24-, 48-, and 72 hours using the Griess method. After 7 days of culture, the production of cytokines, including tissue inhibitor metalloproteinase-1, interferon-gamma (IFN-gamma), and interleukin-10 (IL-10) were evaluated.Results. NO levels of coculture increased with age. In the coculture groups, tissue inhibitor metalloproteinase-1 and IFN-gamma level of 3 weeks old were statistically higher than 12 and 32 weeks old. IL-10 level of 3 weeks old was statistically lower than 12 and 32 weeks old.Conclusion. NO levels of cocultures increased with age that suggests inflammatory reactions increase with age. This study showed an age-related cytokine imbalance, as represented by levels of IFN-gamma and IL-10. Stress and aging are thought to affect the extracellular matrix and change the immunologic response. Younger rat NP cells had higher cell-mediated immunity activity, while the older rat had higher humoral immunity activity. These results demonstrate that age affects the immunologic response attributable to NP cells. Further studies are needed to elucidate the mechanism of this newly observed occurrence and to apply these findings clinically.