吡咯烷二硫代氨基甲酯抑制TNF-α和MMP-9表达延缓大鼠颈椎间盘退变及机制研究

摘要

目的 观察吡咯烷二硫代氨基甲酯(PDTC)对大鼠颈椎动力失衡模型颈椎间盘组织形态及肿瘤坏死因子α(TNF-α)、基质金属蛋白酶9(MMP-9)表达的影响,探讨PDTC抑制颈椎间盘退变作用及其机制.方法 54只SD大鼠随机分为3组,切断大鼠颈后部浅、深肌群构建颈椎间盘退变动物模型.PDTC给药组(A组):造模术后腹腔每日注射PDTC溶液.模型组(B组):造模术后腹腔每日注射生理盐水.假手术组(C组):手术切开大鼠颈后部皮肤后立即缝合并每日腹腔注射生理盐水.术后10、12、16周分批处死动物,获取C5/6椎间盘组织,光镜下观察椎间盘形态改变,q-PCR检测椎间盘组织中TNF-α mRNA、MMP-9 mRNA表达;Western blot检测椎间盘组织中TNF-α、MMP-9蛋白表达变化.结果 PDTC给药组(A组)椎间盘结构破坏减轻,纤维环结构排列有序.模型组(B组)TNF-α、MMP-9 基因表达量随实验时间的延长而增多,早期增速较快,后期逐渐缓慢,在12周达高峰,而蛋白表达量在术后10周即达到较高水平,与12周、16周时间点比较无明显差异(P>0.05).各时间点PDTC给药组TNF-α mRNA、MMP-9 mRNA及蛋白表达量较模型组(B组)明显降低(P0.05) in blank group (group C) at each time point.The expressions of IL-6, MMP-9 mRNA increased with time in group B, but the amount increased fast firstly, and slow lately, reaching peak in 12 weeks.The expression of TNF-α, MMP-9 protein became steady in group B from 10 weeks compared with other time points(P>0.05).TNF-α, MMP-9 genes and proteins expression decreased obviously in PDTC group (group A) compared with saline group (group B) (P<0.01) at each time point, but higher than blank group C(P<0.01) at each time point.Conclusions TNF-α and MMP-9 are important inflammatory factors involved in rat cervical disc degeneration, PDTC relieves the degeneration of rat cervical disc by reducing the expression of TNF-α and MMP-9 via disturbing the NF-κB signal pathway probably, and PDTC may become potential medicine for disc degeneration.