摘要
该研究筛选了能特异性结合和稳定结核分枝杆菌Rv0341基因G4结构的小分子化合物。采用圆二色谱法探究Rv0341上富G寡核苷酸链能否形成G4(Rv0341G4)结构,并采用实时荧光定量反转录聚合酶链反应(qRT-PCR)和蛋白质印迹法验证小分子化合物对Rv0341表达的抑制作用。通过表面等离子体共振(SPR)技术筛选得与Rv0341G4结合的小分子化合物T-007;qRT-PCR法和蛋白质印迹法分析显示T-007可稳定Rv0341G4结构,对Rv0341转录具有负调控作用。该研究可为靶向GR结构的结核病治疗药物开发提供一定参考。
This study screened small molecule compounds capable of specifically binding to and stabilizing the G-quadruplex(G4)structure within the Rv0341 gene of Mycobacterium tuberculosis.Firstly,circular dichroism(CD)spectroscopy was used to confirm the formation of the G4 structure(designated Rv0341G4)by the G-rich oligonucleotide sequence in Rv0341 gene.Subsequently,real time fluorescent quantitative reverse transcription polymerase chain reaction(qRT-PCR)and Western blot were employed to validate the inhibitory effects of candidate small molecules on Rv0341 expression.Through surface plasmon resonance(SPR)screening,a small molecule compound T-007 exhibiting high-affinity binding to Rv0341G4 was identified.Both qRT-PCR and Western blot analyses demonstrated that T-007 stabilizes the Rv0341G4 structure and downregulates Rv0341 transcription.This study provides valuable insights for developing novel anti-tuberculosis therapeutics targeting G4 structures.
作者
王昱斌
都甜甜
颉亚辉
潘云燕
WANG Yubin;DU Tiantian;XIE Yahui;PAN Yunyan(Laboratory Medicine Center of the Second Hospital&Clinical Medical School,Lanzhou University,Lanzhou 730030;Nephrology Department of the Second Hospital&Clinical Medical School,Lanzhou University,Lanzhou 730030;School of Public Health,Gansu University of Traditional Chinese Medicine,Lanzhou 730030)
出处
《中国医药工业杂志》
2025年第7期886-892,共7页
Chinese Journal of Pharmaceuticals
基金
甘肃省青年科技基金项目(21JR11RA115)
甘肃省教育厅2023高校教师创新基金项目(2023B-104)
临床萃英拔尖项目(CY2023-RJ-08)。
关键词
结核分枝杆菌
圆二色谱
Rv0341基因G4结构
表面等离子体共振
实时荧光定量反转录PCR
蛋白质印迹
抗结核药
Mycobacterium tuberculosis
circular dichroism spectroscopy
G-quadruplex structure within the Rv0341 gene
surface plasmon resonance
real time fluorescent quantitative reverse transcription polymerase chain reaction
Western blot
antituberculotic drug
