摘要
目的研究microRNA-26b对糖氧剥夺(oxygen-glucose deprivation,OGD)诱导BV-2细胞释放白介素-6(interleukin6,IL-6)的影响。方法 OGD诱导小鼠胶质细胞系BV-2细胞活化,通过ELISA和RT-qPCR定量培养上清IL-6浓度及BV-2细胞IL-6 mRNA水平。转染IL-6microRNA-26b模拟物(miR-26b mimics)和阴性对照(negative control,NC)至BV-2细胞后,经OGD处理检测培养上清IL-6浓度。通过共转miR-26b mimics和携带有IL-6 mRNA3'UTR的双荧光素酶报告质粒至293T细胞,明确miR-26b对IL-6的靶向调节作用。结果与对照组比较,经1 h OGD处理后的BV-2细胞IL-6 mRNA水平和培养上清中IL-6增加(P<0.05);转染microRNA-26b可以减少OGD引起的IL-6释放(P<0.05);共转miR-26b和双荧光素酶报告质粒后萤火虫荧光素酶活性高于对照组(P<0.05)。结论microRNA-26b可结合至IL-6 mRNA的3'UTR区,可能通过这一机制下调OGD诱导的BV-2细胞IL-6表达。
Objective To investigate the effects of miR-26b on OGD-induced IL-6 expression of BV-2 cells. Methods BV-2 cells were processed with OGD. The level of IL-6 in culture medium was tested by enzyme-linked immunosorbent assay （ELISA）. The quantitative real-time PCR was used to measure the expression of IL-6 mRNA in BV-2 cells. BV-2 cells transfected with microRNA-26b mimics were processed with OGD and tested IL-6 level of the culture medium. The miR-26b targeting IL-6 was further determined by co-transfecting of miR-26b mimics and dual luciferase reporter vector to 293T cells. Results IL-6 mRNA and protein were up-regulated in OGD-treated BV2 cells （P〈0.05）. IL-6 in OGD-treated BV-2 decreased after transfected with miR-26b mimics （P〈0.05）. Dual luciferase reporter assay suggested the binding of miR-26b to the 3＇UTR of mouse IL-6 mRNA. Conclusions MicroRNA-26b can combine to 3＇UTR of IL-6 mRNA and may negatively regulate the expression of IL-6 through this mechanism in OGD-induced BV-2 cells.
出处
《老年医学与保健》
CAS
2016年第2期85-88,共4页
Geriatrics & Health Care
基金
国家自然科学基金(81371220)
