高骨桥蛋白水平对双足直立鼠脊柱侧凸发生率和严重程度的影响及动物模型建立相关研究

发布时间：2018-05-21 02:45

本文选题：青少年特发性脊柱侧凸 + 骨桥蛋白　；参考：《第二军医大学》2017年硕士论文

【摘要】：一、研究目的青少年特发性脊柱侧凸(adolescent idiopathic scoliosis,AIS)是一种定义为十岁左右起病,并且没有潜在的致病原因支持其发生及进展的一类脊柱三维畸形,女性较男性多发。目前AIS的病因和发病机制仍然缺乏足够的认识,尽管提出大量的假设,但尚未得到明确的论证。近年来相关研究揭示骨桥蛋白(osteopontin,OPN)可能导致特发性脊柱侧凸发生发展。因此,本课题拟建立高OPN水平双足直立小鼠,观察评估高浓度OPN小鼠和对照组之间脊柱侧凸发生率及严重程度的差异,并收集小鼠血清测量OPN浓度,验证单纯高OPN浓度是否能够直接诱发脊柱侧凸发生。此外,我们还建立了高浓度骨桥蛋白水平诱导的雌性双足直立小鼠,并评估其侧凸发生率及严重程度,论证其是否能作为新型药物干预类AIS动物模型平台。二、研究方法(一)选取体重相近的同周龄C3Heej雄性小鼠120只,随机均分为3个组,其中2组于3周龄时行上肢及鼠尾切除术,并分别腹腔注射高浓度OPN及生理盐水,3组均通过特殊鼠笼诱导小鼠直立体态形成3个月,拍摄X线观察比较各组小鼠脊柱侧凸发生率及严重程度,同时收集小鼠血清,使用ELISA方法测定各组小鼠血清OPN浓度差异。(二)选取体重相近的同周龄C3Heej雄性及雌性小鼠共80只,分为高OPN雌性组、高OPN雄性组、对照组雄性组及对照组雌性组4个组,每组各20只,同法诱导小鼠脊柱直立体态3个月,拍摄X线观察比较各组小鼠脊柱侧凸发生率及严重程度。三、研究结果(一)3个月后高OPN双足鼠组、对照双足鼠组、对照四足鼠组血清OPN浓度分别为207.3±85.1 ng/ml、74.6±20.1 ng/ml、83.7±32.2 ng/ml,高浓度OPN组外周血内OPN含量明显升高。小鼠体长分别为(13.6±1.0)cm、(13.9±0.9)cm、(13.6±0.9)cm。平均Cobb角角度分别为(29.8±6.1)°、(20.1±4.4)°、(16.6±2.9)°,侧凸发生率分别为92.5%、52.1%、12.5%。各组小鼠体长未见明显差异,高OPN双足鼠组侧凸发生率及严重程度较对照组明显加重,差异有统计学意义。(二)高OPN雌性双足鼠组、高OPN雄性双足鼠组、对照组雌性组、对照组雄性组Cobb角分别为(25.8±6.7)°、(20.9±6.8)°、(15.6±3.1)°、(17.1±4.5)°,侧凸发生率分别为90%、80%、40%、45%。高OPN雌性组、高OPN雄性组相比对照组之间,侧凸发生率及严重程度差异有显著性,高OPN雌性组较OPN雄性组脊柱侧凸严重程度更重,差异有统计学意义。四、结论本研究结果在动物模型上证实了高OPN水平可能在青少年特发性脊柱侧凸的发病机制中发挥重要作用:其不仅提高了在双足直立小鼠发生脊柱侧凸的风险,而且也与侧凸曲度的进展密切相关,实验结果证实高OPN是导致AIS发病及进展的重要因素;高浓度OPN水平诱导的雌性双足直立小鼠其侧凸发生率更高,侧凸严重程度较重,其更贴近于人类特发性脊柱侧凸女性高发的实际情况,可作为新型药物干预类特发性脊柱侧凸良好的动物模型平台。
[Abstract]:First, adolescent idiopathic scoliosis (AIS) is a class of spinal three-dimensional deformities defined as the onset of the onset of ten years old and without potential cause for disease. Women are more likely than men. At present, the cause and pathogenesis of AIS are still lacking enough understanding, though A large number of hypotheses have not yet been demonstrated. In recent years, relevant studies have revealed that osteopontin (OPN) may lead to the development of idiopathic scoliosis. Therefore, a high OPN level bipedal erect mouse is proposed to assess the incidence of scoliosis and the severity of scoliosis between high concentration OPN mice and control groups. In addition, the concentration of OPN in the serum of mice was collected to verify whether the pure high OPN concentration could induce scoliosis directly. In addition, we also established a high concentration osteopontin level induced female bipedal erect mouse, and evaluated the incidence and severity of scoliosis, and demonstrated whether it could be used as a new drug intervention AIS animal model. Platform. Two. Study method (1) 120 C3Heej male mice with similar weight of the same week age were selected and divided into 3 groups randomly, of which 2 groups were treated with upper limb and tail excision at 3 weeks of age, and high concentration of OPN and saline were injected into the abdominal cavity respectively. The 3 groups were induced by special squirrel cage to form the erect state for 3 months. The incidence of scoliosis and the severity of scoliosis in mice were collected and the serum levels of OPN were collected by ELISA method. (two) a total of 80 C3Heej male and female mice of the same body weight were selected to be divided into high OPN female group, high OPN male group, the control group male group and the control group of 4 groups, with 20 rats in each group. The incidence of scoliosis and severity of scoliosis in mice were compared for 3 months. Three, the results of the study were (1) high OPN bipedal rats after 3 months. The serum OPN concentration of the control quadruped rat group was 207.3 + 85.1 ng/ml, 74.6 + 20.1 ng/ml, 83.7 + 32.2 ng/ml, and high concentration OPN group peripheral blood. The content of internal OPN increased significantly. The body length of the mice was (13.6 + 1) cm, (13.9 + 0.9) cm and (13.6 + 0.9) cm., respectively (29.8 + 6.1), (20.1 + 4.4) degrees, (16.6 + 2.9) degrees, and the incidence of scoliosis was 92.5%, 52.1%, and 12.5%., respectively. The incidence and severity of scoliosis in the high OPN bipedal rats were more than that of the control group. The difference was statistically significant. (two) high OPN female bipedal rats, high OPN male bipedal rats, control group female group, and control group male group Cobb angle respectively (25.8 + 6.7) degrees, (20.9 + 6.8) degrees, (15.6 + 3.1) degrees, (17.1 + 4.5) degrees, the incidence of scoliosis was 90%, 80%, 40%, high OPN female group, high OPN male group compared to the control group, to the side of the control group, to the side of the control group, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side, to the side of the control group. There is a significant difference in the incidence and severity of convexity. The high OPN female group is more serious than the OPN male scoliosis, and the difference is statistically significant. Four. Conclusion the results of this study suggest that high OPN level in the animal model may play an important role in the pathogenesis of adolescent idiopathic scoliosis: it not only improves the double foot in the bipedal system. The risk of scoliosis in erect mice is related to the risk of scoliosis and is closely related to the progression of scoliosis. Experimental results confirm that high OPN is an important factor leading to the onset and progression of AIS. High concentration of OPN level induced female double foot erect mice have higher incidence of scoliosis and severe scoliosis, which is more close to human idiopathic scoliosis. The practical situation of high incidence can be used as a good animal model platform for new drugs to intervene in idiopathic scoliosis.
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R726.8

【参考文献】