慢病毒介导的RNA干扰下调IL-6抑制脊髓顿挫损伤后Caspase-3表达

Change of Caspase-3 Expression in Spinal Cord Contusion after Using Lentivirus Inhibited the Expression of IL-6

目的通过慢病毒介导的RNA技术干扰IL-6的表达,探究脊髓顿挫损伤后Caspase-3的表达变化。方法Allen′s打击法制备脊髓顿挫伤(SCC)模型,BBB评分确认造模成功;免疫组织化学和Q-PCR技术定位和定量测定IL-6和Caspase-3的表达变化;Gene MANIA网站预测IL-6与Caspase-3的联系;慢病毒介导的RNA干扰技术构建IL-6干扰模型后,Q-PCR技术定量测定Caspase-3的表达变化。结果 SCC后SD大鼠BBB评分显著低于假手术组(P<0.05);IHC结果显示:IL-6和Caspase-3均在在脊髓白质胶质细胞表达;Q-PCR结果显示:IL-6在损伤后12h表达显著增加,3d、28d表达持续下降;Caspase-3在损伤后12h、3d、28d表达持续下降。生物信息学分析结果:IL-6和Caspase-3在大鼠体内存在共定位关系。干扰组Caspase-3表达比空载组显著降低(P<0.05)。结论在SCC后,抑制IL-6表达可降低Caspase-3表达水平。

Objective This study aimed to explore the change of Caspase-3expression in spinal cord contusion after using lentivirus inhibited the expression of IL-6.Methods The model of spinal cord contusion was prepared by Allen′s method and affirmed the success of making model by BBB score.The expressions of IL-6and Caspase-3were detected by immunohistochemistry technique and quantitative polymerase chain reaction（Q-PCR）.Then,the relationship between IL-6and Caspase-3was forecasted by the method Gene MAIN bioinformatics.Finally,the caspase-3gene expressive was detected by the Q-PCR in spinal cord after interfering in the expression of IL-6.Results After spinal cord injury（SCC）,the score of SD rats is lower than the Sham group（P〈0.05）.In early term,after SCC,the gene expressive quantities of IL-6and Caspase-3became higher obviously（P〈0.05）.And in late term,the gene expressive quantities of IL-6and Caspase-3became lower evidently（P〈0.05）.They both expressed at gliocytes in white matter.After interfering in the expression of IL-6,the gene expressive quantities of Caspase-3is lower distinctly（P〈0.05）.Conclusion After SCC,IL-6could stimulate the expression of Caspase-3.