首都医科大学学报 ›› 2025, Vol. 46 ›› Issue (1): 76-82.doi: 10.3969/j.issn.1006-7795.2025.01.013

• 医学图像分析在心脑疾病诊断及预测中的应用 • 上一篇    下一篇

利用BIBP-H荧光探针进行脑缺血再灌注损伤检测

崔  玮1#,  庞淇丹2,3#,  相韩悦4,  肖  猱4,  姜德春2,3,  李  深1,  沈光莉1*   

  1. 1.首都医科大学附属北京世纪坛医院神经与精神科,北京 100038;2.首都医科大学药学院临床药学系,北京 100069;3.首都医科大学附属北京世纪坛医院药学部,北京 100038;4.首都医科大学药学院化学生物学系,北京 100069
  • 收稿日期:2024-10-24 出版日期:2025-02-21 发布日期:2025-02-25
  • 通讯作者: 沈光莉 E-mail:shen_guangli@sina.com
  • 基金资助:
    国家自然科学基金项目(82371298, 82311530048),首都卫生发展科研专项(2024-2-2086),首都医科大学附属北京世纪坛医院领军人才培养项目(2024LJRCLS)。

Detection of cerebral ischemia-reperfusion injury using BIBP-H fluorescent probe

Cui Wei1#, Pang Qidan2,3#, Xiang Hanyue4, Xiao Nao4, Jiang Dechun2,3, Li Shen1, Shen Guangli1*   

  1. 1.Department of Neurology and Psychiatry, Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, China;2.Department of Clinical Pharmacy, School of Pharmaceutical Sciences, Capital Medical University, Beijing 100069, China;3.Department of Pharmacy, Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, China;4.Department of Chemical Biology, School of Pharmaceutical Sciences, Capital Medical University, Beijing 100069, China
  • Received:2024-10-24 Online:2025-02-21 Published:2025-02-25
  • Supported by:
    This study was supported by the National Natural Science Foundation of China (82371298, 82311530048), Capital's Funds for Health Improvement and Research (2024-2-2086), Leader Talent Training Program of Beijing Shijitan Hospital, Capital Medical University (2024LJRCLS).

摘要: 目的  明确BIBP-H荧光探针是否可以用于脑缺血再灌注后氧化应激水平的检测。方法  首先,在体外利用BIBP-H探针荧光成像检测谷氨酸诱导的大鼠神经母细胞瘤细胞(B104)氧化应激反应,并观察抗氧化剂依达拉奉右莰醇或还原型谷胱甘肽预处理对B104细胞氧化应激后荧光强度的影响。随后,将28只健康雄性C57BL/6小鼠采用抽签法随机分为短暂大脑中动脉栓塞(transient middle cerebral artery occlusion, tMCAO)、依达拉奉右莰醇+tMCAO、还原型谷胱甘肽+tMCAO和假手术4组(每组7只),缺血1.5 h,再灌注12 h后,经尾静脉注射BIBP-H探针,利用活体成像及后续离体和组织荧光成像进行BIBP-H探针的体内缺血再灌注损伤检测。结果  ①仅给予BIBP-H探针的B104细胞未见自发性红色荧光;BIBP-H探针可在谷氨酸刺激的B104细胞中显示红色荧光,依达拉奉右莰醇或还原型谷胱甘肽预处理后则荧光强度显著下降;②静脉注射BIBP-H探针的假手术小鼠和未注射探针的tMCAO小鼠均呈活体荧光成像阴性;③静脉注射BIBP-H探针的tMCAO小鼠病灶区呈活体荧光成像阳性,且再灌注同时给予依达拉奉右莰醇和还原型谷胱甘肽静脉输注后病灶区的荧光强度下降;④BIBP-H探针显示的红色荧光范围与脑梗死范围一致。结论  BIBP-H探针能有效监测体内外氧化应激反应,在脑缺血再灌注损伤检测中具有应用前景。

关键词: 荧光探针, 脑缺血再灌注损伤, 氧化应激, 生物成像

Abstract: Objective  To evaluate the potential of the BIBP-H fluorescent probe in the detection of  the oxidative stress levels after cerebral ischemia-reperfusion (CIRI). Methods  In vitro, the potential of  BIBP-H probe was in detection of oxidative stress was first assessed with fluorescence imaging in rat neuroblastoma (B104) cells after L-glutamic acid stimulation. And then, the effects of edaravone and dexborneol (EDA) and glutathione (GSH) pretreatment on the fluorescence intensity were evaluated. Later, a totally of 28 male C57BL/ 6 mice were randomly assigned into four groups: transient middle cerebral artery occlusion (tMCAO) group, EDA+tMCAO group, GSH+tMCAO group, and sham group. After 1.5 h ischemia and 12 h  reperfusion, the mice were treated with BIBP-H via tail vein injection. In vivo, ex vivo, and tissue fluorescence imaging were utilized to evaluate the probe's cerebral ischemia - reperfusion injury (CIRI).Results  ① BIBP-H probe did not exhibit fluorescence signals in cultured B104 cells, but showed red fluorescence in B104 cells treated with L-glutamic acid. The signals significantly decreased when pretreated with EDA or GSH. ② Both the sham-operated mice intravenously injected with the BIBP-H probe and the tMCAO mice without injection of the probe showed negative results in in vivo fluorescence imaging.  ③ tMCAO mice treated with BIBP-H exhibited red fluorescence signals in the ischemic hemisphere in vivo, with significantly reduced fluorescence intensity after EDA or GSH infusion during reperfusion  ④ The fluorescence area examined with BIBP-H was consistent the cerebral infarction area detected with triphenyltertrazolium. Conclusion  The BIBP-H probe effectively monitored oxidative stress levels both in vivo and in vitro, demonstrating its potential in CIRI detection.

Key words: fluorescent probe, cerebral ischemia/reperfusion, oxidative stress, biological imaging

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