Somatic cells respond to considerable stress,and go through a series of phytohormone pathways,then forming an embryo.The developmental process is recorded as somatic embryogenesis(SE).One of the key components regulat...Somatic cells respond to considerable stress,and go through a series of phytohormone pathways,then forming an embryo.The developmental process is recorded as somatic embryogenesis(SE).One of the key components regulating SE are the microRNAs(miRNAs).Despite previous studies,it is still not clear exactly how miRNAs exert their function of regulating targets during conditionally activated early SE.Here,we use Liriodendron sino-americanum as a model system and perform a combined analysis of microfluidic chips and degradome sequencing to study this process.We identified a total of 386 conserved miRNAs and 153 novel miRNAs during early SE.According to the ANOVA test,239 miRNAs showed 12 distinct expression patterns.Through degradome sequencing,419 targets and 198 targets were identified for 136 known miRNAs and 37 novel miRNAs,respectively.Gene Ontology(GO)and metabolism pathway enrichment analysis revealed that these targets were significantly involved in oxidation-reduction processes,calmodulin-mediated signal transduction pathways and carbohydrate metabolism.The genes that were related to stress responses,phytohormone pathways and plant metabolism were identified within the targets of miR319,miR395,miR408,miR472,miR482,miR390,miR2055,miR156,miR157,miR171,miR396,miR397,miR529,miR535 and miR159.According to promoter analysis,various cis-acting elements related to plant growth and development,phytohormones response and stress response were present in the promoter of the miRNAs.The differential expression patterns of 11 miRNA-target modules were confirmed by real-time quantitative PCR.The study demonstrated that the miRNA plays an important role in the early SE process by regulating its target and then participating in carbohydrate metabolism and stress response.It also provided a valuable resource for further research in determining the genetic mechanism of SE,and then facilitating breeding programs on plants.展开更多
目的:评估芍药苷通过信号转导与转录激活子3(signal transducer and activator of transcriptions 3,STAT3)对白细胞介素-13(interleukin-13,IL-13)诱导的BEAS-2B哮喘细胞模型的炎症、氧化应激和自噬的改善作用。方法:用1、10和30μmol&...目的:评估芍药苷通过信号转导与转录激活子3(signal transducer and activator of transcriptions 3,STAT3)对白细胞介素-13(interleukin-13,IL-13)诱导的BEAS-2B哮喘细胞模型的炎症、氧化应激和自噬的改善作用。方法:用1、10和30μmol·L^(-1)的芍药苷干预IL-13诱导前后的BEAS-2B细胞,CCK-8检测细胞增殖活力,筛选芍药苷最佳作用条件。然后利用芍药苷和STAT3激活剂干预IL-13诱导的细胞模型,将细胞分为对照组、模型组、STAT3激活剂组、芍药苷组和STAT3激活剂+芍药苷共处理组。CCK-8检测各组细胞增殖活力,ELISA试剂盒检测各组细胞上清中IL-4、IFN-γ的含量,及细胞中MDA、CAT、SOD水平,Western blot检测各组细胞LC3Ⅱ、LC3Ⅰ、P62、p-STAT3和STAT3的蛋白表达。结果:CCK-8检测结果显示,10μmol·L^(-1)的芍药苷作用24 h为最佳干预条件。与对照组相比,模型组细胞增殖能力下降(P<0.05),IFN-γ、SOD和CAT含量下降(P<0.05),IL-4和MDA含量增加(P<0.05),LC3Ⅱ/Ⅰ蛋白表达比值和p-STAT3/STAT3蛋白表达比值增加(P<0.05),P62蛋白表达下降(P<0.05)。与模型组相比,STAT3激动剂组细胞增殖能力下降(P<0.05),IFN-γ、SOD和CAT含量下降(P<0.05),IL-4和MDA含量增加(P<0.05),LC3Ⅱ/Ⅰ蛋白表达比值和p-STAT3/STAT3蛋白表达比值增加(P<0.05),P62蛋白表达下降(P<0.05);芍药苷组细胞增殖能力增加(P<0.05),IFN-γ、SOD和CAT含量增加(P<0.05),IL-4和MDA含量下降(P<0.05),LC3Ⅱ/Ⅰ蛋白表达比值和p-STAT3/STAT3蛋白表达比值下降(P<0.05),P62蛋白表达增加(P<0.05)。与STAT3激动剂组相比,STAT3激动剂+芍药苷组共处理逆转了STAT3激动剂的作用。结论:芍药苷通过抑制STAT3蛋白磷酸化,抑制炎症反应,从而改善哮喘中细胞氧化应激和自噬。展开更多
基金supported by the Natural Science Foundation of China[32071784]the Qinglan Project of Jiangsu Provincethe Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD).
