右美托咪定通过miR-142-3p/昼夜节律基因通路调节神经元增殖及凋亡研究

Dexmedetomidine regulates neuron cell proliferation and apoptosis through miR-142-3p/circadian rhythm gene

目的本研究旨在探讨右美托咪定通过miR-142-3p作用于昼夜节律基因(ARNTL)调节神经元增殖及凋亡的影响。方法将一半大鼠海马元代神经元细胞分为对照组(未进行任何处理)和实验组(50μmol·L^-1右美托咪定),对照组予以等体积的培养基液。其次,再将另一半细胞大鼠海马元代神经元细胞分为空白组(未进行任何处理)、过表达组(转染miR-142-3p模拟物)、抑制表达组(转染miR-142-3p抑制剂;通过实时荧光定量聚合酶链反应和蛋白质印记法检测miR-142-3p及ARNTL表达;通过四甲基偶氮唑盐微量酶反应比色法(MTT)检测细胞增殖;通过流式细胞术检测细胞凋亡。通过双荧光素酶报告基因实验验证ARNTL与miR-142-3p的靶向关系。结果给药48 h后,实验组和对照组的海马神经元细胞增殖率分别为(1.65±0.08)%和(1.03±0.06)%;细胞凋亡率分别为(3.94±0.15)%和(9.65±0.13)%;miR-142-3p相对表达量分别为1.66±0.14和1.05±0.12;ARNTL基因相对表达量分别为0.85±0.06和0.96±0.19,差异均有统计学意义(均P<0.05)。空白组、过表达组和抑制表达组的miR-142-3p基因相对表达量分别为1.06±0.11,5.24±0.32和0.52±0.06;ARNTL基因相对表达分别为1.02±0.09,0.63±0.12和3.46±0.14;细胞增殖率分别为(1.11±0.10)%,(1.72±0.09)%和(0.62±0.08)%;细胞凋亡率分别为(5.44±0.16)%,(3.56±0.15)%和(7.52±0.15)%,过表达组分别与空白组和抑制表达组比较,差异均有统计学意义(均P<0.05)。结论右美托咪定通过miR-142-3p/ARNTL增强海马神经元细胞增殖、抑制神经元细胞凋亡。

Objective The aim of this study was to investigate the effect of dexmedetomidine on neuron cell proliferation and apoptosis through miR-142-3 p/circadian rhythm gene(ARNTL). Methods Half number of primary hippocampal neuron cells were assigned to the control group and the test group(50 μmol·L^-1 dexmedetomidine), and then primary hippocampal neuron cells were divided into blank group(without any treatment), overexpression group(transfected with miR-142-3 p mimic) and inhibitory expression group(transfected with miR-142-3 p inhibitor), the expression of miR-142-3 p and ARNTL was detected by real-time fluorescent quantitative polymerase chain reaction and protein imprinting, cell proliferation was detected by enzyme reaction colorimetry of tetramethylazo salt(MTT) assay,apoptosis was detected by flow cytometry. The targeting relationship between ARNTL and miR-142-3 p was verified by double luciferase reporter gene experiment. Results At 48 h after treatment,there were statistically significant differences between the test group and the control group in the proliferation rate of hippocampal neuron cells [(1. 65 ± 0. 08) % vs(1. 03 ± 0. 06) % ], the apoptosis rates[(3. 94 ± 0. 15) % vs(9. 65 ± 0. 13) % ],the relative expression of miR-142-3 p [(1. 66 ± 0. 14) vs(1. 05 ± 0. 12) ],the relative expression of ARNTL gene [(0. 85 ± 0. 06) vs(0. 96 ± 0. 19) ](all P < 0. 05). The relative expression levels of miR-142-3 p in the blank group,overexpression group and the inhibition group were1. 06 ± 0. 11,5. 24 ± 0. 32 and 0. 52 ± 0. 06;the relative expression levels of ARNTL were 1. 02 ± 0. 09,0. 63 ± 0. 12 and 3. 46 ± 0. 14;the cell proliferation rates were(1. 11 ± 0. 10) %,(1. 72 ± 0. 09) % and(0. 62 ± 0. 08) %;the apoptosis rates were(5. 44 ± 0. 16) %,(3. 56 ± 0. 15) %,and(7. 52 ± 0. 15) %. There were statistically significant differences between the overexpression group and the blank group/inhibition group(all P < 0. 05). Conclusion Dexmedetomidine can enhance the proliferation and inhibit the apoptosis of hippocampal neuron cells through miR-142-3 p/ARNTL.