Katsuno Raijiro, Hasegawa Toru, Iwashina Toru, Sakai Daisuke, Mikawa Yoshihiro, Mochida Joji
Department of Orthopedic Surgery, Kawasaki Medical College, Kurashiki, Okayama, Japan.
Spine (Phila Pa 1976). 2008 Apr 15;33(8):845-9. doi: 10.1097/BRS.0b013e31816b4685.
A study of age-related effects on nitric oxide (NO) and cytokine production in cocultured rat nucleus pulposus (NP) cells and macrophages.
To evaluate the effects of age on NO and cytokine production in an in vitro model of cocultured NP cells and macrophages.
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.
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.
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.
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.
一项关于年龄对共培养的大鼠髓核（NP）细胞和巨噬细胞中一氧化氮（NO）及细胞因子产生影响的研究。
在共培养NP细胞和巨噬细胞的体外模型中评估年龄对NO及细胞因子产生的影响。
众所周知，腰椎间盘突出症的临床特征随年龄而异。临床特征的年龄相关差异与免疫化学因子（如NO和炎性细胞因子）之间的关系尚未确立。
使用雄性Sprague Dawley大鼠（n = 45），包括来自3个组（3周龄、12周龄和32周龄）的15只动物。NP细胞和渗出的腹腔巨噬细胞在无血清培养基中共培养。使用Griess法在2小时、24小时、48小时和72小时测量NO水平。培养7天后，评估包括组织金属蛋白酶抑制剂-1、干扰素-γ（IFN-γ）和白细胞介素-10（IL-10）在内的细胞因子的产生。
共培养物中的NO水平随年龄增加。在共培养组中，3周龄组的组织金属蛋白酶抑制剂-1和IFN-γ水平在统计学上高于12周龄和32周龄组。3周龄组的IL-10水平在统计学上低于12周龄和32周龄组。
共培养物中的NO水平随年龄增加，这表明炎症反应随年龄增加。本研究显示了以IFN-γ和IL-10水平为代表的年龄相关细胞因子失衡。应激和衰老被认为会影响细胞外基质并改变免疫反应。年轻大鼠的NP细胞具有较高的细胞介导免疫活性，而老年大鼠具有较高的体液免疫活性。这些结果表明年龄会影响NP细胞的免疫反应。需要进一步研究以阐明这种新观察到现象的机制并将这些发现应用于临床。
