Spinal cord injury typically causes corticospinal tract disruption.Although the disrupted corticospinal tract can self-regenerate to a certain degree,the underlying mechanism of this process is still unclear.N6-methyl...Spinal cord injury typically causes corticospinal tract disruption.Although the disrupted corticospinal tract can self-regenerate to a certain degree,the underlying mechanism of this process is still unclear.N6-methyladenosine(m^(6)A)modifications are the most common form of epigenetic regulation at the RNA level and play an essential role in biological processes.However,whether m^(6)A modifications participate in corticospinal tract regeneration after spinal cord injury remains unknown.We found that expression of methyltransferase 14 protein(METTL14)in the locomotor cortex was high after spinal cord injury and accompanied by elevated m^(6)A levels.Knockdown of Mettl14 in the locomotor cortex was not favorable for corticospinal tract regeneration and neurological recovery after spinal cord injury.Through bioinformatics analysis and methylated RNA immunoprecipitation-quantitative polymerase chain reaction,we found that METTL14 regulated Trib2 expression in an m^(6)A-regulated manner,thereby activating the mitogen-activated protein kinase pathway and promoting corticospinal tract regeneration.Finally,we administered syringin,a stabilizer of METTL14,using molecular docking.Results confirmed that syringin can promote corticospinal tract regeneration and facilitate neurological recovery by stabilizing METTL14.Findings from this study reveal that m^(6)A modification is involved in the regulation of corticospinal tract regeneration after spinal cord injury.展开更多
[目的]探究Lnc-AC226118.1/miR-99b-5p/TRIB1信号通路对术后认知障碍大鼠神经细胞活性的影响。[方法]将术后认知障碍大鼠神经细胞分为3组:对照组、si NC组和si TRIB1组。通过CCK-8实验检测神经细胞增殖能力;通过细胞集落形成实验检测神...[目的]探究Lnc-AC226118.1/miR-99b-5p/TRIB1信号通路对术后认知障碍大鼠神经细胞活性的影响。[方法]将术后认知障碍大鼠神经细胞分为3组:对照组、si NC组和si TRIB1组。通过CCK-8实验检测神经细胞增殖能力;通过细胞集落形成实验检测神经细胞聚集活性;通过流式细胞数检测神经细胞凋亡率;通过生物信息学分析Lnc-AC226118.1/miR-99b-5p/TRIB1信号轴的结合位点;通过荧光素报告基因实验检测Lnc-AC226118.1/miR-99b-5p/TRIB1信号轴的靶向作用关系。通过蛋白免疫印迹实验分析TRIB1的表达。通过实时荧光PCR实验检测miR-99b-5p的表达。[结果]与对照组、si NC组相比,si TRIB1组的神经细胞增殖和聚集活性增加(1.25±0.08 vs 1.36±0.11 vs 2.27±0.03;62.69±12.28 vs 57.36±13.59 vs 173.51±22.68;P<0.05),凋亡率降低(13.19%±0.29%vs 14.66%±0.52%vs 3.69%±0.08%;P<0.05)。Lnc-AC226118.1/miR-99b-5p/TRIB1信号轴分子间有相互结合位点,且具有靶向调控关系。[结论]激活Lnc-AC226118.1/miR-99b-5p/TRIB1信号轴能够提高术后认知障碍大鼠神经细胞活性,Lnc-AC226118.1/miR-99b-5p/TRIB1信号轴有望成为术后认知障碍治疗的潜在靶点。展开更多
基金supported by the National Natural Science Foundation of China,Nos.82030071(to JH),82272495(to YC)Science and Technology Major Project of Changsha,No.kh2103008(to JH)Graduate Students’Independent Innovative Projects of Hunan Province,No.CX20230311(to YJ)。
文摘Spinal cord injury typically causes corticospinal tract disruption.Although the disrupted corticospinal tract can self-regenerate to a certain degree,the underlying mechanism of this process is still unclear.N6-methyladenosine(m^(6)A)modifications are the most common form of epigenetic regulation at the RNA level and play an essential role in biological processes.However,whether m^(6)A modifications participate in corticospinal tract regeneration after spinal cord injury remains unknown.We found that expression of methyltransferase 14 protein(METTL14)in the locomotor cortex was high after spinal cord injury and accompanied by elevated m^(6)A levels.Knockdown of Mettl14 in the locomotor cortex was not favorable for corticospinal tract regeneration and neurological recovery after spinal cord injury.Through bioinformatics analysis and methylated RNA immunoprecipitation-quantitative polymerase chain reaction,we found that METTL14 regulated Trib2 expression in an m^(6)A-regulated manner,thereby activating the mitogen-activated protein kinase pathway and promoting corticospinal tract regeneration.Finally,we administered syringin,a stabilizer of METTL14,using molecular docking.Results confirmed that syringin can promote corticospinal tract regeneration and facilitate neurological recovery by stabilizing METTL14.Findings from this study reveal that m^(6)A modification is involved in the regulation of corticospinal tract regeneration after spinal cord injury.
