中国药理学与毒理学杂志2025，Vol.39Issue(7)：489-499,11.DOI:10.3867/j.issn.1000-3002.2025.08393
脑缺血再灌注损伤对脑内IMM-H004葡萄糖醛酸化代谢的影响
Effect of cerebral ischemia-reperfusion injury on glucuronidation metabolism of IMM-H004 in the brain
摘要
Abstract
OBJECTIVE To investigate the impact of cerebral ischemia-reperfusion(CIR)injury on glucuronidation of IMM-H004 in the brain.METHODS IMM-H004,a neuroprotective agent,underwent glucuronidation primarily mediated by uridine diphosphate glucuronosyltransferases(UGT)to form IMM-H004G,which was subsequently hydrolyzed back to IMM-H004 by β-glucuronidase.① Cellular experiments:human glial cells(HEB)and human neuroblastoma cells(SH-SY5Y)cell lines were assigned to two groups:a normal control group and an oxygen-glucose deprivation/reoxygenation(OGD/R)model group.An OGD/R model was established by subjecting the cells to one-hour oxygen-glucose deprivation,followed by reoxygenation.Cell viability was assessed using the methylthiazolyldi-phenyl-tetrazolium bromide(MTT)assay.The mRNA levels of UGT and its regulatory factor,nuclear factor erythroid 2 related factor 2(Nrf2),were measured by real-time fluorescence quantitative PCR(RT-qPCR).The content of IMM-H004G glucuronidated from IMM-H004,and the content of IMM-H004 hydrolyzed from IMM-H004G were determined using liquid chromatography-tandem mass spectrometry(LC-MS/MS).② Animal experiments:Male SD rats were randomly assigned to three groups:a normal control group,a CIR model group,and a sham operation group.Rats in the normal control group received no surgical interventions while those in the CIR model group underwent four-vessel occlusion surgery to induce acute CIR injury.Rats in the sham operation group was treated the same way as the CIR model group except for the four-vessel occlusion.The activities of UGT and β-glucuronidase in brain tissues were determined by LC-MS/MS.IMM-H004 was administered via intracerebroventricular injec-tion,and the concentrations of IMM-H004 and IMM-H004G in different regions of the brain were deter-mined using LC-MS/MS to investigate the impact of CIR injury on IMM-H004 metabolism.RESULTS① Cellular experiments:Compared with the control group,OGD/R injury reduced the viability of HEB and SH-SY5Y cells to 72.30%and 53.56%,respectively.In HEB cells,OGD/R injury significantly down-regulated the mRNA expressions of UGT1A1,UGT1A7 and UGT1A8,resulting in a reduction of IMM-H004G production to 50.05%-68.95%of the normal level,while hydrolytic metabolism remained unaf-fected.No significant changes were observed in SH-SY5Y cells.②Animal experiments:CIR injury had no impact on the activity of UGT or β-glucuronidase in rat brain tissues.In addition,the distribution of IMM-H004 and IMM-H004G across different brain regions remained unchanged.CONCLUSION These findings show that OGD/R injury reduces UGT-mediated glucuronidation of IMM-H004,whereas CIR injury does not significantly affect its metabolism in the brain,suggesting the presence of compen-satory mechanisms in brain tissues that help maintain drug homeostasis.关键词
脑缺血再灌注/尿苷二磷酸葡萄糖醛酸转移酶/β葡萄糖醛酸苷酶/神经胶质细胞/IMM-H004/IMM-H004GKey words
cerebral ischemia-reperfusion/uridine diphosphate glucuronosyltransferases/β-gluc-uronidase/neuroglial cell/IMM-H004/IMM-H004G分类
医药卫生引用本文复制引用
张伟琳,张梓倩,孙韬,李燕,盛莉..脑缺血再灌注损伤对脑内IMM-H004葡萄糖醛酸化代谢的影响[J].中国药理学与毒理学杂志,2025,39(7):489-499,11.基金项目
中国医学科学院医学与健康科技创新工程(2021-I2M-1-028) Chinese Academy of Medical Sciences Medical and Health Science and Technology Innovation Project(2021-I2M-1-028) （2021-I2M-1-028）
