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目的筛选偏头痛大鼠三叉神经节(TG)中调控IL-1β的微小RNA(miRNA)。方法34只SD大鼠行矢状窦旁硬脑膜外置管后随机分为假手术组(不用药,n=13)、注射致炎剂(IS)10μl/d1次(IS 1d组,n=4)、2次(IS 2d组,n=4)和3次组(IS 3d组,n=13)。Western blot检测TG中IL-1β表达。提取假手术组及IS 3d组大鼠TG的RNA进行miRNA芯片检测;软件预测靶作用IL-1β的miRNA,结合芯片结果取交集。筛选5个交集中的miRNA和3个上调的miRNA并进行qRT-PCR验证。结果与假手术组比较,IS 2d组和IS 3d组大鼠TG中IL-1β升高。IS 3d组共有23个差异表达的miRNA(倍数>1.5,上调的有17个,下调的有6个)。qRT-PCR结果显示,1个上调的miRNA(miRNA-3560)及2个下调的miRNA(miRNA-760-3p、miRNA-130a-3p)与芯片结果一致(P<0.05)。结论 IS 3d组与假手术组大鼠TG中存在着差异表达的miRNA,miRNA-760-3p和miRNA-130a-3p可能调节IL-1β的表达,从而参与IL-1β在偏头痛中的分子机制。
Objective To screen microRNAs (miRNAs) that regulate IL-1β in the trigeminal ganglia (TG) of migraine rats. Methods Thirty-four SD rats were randomly divided into sham-operation group (n = 13), injection of 10μl ISI (IS 1d group, n = 4), 2 times (IS 2d group, n = 4) and 3 times (IS 3d group, n = 13). Western blot was used to detect the expression of IL-1β in TG. MiRNAs were extracted from RNA of rats in sham operation group and IS 3d group, and miRNAs of IL-1β target were predicted by software. Five miRNAs in the intersection and three up-regulated miRNAs were screened and verified by qRT-PCR. Results Compared with the sham-operation group, IL-1β in TG of IS 2d group and IS 3d group increased. There were 23 differentially expressed miRNAs in the IS 3d group (multiple> 1.5, up-regulated 17, down-regulated 6). qRT-PCR results showed that one miRNA (miRNA-3560) and two miRNAs (miRNA-760-3p, miRNA-130a-3p) were up-regulated (P <0.05). Conclusions There are differentially expressed miRNAs in TG of IS 3d group and sham operation group, miRNA-760-3p and miRNA-130a-3p may regulate the expression of IL-1β, and thus participate in the molecular mechanism of IL-1β in migraine .