烟酰胺对椎间盘Ⅰ、Ⅱ型胶原的调控作用  

Regulation of Niacinamide on Intervertebral Disc Type Ⅰ and Ⅱ Collagen in vitro

作　　者：徐蔚蔚[1] 邵增务[1] 郭兵[1] 杨述华[1] 俞旭东[1] 谢卯[1] 熊蠡茗[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院骨科,武汉430022

出　　处：《中国中医骨伤科杂志》2008年第6期21-25,共5页Chinese Journal of Traditional Medical Traumatology & Orthopedics

基　　金：湖北省自然科学基金资助项目(No2004ABA193)

摘　　要：目的:考察烟酰胺对白介素-1β诱导的体外兔腰椎间盘退变模型中Ⅰ、Ⅱ型胶原的保护作用,并初步探讨其机制。方法:构建兔腰椎间盘组织凝胶培养模型,将其分为正常对照组,烟酰胺对照组(加入0.5mg/ml烟酰胺),退变组(加入10ng/mlIL-1β)和3个烟酰胺治疗组(加入IL-1β的同时,并分别加入0.5mg/ml,0.25mg/ml和0.05mg/ml烟酰胺)。培养3天及1周时取各组髓核和纤维环分别行Ⅰ、Ⅱ型胶原基因RT-PCR检测,培养2周时行Ⅰ、Ⅱ型胶原、iN-OS、TGF-β1及Caspase-3免疫组化检测。结果:①RT-PCR显示3天及1周时,各治疗组Ⅰ、Ⅱ型胶原表达均强于退变组,烟酰胺的保护作用随浓度增加而增强。②Ⅰ、Ⅱ型胶原免疫组化显示,各治疗组椎间盘正常结构和胶原含量均优于退变组,以大剂量烟酰胺治疗组最显著。③iNOS、Caspase-3免疫组化显示,治疗组阳性细胞染色率均低于退变组(P<0.01)。TGF-β1免疫组化显示,治疗组阳性细胞染色率高于退变组(P<0.01)。结论:烟酰胺能够抑制IL-1β诱发的椎间盘Ⅰ、Ⅱ型胶原表达及含量的下降,这种保护机制与其抑制iNOS、Caspase-3表达和维持TGF-β1表达的作用有关。Objective： To study the effect of niacinamide on type Ⅰ and Ⅱ collagens in intervertebral disc （IVD） degeneration induced by interleukin-1β （IL-1β） in vitro and to investigate the possible mechanism. Methods：Chiba＇s IVD culture model was adopted in this study. IVDs from adult Japanese white rabbits were randomly divided into 6 groups ： normal control, niacinamide group （0.5mg/ml niacinamide）, degeneration group （10ng/ml IL-1β） and treatment group （both IL- 1β and 0.5mg/ml,0.25mg/ml, 0.05mg/ml niacinamide, respectively）. After 3 days and 1 week of culture, the IVDs were collected for type Ⅰ and Ⅱ collagens reverse transcription polymerase chain reaction （RT--PCR）. Two weeks later, IVDs were collected for immunohistochemistry staining for type Ⅰ and Ⅱ collagens, inducible nitric oxide synthase （iNOS）, transforming growth factor-β1 （TGF-β1 ） and easpase- 3. Results： ① RT- PCR showed that the expressions of both type Ⅰ and Ⅱ collagens in treatment group were significantly higher than that in degeneration group and the effect was stronger with denser doses of niacinamide. ②Immunohistochemistry staining of type Ⅰ and Ⅱ collagens demonstrated that the structure of IVDs and contents of collagens were better reversed in treatment group as compared with those in degeneration group. ③Positive- staining of iNOS and caspase- 3 in treatment group was significantly lower while TGF-β1 evidently higher than those in degeneration group （P〈0.01）. Conclasion： Niacinamide can effectively inhibit the decreased expression of type Ⅰ and Ⅱ collagens in IVD induced by IL-1β through suppressing the expression of iNOS and caspase-3 and sustaining the expression of TGF-β1.

关 键 词：椎间盘 烟酰胺 Ⅰ型胶原 Ⅱ型胶原 白细胞介素-1Β 

分 类 号：R-33[医药卫生] R681.53