摘要
目的 探索以自我互补双链腺相关病毒9为载体介导的人胰岛素样生长因子-1 (scAAV9-IGF-1)对SOD1-G93A转基因小鼠抗凋亡通路的作用。方法 采用肌萎缩侧索硬化(ALS)的动物模型即转基因SOD1-G93A突变型及野生型(wild type-SOD1,WT-SOD1)小鼠,在其出生后60 d龄时,雌性同窝阳性SOD1-G93A转基因小鼠采用随机的方法分配到治疗组及溶剂对照组,治疗组全身多点肌肉注射scAAV9-IGF-1,溶剂对照组多点肌肉注射AAV9-GFP,同年龄WT-SOD1作为阴性对照组。在肌肉注射40~50 d后,利用PCR技术检测腰髓中IGF-1含量的变化,同时检测抗凋亡通路因子Bcl-xl、Bcl-2的mRNA含量变化,通过免疫组化染色观察抗凋亡通路因子Bcl-xl、Bcl-2在小鼠腰髓前角神经元中的表达。结果 PCR技术检测显示scAAV9-IGF-1处理后,腰髓中IGF-1的mRNA含量显著高于GFP对照组,抗凋亡通路因子Bclxl、Bcl-2的mRNA含量均高于溶剂对照组(均P<0.05),而与WT组差异无统计学意义。免疫组化染色结果显示,治疗组中抗凋亡通路蛋白Bcl-xl,Bcl-2在小鼠腰髓前角神经元中的表达多于溶剂对照组,并且与WT阴性对照组相当。结论 scAAV9-IGF-1可以激活SOD1-G93A转基因小鼠模型中的抗凋亡通路,其通过上调Bcl-xl,Bcl-2的mRNA水平,进而增加Bcl-xl、Bcl-2的蛋白表达,从而产生抗凋亡的作用。
Objective Explore the effect of human insulin-like growth factor-1(scAAV9-IGF-1) mediated by self-complementary double stranded adenovirus 9 on the anti-apoptotic pathway in SOD1-G93A transgenic mice.Methods Animal models of amyotrophic lateral sclerosis(ALS) were used,including transgenic SOD 1-G93A mutant mice and wild-type(WT-SOD1) mice,which were randomly assigned to the treatment group and solvent control group at 60 days after birth.The treatment group received systemic multi-point intramuscular injection of scAAV9-IGF-1,the solvent control group received multi-point intramuscular injection of AAV9-GFP,and the same age WT-SOD1mice served as the negative control group.After 40-50 days of intramuscular injection,the mRNA content of IGF-1 was measured,and the anti-apoptotic pathway factors Bcl-xl,and Bcl-2 cord were detected using PCR technology in the lumbar spinal of ALS models.The expression of anti-apoptotic pathway proteins Bcl-xl,and Bcl-2 were observed by immunohistochemistry staining in the anterior horn of lumbar spinal cord.Results PCR technology detection showed that after scAAV9-IGF-1 treatment,the mRNA levels of IGF-1 was significantly higher than GFP controls,and the mRNA levels of anti-apoptotic pathway factors Bcl-xl,and Bcl-2 were higher than those in the solvent control group in the lumbar spinal cord(~(all)P <0.05),while there was no statistically significant difference compared to the WT group.The immunohistochemical staining results showed that in the lumbar spinal anterior horn neurons of mice in the treatment group,the expression of anti-apoptotic pathway proteins Bcl-xl,and Bcl-2 was higher than that in the solvent control group,and it was comparable to the WT negative control group.Conclusion ScAAV9-IGF-1 activated the antiapoptotic pathway in the SOD1-G93A transgenic mouse model,which upregulated the mRNA levels of Bcl-xl,and Bcl-2,and increased the protein expression of Bcl-xl,and Bcl-2,therefore resulted in an anti-apoptotic effect.
作者
温迪
吉盈肖
李秋生
陈相
刘亚坤
Wen Di;Ji Yingxiao;Li Qiusheng;Chen Xiang;Liu Yakun(Department of Neurology,the Second Hospital of Hebei Medical University,Shijiazhuang 050000,China)
出处
《脑与神经疾病杂志》
CAS
2024年第2期106-110,共5页
Journal of Brain and Nervous Diseases
基金
河北省自然科学基金青年科学基金项目(H2021206048)。
