【作者】 朱翔；

【导师】 任永信；

【作者基本信息】 南京医科大学 ， 外科学（骨外）， 2017， 硕士

【摘要】 [目的]研究抑制氧化应激及NF-κ B信号通路在延缓因bmi-1基因缺失引起的椎间盘退变中的作用及机制。[方法]选取4周龄野生型和bmi-1纯合子小鼠,共分4组。A组为4周龄野生型小鼠,经正常饮食饮水喂养;B组为正常喂养的4周龄bmi-1 KO小鼠,正常喂养。C组为4周龄大的bmi-1 KO小鼠,饮水中加入抗氧化剂NAC,使喂养浓度达到1mmol/L;D组为4周龄bmi-1 KO小鼠,予以腹腔注射NF-κ B抑制剂IMD-0354,连续注射一周,每天注射一次,每次注射量为20mg/kg。ABC三组小鼠饲养一个月后取材,D组小鼠腹腔注射一周后取材。通过HE组织学染色对各组小鼠椎间盘的形态学改变进行观察,通过番红固绿染色观察椎间盘蛋白多糖含量。Western-blot方法检测各组小鼠椎间盘氧化应激、NF-κ B活性、分解代谢等相关指标的表达差异。[结果]本研究中HE染色及番红固绿染色结果显示:bmi-1/-小鼠髓核形态发生改变,周围的纤维环排列较野生型小鼠更紊乱,细胞外基质(聚蛋白多糖)表达量较野生型的高。经NAC、IMD处理后的bmi-1/-小鼠椎间盘的形态学改变有所改善。蛋白质印记技术结果显示:bmi-1-/-小鼠椎间盘内氧化应激水平明显高于野生型小鼠,基质降解酶相关指标蛋白表达水平明显上升,经NAC、IMD处理后的bmi-1-/-小、鼠椎间盘内氧化应激水平以及基质降解酶蛋白的表达均有明显下降。[结论]bmi-1基因缺失可以加速小鼠椎间盘的分解代谢,引起小鼠体内椎间盘退变。抑制氧化应激或抑制NF-κ B信号通路能纠正bmi-1缺失引起的椎间盘退变。本研究为临床上延缓和治疗椎间盘退变提供了新的靶点和方向。更多还原

【Abstract】 [Objective]The purpose of this study is to investigate the role of Oxidative Stress and the NF-kappa B pathway in alleviating the intervetebral disc degeneration induced by bmi-1-knockout.[Methods]We design 4 groups for our study,8 WT mice or bmi-1-/-mice are in each group.Mice in group A are WT mice feed with normal food,bmi-1-/-mice feed with normal food in group B,bmi-1-/-mice feed with NAC in group C.bmi-1-/-mice treatment with intraperitoneal injection with IDM-0354 in group D.Samples of group A,B and C were obtained after one month,while samples of group D after one week.We observed the morphological changes of disc in mice by Hematoxylin-Eosin staining and Saffron staining.Proteins and RNA were extraction to evaluate the indicators of Oxidative Stress,NF-κB and catabolism.[Results]Histological staining showed that morphology of NP changes in bmi-1-/-mice,the arrangement of AF surrounding is more disordered than the wild-type mice,the expression of ECM(proteoglycan)was higher than that of wild type mice.Morphological changes have improved in bmi-1-/-mice after treatment with NAC or IMD.Results of protein imprinting technology show that the level of oxidative stress,as well as the expression of matrix degrading enzymes in the intervertebral disc of bmi-1-/-mice,was higher than that in wild type mice.The level of oxidative stress and the expression of matrix degrading enzymes decreased significantly in bmi-1-/-mice after treatment with NAC or IMD.[Conclusion]Deletion of Bmi-1 gene is an important factor to accelerate the degeneration of intervertebral disc.Inhibition of oxidative stress or NF-kappa B signaling pathway can correct Bmi-1-deficiency-induced disc degeneration.This study provides a new target for delaying or treating the occurrence and development of disc degeneration.更多还原