miR-153-3p调控神经突的生长

Abstract

目的探讨miR-153-3p在大、小鼠神经突生长中的作用。方法对数生长期C57BL/6小鼠N2A细胞和Rattus norvegicus大鼠PC12细胞,随机分为miR-153-3p过表达组(转染miR-153-3p过表达试剂miR-153-3p mimic)、miR-153-3p抑制组(转染miR-153-3p表达抑制剂miR-153-3p inhibitor)、过表达对照组(转染miR-153-3p mimic空白对照试剂miR-153-3p mimic NC)、抑制剂对照组(转染miR-153-3p inhibitor空白对照试剂miR-153-3p inhibitor NC)。转染48 h,采用实时荧光定量PCR法检测4组细胞miR-153-3p mRNA相对表达量,免疫荧光法检测神经突生长情况。结果 miR-153-3p过表达组N2A细胞、PC12细胞miR-153-3p mRNA相对表达量(28.98±1.63、19.67±0.88)均高于miR-153-3p抑制组(0.94±0.19、0.89±0.05)、过表达对照组(0.99±0.08、1.00±0.09)、抑制剂对照组(1.00±0.04、1.00±0.14)(P<0.05),miR-153-3p抑制组、过表达对照组、抑制剂对照组比较差异均无统计学意义(P>0.05);miR-153-3p过表达组N2A细胞、PC12细胞神经突总长度[(31.13±3.85)、(38.08±2.12)μm]和最长神经轴突长度[(18.04±2.58)、17.48±1.75)μm]均较miR-153-3p抑制组[N2A细胞:(54.53±1.83)、(31.74±2.36)μm;PC12细胞:(64.46±12.85)、(29.46±7.36)μm]、过表达对照组[N2A细胞:(42.53±6.29)、(24.99±3.47)μm;PC12细胞:(53.58±4.88)、(25.31±3.66)μm]、抑制剂对照组[N2A细胞:(41.33±2.77)、(24.57±3.86)μm;PC12细胞:(44.44±7.59)、(23.04±4.31)μm]短(P<0.05),且过表达对照组、抑制剂对照组PC12细胞、N2A细胞神经突总长度和最长神经轴突长度较miR-153-3p抑制组短(P<0.05),过表达对照组、抑制剂对照组比较差异无统计学意义(P>0.05)。结论miR-153-3p可抑制小鼠N2A细胞、大鼠PC12细胞神经突生长。

Objective To explore the role of miR-153-3 p in regulating neurite outgrowth in mice and rats. Methods The N2 A cells from C57 BL/6 mice and PC12 cells from Rattus norvegicus rats in logarithmic growth phase were randomly divided into four groups: overexpression group transfected with miR-153-3 p mimic, inhibition group transfection with miR-153-3 p inhibitor, overexpression control group transfected with miR-153-3 p mimic NC and inhibition control group transfected with miR-153-3 p inhibitor NC. The relative expression of miR-153-3 p was detected by real-time fluorescence quantitative PCR, and the neurite outgrowth was observed by immunofluorescence assay in four groups after 48 h transfection. Results The relative expression levels of miR-153-3 p mRNA in N2 A cells and PC12 cells were significantly higher in overexpression group(28.98±1.63, 19.67±0.88) than those in inhibition group(0.94±0.19, 0.89±0.05), overexpression control group(0.99±0.08, 1.00±0.09) and inhibition control group(1.00±0.04, 1.00±0.14)(P<0.05), and showed no significant differences among inhibition group, overexpression control group and inhibition control group(P>0.05). The total neurite lengths and the longest neurite lengths of N2 A cells and PC12 cells were significantly shorter in overexpression group(N2 A:(31.13±3.85),(18.04±2.58) μm; PC12:(38.08±2.12),(17.48±1.75) μm) than those in inhibition group(N2 A:(54.53±1.83),(31.74±2.36) μm; PC12:(64.46±12.85),(29.46±7.36) μm), overexpression control group(N2 A:(42.53±6.29),(24.99±3.47) μm; PC12:(53.58±4.88),(25.31±3.66) μm) and inhibition control group(N2 A:(41.33±2.77),(24.57±3.86) μm; PC12:(44.44±7.59),(23.04±4.31) μm)(P<0.05), shorter in overexpression control group and inhibition control group than those in inhibition group(P<0.05), and showed no significant differences between overexpression control group and inhibition control group(P>0.05). Conclusion MiR-153-3 p could inhibit the neurite outgrowth of N2 A cells in mice and PC12 cells in rats.