内耳注入携带IL-4基因的骨髓间充质干细胞对豚鼠免疫性内耳病基因治疗的研究

发布时间：2018-01-19 21:52

  本文关键词： 骨髓间充质干细胞 白细胞介素 慢病毒 免疫性内耳病 基因治疗　出处：《听力学及言语疾病杂志》2014年05期 　论文类型：期刊论文

【摘要】：目的研究内耳局部植入白细胞介素4(IL-4)基因修饰骨髓间充质干细胞(bone-marrow mesenchymal stem cells,BMSCs)对免疫性内耳病豚鼠内耳病理损伤和听功能的调节与治疗作用。方法采用钥孔嘁血蓝蛋白(KLH)抗原在已致敏的豚鼠圆窗龛局部免疫,制成豚鼠免疫性内耳病模型55只,分为五组:A组(BMSCs载体组)、B组(BMSCs空载体对照组)、C组(重组慢病毒IL-4基因组)、D组(慢病毒空载体对照组)及E组(模拟手术对照组),每组11只,各组均将相应的悬液20μl[BMSCs悬液中含BMSCs(1.5~2.0)×106,慢病毒浓缩液浓度为0.5×108 pfu]经鼓阶开窗植入内耳。采用免疫荧光组织化学试验法、免疫酶组织化学试验法观察IL-4基因产物在内耳组织结构中的分布和表达情况,酶联免疫吸附试验观察血清抗KLH特异性抗体水平,ABR测试听功能变化。结果与免疫前相比,免疫后A、B、C组ABR波Ⅲ阈值不同程度降低,A组阈值降低更明显,差异有统计学意义(P0.05);免疫组织化学染色显示BMSCs荧光反应阳性细胞主要分布在鼓阶、前庭阶;内耳光镜观察A、B、C组仅在鼓阶内有絮状物,注射部位有少量红细胞和白细胞,D、E组可见不同程度的膜迷路积水,螺旋神经节和蜗轴小血管周围有单个核细胞浸润。结论重组IL-4基因的慢病毒载体可在体外成功转染BMSCs,经鼓阶途径植入内耳后可在内耳迁移并产生基因产物IL-4,可明显减轻免疫性内耳病动物的内耳免疫炎性反应和听功能损伤,而BMSCs可以作为细胞载体把携带有目的基因的治疗因子迁移到损伤部位而发挥治疗作用。
[Abstract]:Objective to study the bone marrow mesenchymal stem cells modified with interleukin-4 (IL-4) gene implanted into the inner ear. The regulation and therapeutic effect of BMSCs on the pathological injury and auditory function of the inner ear of guinea pigs with immune internal ear disease. Methods the key-hole liming hemocyanin (KLHH) antigen was used to immunize the sensitized guinea pig round window niche. Fifty-five guinea pigs with immune internal ear disease were divided into five groups: group A, group B, group B, group B, group C (recombinant lentivirus IL-4 genome). Group D (lentivirus empty vector control group) and group E (simulated operation control group, 11 rats in each group), each group will have the corresponding suspension of 20 渭 l. [BMSCs suspensions contained BMSCs (1. 5 + 2. 0) 脳 10 ~ (6). The concentration of lentivirus concentrate was 0. 5 脳 10 ~ 8 pfu] and the inner ear was implanted with a tympanic window. Immunofluorescence histochemical assay was used. The distribution and expression of IL-4 gene products in the inner ear were observed by immunohistochemistry, and the levels of specific antibodies against KLH in serum were observed by Elisa. Results compared with before immunization, the threshold of ABR wave 鈪，

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