CircRNA regulates microglia polarization in diabetic retinopathy

CircRNA expression profiles of mice microglia cells BV2 treated with or without glucose were detected. Differentially expressed circRNAs were confirmed by qRT-PCR and bioinformatics analysis was performed. The expression of the key circRNA, mm9_circ_01468, as well as its biological characteristics, was determined by PCR assay. The siRNA for mm9_circ_01468 and the mm9_circ_01468 overexpressing lentiviruses were constructed to investigate its effect in the proliferation, apoptosis and polarization of microglia. RNA sequencing was performed to identify the key genes and signaling pathways in mm9_circ_01468 knocking down BV2. Lentiviriruses encoding shRNA for mm9_circ_01468 (Lv-circRNA) were constructed for intraocular injection.

本研究检测了经葡萄糖处理与未处理的小鼠小胶质细胞BV2的环状RNA（circRNA）表达谱。通过实时定量聚合酶链反应（qRT-PCR）验证了差异表达的环状RNA，并开展了生物信息学分析。通过聚合酶链反应（PCR）实验测定了关键环状RNA mm9_circ_01468的表达水平及其生物学特性。本研究构建了针对mm9_circ_01468的小干扰RNA（siRNA）与mm9_circ_01468过表达慢病毒，以探究该环状RNA对小胶质细胞增殖、凋亡及极化的调控作用。通过RNA测序（RNA-seq）技术，鉴定了mm9_circ_01468敲低的BV2细胞中的关键基因与信号通路。本研究构建了携带mm9_circ_01468短发夹RNA（shRNA）的慢病毒（Lv-circRNA），用于眼内注射实验。