文摘Somatic cells respond to considerable stress,and go through a series of phytohormone pathways,then forming an embryo.The developmental process is recorded as somatic embryogenesis(SE).One of the key components regulating SE are the microRNAs(miRNAs).Despite previous studies,it is still not clear exactly how miRNAs exert their function of regulating targets during conditionally activated early SE.Here,we use Liriodendron sino-americanum as a model system and perform a combined analysis of microfluidic chips and degradome sequencing to study this process.We identified a total of 386 conserved miRNAs and 153 novel miRNAs during early SE.According to the ANOVA test,239 miRNAs showed 12 distinct expression patterns.Through degradome sequencing,419 targets and 198 targets were identified for 136 known miRNAs and 37 novel miRNAs,respectively.Gene Ontology(GO)and metabolism pathway enrichment analysis revealed that these targets were significantly involved in oxidation-reduction processes,calmodulin-mediated signal transduction pathways and carbohydrate metabolism.The genes that were related to stress responses,phytohormone pathways and plant metabolism were identified within the targets of miR319,miR395,miR408,miR472,miR482,miR390,miR2055,miR156,miR157,miR171,miR396,miR397,miR529,miR535 and miR159.According to promoter analysis,various cis-acting elements related to plant growth and development,phytohormones response and stress response were present in the promoter of the miRNAs.The differential expression patterns of 11 miRNA-target modules were confirmed by real-time quantitative PCR.The study demonstrated that the miRNA plays an important role in the early SE process by regulating its target and then participating in carbohydrate metabolism and stress response.It also provided a valuable resource for further research in determining the genetic mechanism of SE,and then facilitating breeding programs on plants.
文摘目的:评估芍药苷通过信号转导与转录激活子3(signal transducer and activator of transcriptions 3,STAT3)对白细胞介素-13(interleukin-13,IL-13)诱导的BEAS-2B哮喘细胞模型的炎症、氧化应激和自噬的改善作用。方法:用1、10和30μmol·L^(-1)的芍药苷干预IL-13诱导前后的BEAS-2B细胞,CCK-8检测细胞增殖活力,筛选芍药苷最佳作用条件。然后利用芍药苷和STAT3激活剂干预IL-13诱导的细胞模型,将细胞分为对照组、模型组、STAT3激活剂组、芍药苷组和STAT3激活剂+芍药苷共处理组。CCK-8检测各组细胞增殖活力,ELISA试剂盒检测各组细胞上清中IL-4、IFN-γ的含量,及细胞中MDA、CAT、SOD水平,Western blot检测各组细胞LC3Ⅱ、LC3Ⅰ、P62、p-STAT3和STAT3的蛋白表达。结果:CCK-8检测结果显示,10μmol·L^(-1)的芍药苷作用24 h为最佳干预条件。与对照组相比,模型组细胞增殖能力下降(P<0.05),IFN-γ、SOD和CAT含量下降(P<0.05),IL-4和MDA含量增加(P<0.05),LC3Ⅱ/Ⅰ蛋白表达比值和p-STAT3/STAT3蛋白表达比值增加(P<0.05),P62蛋白表达下降(P<0.05)。与模型组相比,STAT3激动剂组细胞增殖能力下降(P<0.05),IFN-γ、SOD和CAT含量下降(P<0.05),IL-4和MDA含量增加(P<0.05),LC3Ⅱ/Ⅰ蛋白表达比值和p-STAT3/STAT3蛋白表达比值增加(P<0.05),P62蛋白表达下降(P<0.05);芍药苷组细胞增殖能力增加(P<0.05),IFN-γ、SOD和CAT含量增加(P<0.05),IL-4和MDA含量下降(P<0.05),LC3Ⅱ/Ⅰ蛋白表达比值和p-STAT3/STAT3蛋白表达比值下降(P<0.05),P62蛋白表达增加(P<0.05)。与STAT3激动剂组相比,STAT3激动剂+芍药苷组共处理逆转了STAT3激动剂的作用。结论:芍药苷通过抑制STAT3蛋白磷酸化,抑制炎症反应,从而改善哮喘中细胞氧化应激和自噬。
