联合应用BMP-2及细胞分化抑制因子1基因延缓兔腰椎间盘退变的实验研究

发布时间：2018-06-12 19:11

  本文选题：椎间盘退变 + 基因治疗　； 参考：《重庆医科大学》2017年硕士论文

【摘要】：目的 研究兔腰椎间盘髓核内注射慢病毒携带的BMP-2和细胞分化抑制因子1(inhibitor of differentiation 1,Id1)基因能否有效改善椎间盘退变进程。方法 (1)经腹建立兔腰椎间盘细针穿刺退变模型,并在影像学和组织学上加以验证;(2)取3月龄新西兰兔32只,体质量2.0~2.5 kg;以L3/4、L4/5、L5/6为目标椎间盘,经腹细针穿刺法制造椎间盘退变模型。将30只造模成功的实验动物随机分为5组,每组6只;造模后4周于各组目标椎间盘中分别注射Lv-BMP-2病毒(A组)、Lv-BMP-2+Lv-Id1病毒(B组)、Lv-Id1病毒(C组)、Lv-绿色荧光蛋白空载体病毒(D组)及PBS(E组)。于注射后2、4、8周各组分别随机取2只动物行T2-mapping MRI检查获得确切的T2弛豫时间(T2值);之后处死动物取椎间盘组织,应用实时荧光定量PCR、ELISA法检测Ⅱ型胶原及蛋白聚糖的m RNA相对表达量及含量。结果 (1)手术操作本身对兔子的体质量无明显影响,仅一只兔子发现腹壁与肠管轻度部分粘连;术后证实出穿刺的椎间盘为目标椎间盘,其椎间盘高度指数和MRI信号强度呈进行性退变改变;穿刺后4周组织学表现为髓核区域脊索细胞显著减少而纤维细胞增生,纤维环结构紊乱。(2)T2-mapping MRI检查示,注射后0、2周各组间T2值比较差异均无统计学意义(P0.05);注射后4周,A、B组T2值显著高于C、D、E组(P0.05),A、B组间比较差异无统计学意义(P0.05);注射后8周,B组T2值显著高于其余各组(P0.05)。实时荧光定量PCR及ELISA检测示,注射后2、4、8周B组Ⅱ型胶原及蛋白聚糖的m RNA相对表达量及含量均显著高于其余各组(P0.05)。结论 (1)经腹纤维环细针穿刺法,操作简单,目标椎间盘定位准确,手术视野更佳,椎间盘退变程度一致,并发症少;而且在影像学和组织学上符合人腰椎间盘退变情况;(2)联合应用BMP-2和Id1基因在体内实验中能有效延缓兔腰椎间盘退变的进程。
[Abstract]:Objective to investigate whether injecting lentivirus-carrying BMP-2 and 1(inhibitor of differentiation 1 / Id1 gene into the lumbar disc nucleus of rabbits can effectively improve the process of disc degeneration. Methods 1) A fine needle puncture model of lumbar intervertebral disc degeneration was established through abdomen and verified by imaging and histology. 32 New Zealand rabbits of 3 months old with a body mass of 2.0 ~ 2.5 kg were taken as the target disc, and L3 / 4 / 4 / 5 / 5 / 6 of the intervertebral disc was used as the target. The model of intervertebral disc degeneration was established by transabdominal fine needle puncture. Thirty successful experimental animals were randomly divided into 5 groups (6 in each group), and Lv-BMP-2 Lv-ID1 virus group B (Lv-BMP-2 Lv-ID1 virus group B) and PBSE group (group D) were injected with Lv-BMP-2 virus in each group 4 weeks after the model was established, and Lv-BMP-2 Lv-ID1 virus group B was injected into the target disc of each group 4 weeks after the model was established, and Lv-BMP-2 Lv-ID1 virus group B was treated with Lv-BMP-2 Lv-ID1 virus group B, respectively. At 8 weeks after injection, 2 animals in each group were randomly selected for T 2-mapping MRI to obtain the exact T 2 relaxation time and T 2 value. The relative expression and content of mRNA in collagen 鈪，

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