靶向炎性脑水肿的诊疗一体化近红外荧光探针的构建与评价  

Construction and Evaluation of Theranostic Near-infrared Fluorescent Probe for Targeting Inflammatory Brain Edema

作　　者：秦靖 赵勇 张彩勤 白冰 师长宏 QIN Jing;ZHAO Yong;ZHANG Caiqin;BAI Bing;SHI Changhong(Laboratory Animal Center,Air Force Medical University,Xi'an 710032,China)

机构地区：[1]空军军医大学实验动物中心,西安710032

出　　处：《实验动物与比较医学》2024年第3期243-250,共8页Laboratory Animal and Comparative Medicine

基　　金：军队实验动物专项科研课题“靶向性近红外荧光探针在损伤性脑水肿模型评估与治疗中的应用”(SYDW〔2020〕21号);陕西省创新能力支撑计划“创伤性脑水肿模型的制备及影像学实时监测评估”(2022PT-45)。

摘　　要：目的制备基于近红外荧光(near-infrared fluorescence,NIRF)探针的新型化合物,通过活体成像实现对炎性脑水肿小鼠模型的动态监测,以及对治疗效果的实时评估。方法选择NIRF探针IR-783与临床脑水肿治疗药物呋塞米(furosemide,FSM)进行化学连接,获得新的化合物IR-783-FSM。通过紫外分光光度计评价该化合物的紫外荧光特性;体外细胞学实验检测小鼠巨噬细胞RAW 264.7对该化合物的摄取情况;CCK8实验评价该化合物的细胞毒性。BALB/c小鼠腹腔注射脂多糖构建炎性脑水肿模型,通过HE染色和测量脑组织干湿重法确认建模成功;将脑水肿模型小鼠分为对照组、IR-783和IR-783-FSM治疗组,分别给予PBS、IR-783和IR-783-FSM腹腔注射后进行实时活体荧光成像,并在10 h后处死各组小鼠,进行脑部离体成像和干湿重测量,观察IR-783-FSM对炎性脑水肿模型的NIRF成像特性和治疗效果。结果新合成的化合物IR-783-FSM保留了IR-783优良的近红外荧光特征,可以靶向小鼠巨噬细胞,IC50为48.82μmol/L。腹腔注射脂多糖可以成功构建炎性脑水肿模型,且其脑组织含水量较空白对照组小鼠明显升高(P<0.01);小鼠活体成像显示,与IR-783相比,IR-783-FSM在脑水肿模型中具有明显较强的荧光信号;与对照组相比,2、5和8 mmol/L IR-783-FSM治疗组小鼠的脑含水量均明显减少(P<0.01)。结论合成的新型NIRF探针IR-783-FSM可以实现对脑水肿的动态监测和对治疗效果的实时评估。Objective A novel compound based on near-infrared fluorescence(NIRF)probe was prepared to achieve dynamic monitoring of an inflammatory brain edema model in mice and real-time evaluation of therapeutic effects through invivoimaging.Methods The NIRF probe IR-783 was chemically linkedwith clinicalbrain edematherapeuticdrugfurosemide(FSM)toobtain the new compound,IR-783-FSM.The ultraviolet fluorescence properties of the compound were evaluated using an ultraviolet spectrophotometer.The uptake of the compound by mouse macrophage cells RAW 264.7 was detected with invitro cellular experiments.Its cytotoxicity was evaluated through CCK8 assays.A brain edema model was established in BALB/c mice via intraperitoneal injection of lipopolysaccharide(LPS),confirmed by HE staining and dry-wet weight methods for brain tissues.The mice in the brain edema model were divided into control group,IR-783,and IR-783-FSM treatment groups,receiving intraperitoneal injections of PBS,IR-783,and IR-783-FSM,respectively.Real-time invivofluorescence imaging was then performed.The mice in each group were euthanized after 10 hours.Ex vivo brain imaging and dry-wet weight measurements were performed to observe the NIRF imaging characteristics and therapeutic effects of IR-783-FSM on brain edema model.Results The newly synthesized compound,IR-783-FSM,retained the excellent near-infrared fluorescence characteristics of IR-783.It could target mouse macrophages with an IC50 of 48.82µmol/L.A brain edema model could be successfully constructed with intraperitoneal injection of LPS,with significantly higher brain tissue water content compared to the control group(P<0.01).Invivo imaging showed that IR-783-FSM had a significantly stronger fluorescence signal in the brain edema model than IR-783.Compared to the control group,the brain water content was significantly reduced in the 2,5,and 8 mmol/L IR-783-FSM treatment groups(P<0.01).Conclusion The newly synthesized NIRF probe IR-783-FSM facilitates dynamic monitoring of brain edema and real-time evaluati

关 键 词：脑水肿 脂多糖 近红外荧光 呋塞米 诊疗一体化 小鼠 

分 类 号：R-332[医药卫生] Q95-33[生物学—动物学]