糖皮质激素治疗Duchenne型肌营养不良机制的实验研究

发布时间：2018-11-20 15:39

【摘要】：目的:对应用糖皮质激素(glucocorticoid,GC)治疗Duchenne型肌营养不良(Duchenne muscular dystrophy,DMD)的机制进行探讨。方法:应用DMD动物模型mdx小鼠进行实验,将模型组随机分为3组,Mdx模型组(6周龄)、Mdx安慰剂组(12周龄)、Mdx GC治疗组(12周龄),每组8只,另取正常C57小鼠,6周龄和12周龄各8只,作为正常对照组,分别进行悬吊实验,测量时间,观察治疗前后时间变化;测量血清CK值,了解肌肉损伤情况;对所分离的腓肠肌进行HE染色,在电镜下观察病理变化;检测肌肉中Utrophin蛋白表达状况,应用Western blot进行检测,同时获得核因子-κB(nuclear factor-κB,NF-κB)、I-κB、热休克蛋白70(heat shak protein,HSP70)蛋白表达水平,采用独立样本t检验、方差分析(多重比较)和配对t检验统计方法发现与前者关系所在。结果:GC治疗实验动物出现明显变化,与安慰剂相比具有差异明显:(1)Mdx小鼠与治疗前相比肌力明显好转;治疗前悬吊实验时间为(22±11)s,治疗后为(43.8±24.9)s,P0.001,CK值下降明显,治疗前为(20 230±2 444)U/L,治疗后为(6 457±2 215)U/L,P0.001;(2)Mdx小鼠腓肠肌电镜下进行细胞学观察,发现较前明显好转;(3)Mdx小鼠肌肉组织中HSP70蛋白表达水平明显增加,OD比值由治疗前0.50±0.09上升至治疗后0.75±0.09,P0.001,而Utrophin蛋白表达从0.23±0.03上升至0.42±0.08,P=0.003,NF-κB蛋白表达由治疗前0.33±0.04下降至0.26±0.02,P0.001,治疗前后I-κB蛋白表达由0.21±0.06变至0.18±0.05,P=0.372。结论:应用GC治疗后HSP70表达增强,NF-κB活性下降,Utrophin表达增加,从而使Mdx小鼠的肌肉病变有所延缓,肌力增加。
[Abstract]:Objective: to investigate the mechanism of glucocorticoid (glucocorticoid,GC) in the treatment of Duchenne muscular dystrophy (Duchenne muscular dystrophy,DMD). Methods: DMD animal model mdx mice were used in the experiment. The model group was randomly divided into three groups: Mdx model group (6-week-old), Mdx placebo group) (12-week-old), Mdx GC treatment group), 8 mice in each group, and normal C57 mice. The rats of 6 weeks old and 12 weeks old were used as normal control group. The suspension experiment was carried out, the time was measured, and the changes of time before and after treatment were observed. The serum CK value was measured to understand the muscle injury, the isolated gastrocnemius muscle was stained with HE, and the pathological changes were observed under electron microscope. The expression of Utrophin protein in muscle was detected by Western blot, and the expression level of NF- 魏 B (nuclear factor- 魏 B, NF- 魏 B), I- 魏 B, heat shock protein 70 (heat shak protein,HSP70 was obtained, and the expression of NF-kappa B), I- 魏 B and heat shock protein 70 (heat shak protein,HSP70 was detected by independent sample t test. Analysis of variance (multiple comparisons) and paired t test statistical method found the relationship between the former and the former. Results: there were obvious changes in the experimental animals treated with GC, and there was a significant difference compared with placebo: (1) the muscle strength of Mdx mice was obviously improved compared with that of before treatment; The time of suspension test was (22 卤11) s before treatment and (43.8 卤24.9) sP0.001U / L after treatment. It was (20 230 卤2 444) U / L before treatment and (6 457 卤2 215) U / L P 0.001 after treatment. (2) the cytology of gastrocnemius of Mdx mice was observed under electron microscope. (3) the expression of HSP70 protein in muscle tissue of Mdx mice increased significantly, the ratio of OD increased from 0.50 卤0.09 before treatment to 0.75 卤0.09 P0.001 after treatment, while the expression of Utrophin protein increased from 0.23 卤0.03 to 0.42 卤0.08P0.003. The expression of NF- 魏 B protein decreased from 0.33 卤0.04 before treatment to 0.26 卤0.02p0.001, and the expression of I- 魏 B changed from 0.21 卤0.06 to 0.18 卤0.05Pn0.372 before and after treatment. Conclusion: after GC treatment, the expression of HSP70 increased, the activity of NF- 魏 B decreased, and the expression of Utrophin increased, which delayed the muscle lesion and increased the muscle strength of Mdx mice.
【作者单位】：重庆医科大学附属儿童医院神经内科儿童发育疾病研究教育部重点实验室儿科学重庆市重点实验室儿童发育重大疾病国家国际科技合作基地;
【基金】：重庆市卫生局医学科研资助项目(编号:2012-1-044)
【分类号】：R746.2

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