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 the microRNAs(miRNA)in trigeminal ganglia(TG)regulating IL-1βin rat model with migraine.Methods Parasagittal epidural catheterization was performed in 34 SD rats,which were randomly divided into four groups of C(sham-operated,n=13),IS1(infusion of inflammation soup 10μl/d for one day,n=4),IS2(for 2days,n=4)and IS3(for3days,n=13).Western blot analysis was used to detect the expression of IL-1βin TG.The RNA in rat TG in groups of C and IS3 was extracted for detecting aberrant miRNA regulation using the miRNA microarray.Software was applied to predict miRNAs that directly targeted at IL-1β.Then the predicted miRNAs belonging to down-regulated miRNAs were validated via qRT-PCR.Results Compared with group C,IL-1βin TG was increased in groups of IS2 and IS3.Of 23 aberrant miRNAs in group IS3,the expressions of 17 miRNAs were up-regulated and 6miRNAs were down-regulated.The results of qRT-PCR were consistent with those of microarray,which showed that miRNA-3560 was up-regulated and two miRNAs of miRNA-760-3p and miRNA-130a-3p were down-regulated(P〈0.05).Conclusion The differential expression of miRNA in TG exists between groups of IS3 and C.miRNA-760-3p and miRNA-130a-3p may participate in the molecular mechanisms of migraine by regulating the expression of IL-1β.