文摘[目的]探究Lnc-AC226118.1/miR-99b-5p/TRIB1信号通路对术后认知障碍大鼠神经细胞活性的影响。[方法]将术后认知障碍大鼠神经细胞分为3组:对照组、si NC组和si TRIB1组。通过CCK-8实验检测神经细胞增殖能力;通过细胞集落形成实验检测神经细胞聚集活性;通过流式细胞数检测神经细胞凋亡率;通过生物信息学分析Lnc-AC226118.1/miR-99b-5p/TRIB1信号轴的结合位点;通过荧光素报告基因实验检测Lnc-AC226118.1/miR-99b-5p/TRIB1信号轴的靶向作用关系。通过蛋白免疫印迹实验分析TRIB1的表达。通过实时荧光PCR实验检测miR-99b-5p的表达。[结果]与对照组、si NC组相比,si TRIB1组的神经细胞增殖和聚集活性增加(1.25±0.08 vs 1.36±0.11 vs 2.27±0.03;62.69±12.28 vs 57.36±13.59 vs 173.51±22.68;P<0.05),凋亡率降低(13.19%±0.29%vs 14.66%±0.52%vs 3.69%±0.08%;P<0.05)。Lnc-AC226118.1/miR-99b-5p/TRIB1信号轴分子间有相互结合位点,且具有靶向调控关系。[结论]激活Lnc-AC226118.1/miR-99b-5p/TRIB1信号轴能够提高术后认知障碍大鼠神经细胞活性,Lnc-AC226118.1/miR-99b-5p/TRIB1信号轴有望成为术后认知障碍治疗的潜在靶点。
文摘采用PCR直接测序法对伞形科（ Apiaceae）前胡族（ Trib. Peucedaneae）种类川明参（ Chuanminshen violaceum Sheh et Shan）、泰山前胡〔Peucedanum wawrae （ H. Wolff） Su〕和华中前胡（ P. medicum Dunn）以及美味芹族（ Trib. Smyrnieae）种类明党参（Changium smyrnioides H. Wolff）、宝兴棱子芹（Pleurospermum davidii Franch.）、丽江棱子芹（ P. foetens Franch.）和鸡冠棱子芹（ P. cristatum de Boiss.）的叶绿体基因组psbA-trnH片段进行了扩增测序,获得的序列已提交至GenBank,登录号为KF557756-KF557762。结合引自GenBank的前胡族阿魏属（ Ferula Linn.）1种、大瓣芹属（ Semenovia Regel et Herder）1种、当归属（ Angelica Linn.）2种和美味芹族的舟瓣芹属（ Sinolimprichtia H. Wolff）1种、羌活属（ Notopterygium de Boiss.）1种、瘤果芹属（ Trachydium Lindl.）1种以及针果芹族（ Trib. Scandicineae）刺果芹〔Turgenia latifolia （ Linn.） Hoffm.〕的psbA-trnH片段序列,对各种类的psbA-trnH片段信息进行分析；并以刺果芹为外类群构建了MP、ML和BI系统发育树。结果表明：川明参和明党参的psbA-trnH片段长度均为258 bp、GC含量均为23％,而其他种类的psbA-trnH片段长度为228~405 bp、GC含量为26％~35％；排序后psbA-trnH 序列总长度为553 bp（包括空位）,其中变异位点237个、信息位点178个。川明参与明党参间的相对遗传距离最小（仅为0.02）,而川明参与其他种类间的相对遗传距离为0.10~1.34,且总体上川明参与美味芹族种类的相对遗传距离较小,表明川明参与明党参及美味芹族种类的亲缘关系较近。在3类系统树上,川明参与明党参均聚在一起,并与美味芹族的属种聚为一大支,而远离由前胡族属种构成的另一大支。结合外部形态和果实解剖结构特征,建议将川明参属（ Chuanminshen Sheh et Shan ）从前胡族中分出并置于美味芹族中,与明党参属（ Changium H. Wolff）为姐妹类群。
