胆红素对神经干细胞活性和增殖的影响研究
Effect of bilirubin on the viability and proliferation of neural stem cells
吴翠萍 1朱一丹 1李春燕 1殷善开1
作者信息
- 1. 上海交通大学医学院附属第六人民医院耳鼻咽喉头颈外科/上海市睡眠呼吸障碍疾病重点实验室/上海交通大学耳鼻咽喉研究所,上海 200233
- 折叠
摘要
目的 探究胆红素对神经干细胞(neural stem cells,NSCs)活性和增殖的影响.方法 分离培养小鼠NSCs,加入不同浓度的胆红素,培养 2d后行活/死细胞染色;培养 7d后行神经球形成、Ki-67 检测和转录组测序分析以探究胆红素对NSCs活性和增殖的影响.结果 1.71～6.84 μmol/L 胆红素对 NSCs 的活性无影响,13.68 μmol/L 胆红素降低了 NSCs 的活性.0.17～6.84 μmol/L胆红素促进NSCs的增殖,当胆红素的浓度达到 13.68 μmol/L时,促进增殖的作用消失.胆红素促进了NSCs的Ki-67 表达.转录组结果显示胆红素组 184 个基因上调表达和 100 个基因下调表达,差异基因与"细胞群体增殖调节"等生物学过程和PI3K-AKT等信号通路相关.结论 高浓度的胆红素对NSCs具有毒性作用,低浓度的胆红素不影响NSCs活性,并促进NSCs增殖.
Abstract
Objective To investigate the effect and mechanism of bilirubin on neural stem cell(NSC)viability and proliferation.Methods Primary mouse NSCs were cultured with different concentrations of bilirubin,and live-dead cell staining was performed after 2 days of culture;neurosphere assay,Ki-67 assay,and transcriptome sequencing were performed after 7 days of culture to in-vestigate the effects of bilirubin on NSC viability and proliferation.Results Approximately 1.71-6.84 μmol/L bilirubin had no effect on NSC viability,while 13.68 μmol/L bilirubin decreased NSC viability.Approximately 0.17-6.84 μmol/L bilirubin promo-ted NSC proliferation,while this pro-proliferation effect disappeared when the bilirubin concentration reached 13.68 μmol/L.Biliru-bin increased Ki-67 expression in NSCs.RNA Seq showed that bilirubin upregulated 184 genes and downregulated 100 genes.Gene Ontology analysis suggested that the differentially expressed genes were mainly related to"regulation of cell population proliferation"and other biological processes.KEGG analysis suggested that the differentially expressed genes were related to several pathways,such as the"PI3K-AKT signaling pathway".Conclusion High bilirubin concentrations were toxic to NSCs,while low bilirubin concentrations had no effect on NSC viability and promoted NSC proliferation.
关键词
胆红素/神经干细胞/增殖/转录组测序Key words
Bilirubin/Neural stem cells/Proliferation/RNA-Seq引用本文复制引用
基金项目
国家自然科学基金(82071042)
上海市自然科学基金(20ZR1442300)
出版年
2024
NETL
NSTL
万方数据