吡咯烷二硫代氨基甲酯调节大鼠退变椎间盘IL-6和MMP-9表达及机制研究  

Expression of interleukin-6 and matrix metalloproteinase-9 in degenerated cervical intervertebral disc of rats regulated by pyrrolidine dithiocarbamate and its mechanism

作　　者：漆启华[1] 张遂辉 乐阳[1] 王小华[1] 董谢平[1] 

机构地区：[1]江西省人民医院骨二科,南昌330006

出　　处：《中国矫形外科杂志》2016年第23期2171-2176,共6页Orthopedic Journal of China

基　　金：江西省科技支撑计划资助项目(编号:20121122070047)

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白介素-6(IL-6)、基质金属蛋白酶-9(MMP-9)表达的影响,探讨炎性因子在颈椎间盘退变中的作用及其调控机制。[方法]54只SD大鼠随机分为三组,手术切断大鼠颈后部浅、深肌群构建颈椎间盘退变动物模型。PDTC组:大鼠造模术后腹腔每日注射PDTC直至处死,生理盐水组:大鼠造模术后腹腔每日注射生理盐水直至处死,空白对照组:手术切开大鼠颈后部皮肤后立即缝合并每日腹腔给予生理盐水直至处死。分别于术后10、12、16周处死动物,获取C_(5、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IL-6、MMP-9mRNA表达;Western blot检测椎间盘组织中IL-6、MMP-9蛋白表达。[结果]随时间推移,生理盐水组椎间盘组织结构破坏,纤维环排列紊乱,而PDTC组椎间盘组织结构破坏程度轻。椎间盘组织中都检测到IL-6、MMP-9基因及蛋白表达。空白组各时间点IL-6、MMP-9mRNA蛋白表达无明显变化(P>0.05)。PDTC组及生理盐水组IL-6、MMP-9 mRNA表达量随实验时间的延长而增多,但增速逐渐缓慢。各时间点PDTC组IL-6、MMP-9 mRNA表达量较生理盐水组明显降低(P<0.01)。12周、16周PDTC组IL-6蛋白表达量显著低于生理盐水组(P<0.01),而第10周两组间差异无统计学意义(P>0.05)。各时间点PDTC组MMP-9蛋白表达量较生理盐水组明显降低(P<0.01),以12周时最明显。[结论]大鼠颈椎动力失平衡模型的退变椎间盘组织结构破坏明显,椎间盘中IL-6、MMP-9炎性因子大量表达,NF-κB信号通路参与了IL-6、MMP-9表达变化的调节,PDTC有望成为预防颈椎间盘退变的潜在药物。[ Objective ] To evaluate the effects of pyrrolidine dithiocarbamate （PDTC） on the morphology of cervical inter- vertebral disc and the expression of interleukin - 6 （ IL - 6） and matrix metalloproteinase - 9 （ MMP - 9） in the rat model of cer- vical dynamic imbalance, and to explore the roles of inflammatory cytokines in intervertebral disc degeneration and their regula- tory mechanism. [ Methods] Fifty- four Sprague- Dawley rats were randomly divided into PDTC group, saline group and the blank group. The rat model of intervertebral disc degeneration was established by cutting post - cervical superficial and deep mus- cles of rats. After surgery, the rats in the PDTC group and saline group received daily intraperitoneal injection of PDTC and sa- line solution, respectively, until they were sacrificed. For rats in the blank group, the post - cervical skin incision was sutured right after it was made. After surgery, the rats in the blank group received daily intraperitoneal injection of saline solution until they were sacrificed. Rats were sacrificed at 10, 12, or 16 weeks after surgery. C5.6 discs were harvested and the morphological changes in the intervertebral disc were observed by optical microscopy. Quantitative PCR and Western blot were used to determine the mRNA and protein expression of IL -6 and MMP -9 in intervertebral disc tissue, respectively. [ Results ] The saline group had severe structural destruction of intervertebral disc tissue and disordered fiber ring, while the PDTC group had slight structural destruction of intervertebral disc tissue. The mRNA and protein expression of IL - 6 and MMP - 9 were detected in inter- vertebral disc tissue in each group. In the blank group, there were no significant differences in the mRNA and protein expression of IL - 6 and MMP - 9 between different time points （P 〉 0. 05） . The mRNA expression of IL - 6 and MMP - 9 slowly increased in the PDTC group and the saline group. The PDTC group had significantly lower mRNA expression of IL - 6 and MMP

关 键 词：吡咯烷二硫代氨基甲酯(PDTC) 颈椎动力失衡 白介素-1 基质金属蛋白酶-9 

分 类 号：R681.5[医药卫生—骨科学]