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背景:胶原纤维是维持椎间盘正常结构与功能的重要物质,椎间盘中胶原的类型及含量的变化,影响整个椎间盘的生理和病理状态,Ⅰ型及Ⅱ型胶原是椎间盘胶原的主要组分。目的:观察六味地黄丸含药血清对肿瘤坏死因子α致伤兔椎间盘Ⅰ型、Ⅱ型胶原蛋白和基因表达的影响。方法:将15只新西兰白兔带终板的45个L2-L5椎间盘随机分为空白第1天组、空白第14天组、肿瘤坏死因子α组、中药血清组、肿瘤坏死因子α+中药血清组。后两种培养液中分别含5μg/L肿瘤坏死因子α和体积分数10%六味地黄丸血清,培养14d后收集髓核和纤维环观察。结果与结论:RT-PCR和WesternBlot检测结果表明,随着培养时间的延长,纤维环中Ⅰ型胶原及髓核中Ⅱ型胶原蛋白和基因表达明显降低;加入肿瘤坏死因子α可加速这种变化,而含药血清可延缓Ⅰ,Ⅱ型胶原蛋白和基因表达变化。提示六味地黄丸可通过稳定Ⅰ,Ⅱ型胶原蛋白和基因表达,对椎间盘细胞外基质具有保护作用。
BACKGROUND: Collagen fibers are important substances that maintain the normal structure and function of intervertebral discs. The type and content of collagen in the intervertebral discs affect the physiological and pathological conditions of the entire intervertebral disc. Collagen type I and II are the main components of collagen in the disc. Objective: To observe the effect of Liuweidihuangwan serum on expressions of type Ⅰ and type Ⅱ collagen and gene in the intervertebral disc of rabbits induced by tumor necrosis factor α. Methods: 45 L2-L5 intervertebral discs with endplate in 15 New Zealand white rabbits were randomly divided into blank day 1 group, blank day 14 group, tumor necrosis factor α group, traditional Chinese medicine serum group, tumor necrosis factor α + serum group. The latter two kinds of culture medium containing 5μg / L tumor necrosis factor α and 10% volume of Liu Wei Di Huang Wan serum, cultured 14d after the collection of nucleus pulposus and fibrous ring observation. RESULTS AND CONCLUSION: The results of RT-PCR and Western Blot showed that with the prolongation of culture time, the expression of type Ⅱ collagen and type Ⅱ collagen in the annulus fibrosus and the nucleus pulposus were significantly decreased. The addition of tumor necrosis factor α accelerated this change , While serum containing drugs can delay the changes of type Ⅰ and type Ⅱ collagen and gene expression. It is suggested that Liuweidihuangwan could protect the extracellular matrix of the intervertebral disc by stabilizing the expression of type I and type II collagen and gene.