Effect of small interference RNA (siRNA) for ADAMTS5 on intervertebral disc degeneration in the rabbit anular needle-puncture model.
Effect of small interference RNA (siRNA) for ADAMTS5 on intervertebral disc degeneration in the rabbit anular needle-puncture model.
复制标题
DOI：
10.1186/ar2851
复制
发表时间：
2009
影响因子：
4.9
通讯作者：
Kimura T
中科院分区：
文献类型：
--
作者：
Seki S;Asanuma-Abe Y;Masuda K;Kawaguchi Y;Asanuma K;Muehleman C;Iwai A;Kimura T
椎间盘退行性病变的病因尚不清楚。一些研究者已经报道了在退化的人类椎间盘中存在蛋白水解酶，如基质金属蛋白酶（MMP）和ADAMTS（一种具有血栓反应蛋白样重复序列的崩解素和金属蛋白酶）家族。Glasson及其同事最近报道，与野生型小鼠相比，ADAMTS5基因敲除小鼠软骨破坏的严重程度显著降低。本研究的目的是评价注射ADAMTS5小干扰RNA （siRNA）寡核苷酸对兔环针穿刺模型椎间盘退变的抑制作用。用ADAMTS5或对照特异性siRNA寡核苷酸转染兔髓核（NP）细胞。采用实时聚合酶链反应（real-time polymerase chain reaction， PCR）评估siRNA转染对ADAMTS5基因的抑制作用，在单层和海藻酸盐珠培养中，有或没有白细胞介素-1β (IL-1β)刺激。采用兔环针穿刺模型（对照组：n = 8； ADAMTS5组：n = 8）在体内检测siRNA的作用。初始环穿刺1周后，分别注射对照或抗adamts5寡核苷酸（L2/3和L4/5水平各100 μg， 16个盘/组）。评估椎间盘高度、磁共振成像（MRI）（汤普森分类和信号强度）和藏红花素- o染色（组织学分级）。IL-1β处理显著提高NP细胞ADAMTS5 mRNA表达水平（P < 0.01）。在有或没有IL-1β刺激的单层和海藻酸珠培养中，ADAMTS5基因的抑制率均比对照低70%。体内注射抗adamts5寡核苷酸后，MRI评分提高，信号强度增加，组织学分级评分提高，差异均有统计学意义（P < 0.05）。椎间盘高度未见明显变化。单次注射ADAMTS5 siRNA可抑制NP组织的降解，MRI和组织学评分均有显著改善。对siRNA的反应机制可能值得探索，以达到可能的治疗目的。
The etiology of degenerative disc disease is unknown. Several investigators have reported the presence of proteolytic enzymes, such as the matrix metalloproteinase (MMP) and ADAMTS (a disintegrin and metalloprotease with thrombospondin-like repeats) families, in degenerated human discs. Glasson and colleagues recently reported that a significant reduction occurs in the severity of cartilage destruction in ADAMTS5 knockout mice compared with wild-type mice. The purpose of this study was to evaluate the suppressive effects of injections of ADAMTS5 small interference RNA (siRNA) oligonucleotide on intervertebral disc degeneration in the rabbit anular needle-puncture model. Rabbit nucleus pulposus (NP) cells were transfected with siRNA oligonucleotides specific for ADAMTS5 or the control. The suppression of the ADAMTS5 gene by siRNA transfection was assessed by using real-time polymerase chain reaction (PCR), both in monolayer and alginate bead cultures with or without interleukin-1β (IL-1β) stimulation. The effect of siRNA was determined in vivo by using the rabbit anular needle-puncture model (control group: n = 8; ADAMTS5 group: n = 8). One week after the initial anular puncture, the animals received an injection of the control or anti-ADAMTS5 oligonucleotide (100 μg each at the L2/3 and L4/5 level; 16 discs/group). Disc height, magnetic resonance imaging (MRI) (Thompson classification and signal intensity), and safranin-O staining (histologic grade) were assessed. IL-1β treatment significantly increased the ADAMTS5 mRNA level in NP cells (P < 0.01). ADAMTS5 gene suppression was 70% compared with the control oligonucleotide in both monolayer and alginate bead culture with or without stimulation with IL-1β. The injection of anti-ADAMTS5 oligonucleotide in vivo resulted in improved MRI scores with increased signal intensity and improved histologic grade scores with statistical significance (P < 0.05). No significant change in disc height was observed. A single injection of ADAMTS5 siRNA induced the suppression of degradation in NP tissues, as shown by significantly improved MRI and histologic grades. The mechanism of response to siRNA may be worthy of exploration for possible therapeutic purposes.
登录
查看更多内容
影响因子：
15.9
作者：
SANDY, JD;FLANNERY, CR;LOHMANDER, LS
通讯作者：
LOHMANDER, LS
影响因子：
3
作者：
Chiba, K;Andersson, GBJ;Thonar, EJMA
通讯作者：
Thonar, EJMA
影响因子：
4.9
作者：
Le Maitre, Christine L;Hoyland, Judith A;Freemont, Anthony J
通讯作者：
Freemont, Anthony J
影响因子：
4.9
作者：
Le Maitre, Christine Lyn;Hoyland, Judith Alison;Freemont, Anthony J
通讯作者：
Freemont, Anthony J
影响因子：
--
作者：
Majumdar, Manas K.;Askew, Roger;Glasson, Sonya S.
通讯作者：
Glasson, Sonya S.