器官移植2026，Vol.17Issue(5)：783-794,12.DOI:10.12464/j.issn.1674-7445.2026149
丹参注射液通过Nrf2通路减轻人脑类器官氧化损伤的机制研究
Research on the mechanism of Danshen injection in reducing oxidative damage in human brain organoid via the Nrf2 pathway
摘要
Abstract
Objective To investigate whether Danshen injection(DSI)can protect human brain organoids from oxidative stress damage by regulating the Nrf2 signaling pathway and its mechanism.Methods A Nrf2 plasmid was constructed,lentivirus was packaged and transfected into human induced pluripotent stem cell(hiPSC)to form a stable cell line.Brain organoids with Nrf2 overexpression(LV-Nrf2 group)and knockdown(sh-Nrf2 group)were constructed using kit.The transfection efficiency was verified by real-time fluorescence quantitative polymerase chain reaction.An oxidative stress injury model was established by inducing hydrogen peroxide(H2O2).Intervention was performed either in combination or without DSI pre-treatment.Cell viability was detected by Cell Counting Kit-8(CCK-8),and the expression levels of Nrf2,malondialdehyde(MDA),interleukin(IL)-1β,sex-determining region Y-box protein 2(SOX2)and neuronal-specific nuclear protein(NeuN)were detected by Western blot and immunofluorescence staining.Total RNA from human brain organoids was extracted for transcriptome sequencing and analysis.Results LV-Nrf2 group and sh-Nrf2 group brain organoid models were successfully constructed.The CCK-8 results determined that the oxidative stress model was established with 32 mmol/L H2O2 intervention conditions.The fluorescence expression of Nrf2 in LV-Nrf2 group was higher than that in control group,and the fluorescence expressions of MDA and IL-1β were lower than those in control group;sh-Nrf2 group showed a damage phenotype of low Nrf2 expression,high MDA and IL-1β expression(all P＜0.05).Compared with H2O2 group,the Nrf2 signal in DSI+H2O2 group was stronger,and the levels of MDA and IL-1β were lower.Among them,the protective effect of LV-Nrf2 was the most significant.H2O2 treatment could down-regulate the protein levels of Nrf2,SOX2 and NeuN,and DSI intervention could effectively reverse the downward trend of these indicators.Sh-Nrf2 could weaken the recovery effect of DSI on SOX2 and NeuN,while LV-Nrf2 could synergistically enhance the regulatory effect of DSI.Conclusions DSI may antagonize oxidative stress damage in human brain organoids through the Nrf2 signaling pathway,and maintaining the stability of the neural cell lineage is its core therapeutic characteristic.关键词
丹参注射液/人诱导多能干细胞/脑类器官/核因子E2相关因子2/氧化应激损伤/缺血-再灌注损伤/缺血性卒中/基因修饰Key words
Danshen injection/Human induced pluripotent stem cell/Brain organoid/Nuclear factor E2-related factor 2/Oxidative stress injury/Ischemia-reperfusion injury/Ischemic stroke/Gene modification分类
医药卫生引用本文复制引用
严紫娟,李冠东,赵子祯,王雪焱,陈雪瑜,吴润灵,穆淑花,王家传..丹参注射液通过Nrf2通路减轻人脑类器官氧化损伤的机制研究[J].器官移植,2026,17(5):783-794,12.基金项目
国家自然科学基金面上项目(82274248、81774053) （82274248、81774053）
深圳市"医疗卫生三名工程"项目(SZZYSM202411009) （SZZYSM202411009）
深圳市自然科学基金面上项目(JCYJ20220531092403007) （JCYJ20220531092403007）
深圳市医学研究基金(C2301009) （C2301009）
