目的:以人尿源性干细胞(human urine-derived stem cells,hUSCs)Wnt信号通路中的糖原合成酶激酶(glycogen synthase kinase-3β,GSK-3β)为靶标,从中药系统药理学(traditional Chinese medicine systems pharmacology,TCMSP)数据库收录...目的:以人尿源性干细胞(human urine-derived stem cells,hUSCs)Wnt信号通路中的糖原合成酶激酶(glycogen synthase kinase-3β,GSK-3β)为靶标,从中药系统药理学(traditional Chinese medicine systems pharmacology,TCMSP)数据库收录的中药小分子活性成分中筛选GSK-3β抑制剂,研究其是否具有增强hUSCs干性基因表达的作用。方法:参考京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)中人多能干细胞干性调控信号网络,以Wnt信号通路中的GSK-3β为靶点,利用分子对接技术对TCMSP数据库进行虚拟筛选,寻找能够增强hUSCs干性的中药小分子抑制剂;结合文献调研优化分子对接结果,通过MTT实验考察其对hUSCs细胞活力的影响,qRT-PCR实验考察其是否增强hUSCs干性基因的表达。结果:根据GSK-3β晶体结构复合的原配体对接结果,确定中药小分子活性成分的筛选规则为对接打分值高于阈值81.112(原配体打分值101.39的80%)且与受体GSK-3β的作用模式与原配体类似(与关键氨基酸残基ASP133和VAL135发生氢键作用);候选配体对接结果表明,黄芩素、白杨素、褪黑素与GSK-3β的对接打分值均高于阈值且与GSK-3β的作用模式均与原配体类似,可作为候选中药小分子活性成分;细胞实验结果表明,3种候选中药小分子活性成分在一定浓度范围内具有维持hUSCs细胞活力的作用,且均能显著上调hUSCs干性基因OCT4的表达水平,对MYC基因的表达水平无明显作用。结论:通过分子对接技术筛选到的针对GSK-3β的中药小分子抑制剂黄芩素、白杨素和褪黑素可以作为增强hUSCs干性的候选化合物,中药小分子活性成分有潜力被应用于hUSCs等人成体干细胞研究领域的药物研发。展开更多
In plants, each pollen mother cell undergoes two rounds of cell divisions to form a mature pollen grain, which contains a vegetative cell(VC) and two sperm cells(SC). As a companion cell, the VC carries the SCs to an ...In plants, each pollen mother cell undergoes two rounds of cell divisions to form a mature pollen grain, which contains a vegetative cell(VC) and two sperm cells(SC). As a companion cell, the VC carries the SCs to an ovule by germinating a pollen tube. In-depth sequencing analyses of mature pollen showed that micro RNAs(mi RNAs) and short interfering RNAs(si RNAs) are present in both the VC and SCs. Additionally, epigenetically-regulated transposable elements(TEs) are reactivated in the VC and these TE m RNAs are further processed into 21-nt epigenetically reactivated si RNA(easiR NA) in SCs, which prevent 24-nt si RNA accumulation and sequester mi RNA loading. Small RNAs are thought to move from the VC to SCs, where they regulate gene expression and reinforce TE silencing. Here, we summarize current knowledge of the biogenesis and function of mi RNAs, si RNAs, and easi RNAs in pollen, emphasizing how these different small RNAs coordinately contribute to sperm cell formation and TE silencing.展开更多
文摘目的:以人尿源性干细胞(human urine-derived stem cells,hUSCs)Wnt信号通路中的糖原合成酶激酶(glycogen synthase kinase-3β,GSK-3β)为靶标,从中药系统药理学(traditional Chinese medicine systems pharmacology,TCMSP)数据库收录的中药小分子活性成分中筛选GSK-3β抑制剂,研究其是否具有增强hUSCs干性基因表达的作用。方法:参考京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)中人多能干细胞干性调控信号网络,以Wnt信号通路中的GSK-3β为靶点,利用分子对接技术对TCMSP数据库进行虚拟筛选,寻找能够增强hUSCs干性的中药小分子抑制剂;结合文献调研优化分子对接结果,通过MTT实验考察其对hUSCs细胞活力的影响,qRT-PCR实验考察其是否增强hUSCs干性基因的表达。结果:根据GSK-3β晶体结构复合的原配体对接结果,确定中药小分子活性成分的筛选规则为对接打分值高于阈值81.112(原配体打分值101.39的80%)且与受体GSK-3β的作用模式与原配体类似(与关键氨基酸残基ASP133和VAL135发生氢键作用);候选配体对接结果表明,黄芩素、白杨素、褪黑素与GSK-3β的对接打分值均高于阈值且与GSK-3β的作用模式均与原配体类似,可作为候选中药小分子活性成分;细胞实验结果表明,3种候选中药小分子活性成分在一定浓度范围内具有维持hUSCs细胞活力的作用,且均能显著上调hUSCs干性基因OCT4的表达水平,对MYC基因的表达水平无明显作用。结论:通过分子对接技术筛选到的针对GSK-3β的中药小分子抑制剂黄芩素、白杨素和褪黑素可以作为增强hUSCs干性的候选化合物,中药小分子活性成分有潜力被应用于hUSCs等人成体干细胞研究领域的药物研发。
基金supported by grants from the National Natural Science Foundation of China(31422029,31470281)Shanghai Pujiang Program(13PJ1401200)to Zheng Bing Lian
文摘In plants, each pollen mother cell undergoes two rounds of cell divisions to form a mature pollen grain, which contains a vegetative cell(VC) and two sperm cells(SC). As a companion cell, the VC carries the SCs to an ovule by germinating a pollen tube. In-depth sequencing analyses of mature pollen showed that micro RNAs(mi RNAs) and short interfering RNAs(si RNAs) are present in both the VC and SCs. Additionally, epigenetically-regulated transposable elements(TEs) are reactivated in the VC and these TE m RNAs are further processed into 21-nt epigenetically reactivated si RNA(easiR NA) in SCs, which prevent 24-nt si RNA accumulation and sequester mi RNA loading. Small RNAs are thought to move from the VC to SCs, where they regulate gene expression and reinforce TE silencing. Here, we summarize current knowledge of the biogenesis and function of mi RNAs, si RNAs, and easi RNAs in pollen, emphasizing how these different small RNAs coordinately contribute to sperm cell formation and TE silencing.