MiRNA338对实验性自身免疫性神经炎模型大鼠的治疗作用

发布时间：2018-05-19 13:12

本文选题：Mi + RNA-338　；参考：《内蒙古民族大学》2017年硕士论文

【摘要】：目的:观察miRNA-338对实验性自身免疫性神经炎(EAN)模型大鼠的治疗作用。方法:30只健康、纯系Lewis雌性大鼠随机分为三组,即为正常对照组(n=10)、miRNA-338治疗组(n=10)、生理盐水治疗组(n=10);MiRNA-338治疗组和生理盐水治疗组大鼠采用P0180-199多肽分别注射双后肢足底皮下诱导EAN模型。免疫第7天时,从miRNA-338治疗组大鼠双侧坐骨神经走行区选取三个相同注射位点,注射携带miRNA-338的慢病毒载体;生理盐水治疗组大鼠相同位置注射相同剂量的生理盐水;三组大鼠从免疫开始记录行为学变化并评分,比较高峰期最高评分,在免疫后第18天对各组大鼠进行神经肌肉动作电位、腓肠肌HE染色、运动终板乙酰胆碱酯酶、甲苯胺蓝染色、坐骨神经电镜组织学及免疫组织化学等指标检测,评估治疗效果。结果:(1)与正常组相比较,miRNA-338治疗组行为学改变接近正常,脚趾握力和行走能力接近正常,高峰期最大平均评分与生理盐水治疗组组对比显著改善(p0.05);(2)与生理盐水治疗组相比较,miRNA-338治疗组电生理检测显示高峰期坐骨神经复合动作电位传导速度VmicroRNA-338治疗组V生理盐水治疗组(p0.05),振幅AmicroRNA-338治疗组A生理盐水治疗组(p0.01);(3)模型动物均出现:神经纤维直径变小、排列疏松、横切面形状不均一,腓肠肌纤维变细、胞浆淡染,以及运动终板乙酰胆碱酯酶数量减少等改变。上述指标,与生理盐水治疗组相比较,miRNA-338治疗组得到明显改善。(4)电镜结果显示,生理盐水治疗组坐骨神经髓鞘出现蜂窝状改变,髓鞘内层与轴索分离剥脱,伴随轴索变细等现象,相对与生理盐水治疗组,miRNA-338治疗组剥脱及空洞处可见细胞修复的组织结构,说明miRNA-338对髓鞘空洞和剥脱有明显的修复作用。(5)免疫组化显示,与生理盐水治疗组比较,miRNA-338治疗组高峰期S100和NF200表达都增高,但S100增高的更为明显,基本接近正常。结论:本实验结果显示:1.miRNA-338能明显改善EAN模型大鼠行为学评分、神经传导功能、组织形态学等指标。2.为临床寻找新的治疗自身免疫炎性疾病提供新思路。
[Abstract]:Aim: to observe the therapeutic effect of miRNA-338 on experimental autoimmune neuritis in rats. Methods Thirty healthy, pure Lewis female rats were randomly divided into three groups: normal control group (n = 10), control group (n = 10) treated with miRNA-338, saline group (n = 10) treated with MiRNA-338 and saline group (n = 30). The EAN model was induced by subcutaneous injection of P0180-199 polypeptide into the foot-floor of both hind limbs. On the 7th day of immunization, three same injection sites were selected from the walking area of bilateral sciatic nerve in the miRNA-338 group, and the lentivirus vector carrying miRNA-338 was injected into the rats of the saline treatment group, and the same dose of normal saline was injected into the rats of the saline treatment group at the same position. The behavioral changes were recorded and scored in the three groups from the beginning of immunity, and the highest score was compared during the peak period. The neuromuscular action potential, HE staining of gastrocnemius muscle and acetylcholinesterase of motor endplate were performed on the 18th day after immunization. Toluidine blue staining, histological and immunohistochemical examination of sciatic nerve were used to evaluate the therapeutic effect. Results compared with the normal group, the behavior changes of miRNA-338 treatment group were close to normal, and the grip strength and walking ability of toes were close to normal. Peak maximum mean score was significantly improved compared with saline treatment group (P 0.05). Compared with saline treatment group, the electrophysiological test of miRNA-338 treatment group showed that the peak period sciatic nerve compound action potential conduction velocity (VmicroRNA-338) treatment group had V birth. Model animals in saline treatment group (P 0.05) and amplitude AmicroRNA-338 group (A normal saline treatment group) all appeared: the diameter of nerve fibers became smaller, The fibers of gastrocnemius became thinner, the cytoplasm was pale, and the number of acetylcholinesterase in motor endplate was decreased. The results of electron microscope showed that the myelin sheath of the sciatic nerve in the saline treatment group was honeycombed, the inner layer of the myelin sheath was separated from the axonal cord, and the axonal cord became thinner. Compared with the normal saline treatment group, the tissue structure of the exfoliation and cavities can be seen in the treated group, indicating that miRNA-338 has obvious repair effect on the cavity of myelin sheath and exfoliation. The expression of S100 and NF200 increased in the peak period of the treatment group compared with that in the saline treatment group, but the expression of S100 was more obvious and close to normal in the control group. Conclusion: the results showed that 1. MiRNA-338 could significantly improve the behavioral score, nerve conduction function and histomorphology of EAN rats. To find a new clinical treatment of autoimmune inflammatory disease to provide a new idea.
【学位授予单位】：内蒙古民族大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R744.5;R-332

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