Haro Hirotaka, Komori Hiromichi, Kato Tsuyoshi, Hara Yasushi, Tagawa Masahiro, Shinomiya Kenichi, Spengler Dan M
Department of Orthopaedic Surgery, Suwa Central Hospital, 4300 Tamagawa, Chino, Nagano 391-8503, Japan.
J Orthop Res. 2005 Mar;23(2):412-9. doi: 10.1016/j.orthres.2004.08.020.
Recently, MMP-7 and MMP-3 have been found to play a crucial role in the natural resorption process of herniated discs. We therefore examined the role of these recombinant human matrix metalloproteinases (rh MMPs) in the treatment of herniated discs.
(a) Surgical samples of herniated disc were cultured in the presence or absence of rh MMPs, and wet weight was measured 24h later. (b) The rh MMPs were administered into normal rabbit intervertebral discs, and after 1 week spine samples were stained with Safranin O. (c) The rh MMPs were administered into canine herniated discs in vivo. Myelography and MRI were performed prior to and 1 week after administration. Spine samples were examined histologically. Whole disc tissue was collected, total protein was extracted, and Western blot analysis was performed.
(a) Proteoglycan degradation was found in MMP-7, MMP-3, and chymopapain-treated samples. MMP-7 and chymopapain-treated samples displayed a significant loss in wet weight (p<0.01). (b) Normal disc tissues after administration of rh MMP-7, MMP-3, and chymopapain showed an extensive loss of Safranin O staining. (c) The rh MMP-7-treated discs had a marked decrease in protruded herniation by MRI. Herniated discs after administration of MMP-7 and chymopapain showed a significant decrease in protruded mass 7 days after administration compared with saline-treated discs when evaluated by myelography (p<0.01). The rh MMP-7-treated discs displayed a clear loss of Safranin O staining in the nucleus pulposus. Proteoglycan expression was barely detectable in disc tissues after MMP-7 administration, whereas obvious expression was obtained in saline-treated or untreated disc tissues.
Exposure to rh MMP-7 resulted in promising proteoglycan loss in human surgical samples, normal rabbit intervertebral discs, and natural canine herniated discs. Administration of rh MMP-7 may facilitate the resorption process of herniated discs.
最近发现基质金属蛋白酶-7（MMP-7）和基质金属蛋白酶-3（MMP-3）在椎间盘突出自然吸收过程中起关键作用。因此，我们研究了这些重组人基质金属蛋白酶（rh MMPs）在治疗椎间盘突出中的作用。
（a）将椎间盘突出手术样本在有或无rh MMPs的情况下培养，24小时后测量湿重。（b）将rh MMPs注入正常兔椎间盘，1周后对脊柱样本进行番红O染色。（c）将rh MMPs体内注入犬椎间盘突出模型。给药前及给药后1周进行脊髓造影和磁共振成像（MRI）检查。对脊柱样本进行组织学检查。收集整个椎间盘组织，提取总蛋白，并进行蛋白质印迹分析。
（a）在MMP-7、MMP-3和木瓜凝乳蛋白酶处理的样本中发现蛋白聚糖降解。MMP-7和木瓜凝乳蛋白酶处理的样本湿重显著降低（p<0.01）。（b）注射rh MMP-7、MMP-3和木瓜凝乳蛋白酶后的正常椎间盘组织番红O染色大量缺失。（c）MRI显示，rh MMP-7处理的椎间盘突出明显减少。脊髓造影评估显示，与盐水处理的椎间盘相比，注射MMP-7和木瓜凝乳蛋白酶后的椎间盘突出肿块在给药7天后显著减小（p<0.01）。rh MMP-7处理的椎间盘髓核番红O染色明显缺失。注射MMP-7后，椎间盘组织中几乎检测不到蛋白聚糖表达，而盐水处理或未处理的椎间盘组织中蛋白聚糖表达明显。
在人类手术样本、正常兔椎间盘和天然犬椎间盘突出模型中，rh MMP-7可导致明显的蛋白聚糖丢失。注射rh MMP-7可能促进椎间盘突出的吸收过程。
