压力可控型椎间盘退变模型的建立及相关研究

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[目的]建立压力可控型椎间盘退变模型并进行相关研究。[方法]80只成年大鼠随机分为A、B、C、D 4组,分别模拟人类在站立(A组)、坐位直立(B组)、坐位前屈(C组)3种状态下椎间盘内的压力情况,应用恒定加压装置对第9、10尾椎加压,D组为对照组(一般人椎间盘面积为1 250 mm2,大白鼠为2.8 mm2,人在站立位时椎间盘内压力为500 N,坐位为750 N,坐位前屈为1 350 N,换算成大白鼠椎间盘的压力分别是1.12 N,1.68 N,3.08 N)。A、B、C组每日加压4 h,对照组仅在尾椎穿针,不加压;4组分别在7、14 d后取标本进行组织学观察,应用免疫组化和阿利新蓝分析测量胶原纤维(Ⅱ型)和蛋白多糖含量,观察椎间盘退变情况。[结果](1)A组、D组类似,椎间盘组织形态、基质胶原和PG含量和成分无明显变化;(2)C组椎间盘发生软骨细胞变性、坏死,髓核皱缩,纤维环胶原纤维变性、排列紊乱,软骨终板钙化、断裂,胶原纤维排列紊乱等组织形态学改变。髓核和软骨终板中PG总量和Ⅱ型胶原表达下降(P<0.05);(3)B组14 d时形态学未观察到软骨细胞坏死等退变表现,但Ⅱ型胶原以及PG含量比对照组明显减少(P<0.05),退变程度较C组轻。[结论]应用恒定加压装置模拟人类脊柱坐位前屈压力加压大鼠尾椎可迅速建立大鼠椎间盘退变模型。 [Objective] To establish pressure-controlled intervertebral disc degeneration model and conduct related research. [Methods] Eighty adult rats were randomly divided into four groups: A, B, C, and D, which were randomly divided into four groups: group A, group B, group C and group C The pressure on the 9th and 10th caudal vertebrae was fixed by a constant pressure device, and the control group (Group D: control group, average disc area: 1 250 mm2, rat: 2.8 mm2, internal pressure of 500 N, sitting at 750 N and sitting flexion at 1 350 N, converted to a pressure of 1.12 N, 1.68 N, and 3.08 N, respectively, in rats. The rats in group A, B and C were treated with daily pressure for 4 hours, while those in control group were only punctured at the caudal vertebrae and were not pressurized. The specimens were taken for histological observation after 7 and 14 days in each group. Immunohistochemistry and alitame blue The contents of collagen fibers (type Ⅱ) and proteoglycan were measured and the disc degeneration was observed. [Results] (1) There was no significant change in disc tissue morphology, matrix collagen and PG content and composition in group A and group D. (2) Chondrocyte degeneration, necrosis and nucleus pulposus shrinkage occurred in group C Degeneration, disorder, cartilage endplate calcification, fracture, disordered arrangement of collagen fibers and other histological changes. The total amount of PG and collagen Ⅱ in nucleus pulposus and cartilage endplate decreased (P <0.05). (3) No morphological changes such as chondrocyte necrosis were observed in group B at 14th day, but type Ⅱ collagen and PG content Than the control group was significantly reduced (P <0.05), degeneration than C group lighter. [Conclusion] The rat model of intervertebral disc degeneration can be established rapidly by applying a constant pressure device to simulate the flexion pressure of human spine sitting on the caudal vertebra.
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