Chemiluminescence Small Animal Imaging Study Of Disease Markers

Chemiluminescence imaging has been recognized as one of the most sensitive non-invasive imaging technique due to the advantage of dark field measurement.It has great application potential in basical medicine,clinical research,and drug development.Now,chemiluminescence imaging of living animals focuses on the direct determination of the reactive oxygen species(ROS)or the macromolecules related to the regulation of reactive oxygen metabolism.However,there is no report on the direct detection of protein,nucleic acid and other macromolecules,especially the biomarkers of multiple serious diseases via chemiluminescence imaging.On the one hand,medical andpharmaceutical research needs highly sensitive methods;on the other hand,chemiluminescence imaging is impeded by the limited object.Therefore,it is significant to detect protein markers besides ROS in vivo by chemiluminescence imaging.A highly efficient chemiluminescence probe,5-diformylhydrazinefluorecein,was synthesised in our previous work.Small animal experiments show that the probe can locate and detect horseradish peroxidase(HRP)enriched region in vivo.Here,by taking advantage of target recognification of antibodies labeled with HRP and protein,the HRP would be detected by chemiluminescence imaging,which means the protein markers could be determined.We envisage that the proposed method will offer a new perspective and innovative technology platform for pathology,pharmacology,and disease diagnosis in vivo.This paper realizes the detection of protein markers,such as vascular endothelial growth factor(VEGF),alpha-fetoprotein(AFP),tumor necrosis factor-a(TNF-a)in vivo,and then detects the tumor markers vascular endothelial growth factor receptor 2(VEGFR2)in vivo.Using VEGFR2 as the target,the target imaging of transplanted tumor in mice was realized.To explore the potential of chemiluminescence imaging in pharmacology and pharmacodynamics,inflammation was used as a template model.The frequency of administration and the efficacy of dexamethasone were preliminarily characterized in vivo.The above results varify the originality of our ideas and expand the application and technical methods of chemiluminescence imaging.The thesis covers the following aspects:Chapter 1:Overview.In this chapter,the basic principles of bioluminescence imaging and chemiluminescence imaging,the types and applications of imaging probes,the application and research of bioluminescence imaging and chemiluminescence imaging in recent decades,and the characteristics of the two kinds of imaging have been summarized and evaluated.Chapter 2:Detecting the interesting protein by chemiluminescence imaging.Three different protein,including VEGF,AFP,and TNF-a were chosen as templet.These protein have been imaged in nude mice by using HRP labeled antibody and 5-diformylhydrazinefluorecein as identified probe and chemiluminescence imaging probe respectively.The targeted protein was injected subcutaneously in nude mice.Next,the target recognition would be completed by the interaction between the targeted protein and the antibody labeled with HPR,so the purpose of detecting the interesting protein in vivo could be achieved.Chapter 3:Tumor imaging based on tumor marker imaging.We are encouraged to focus on the detection of protein markers in specific disease models,owing to the successful example of sensitive detection of protein in nude mice by chemiluminescence imaging.VEGFR2,tumor cell membrane protein marker,was selected as the target.The anti-VEGFR2 labeled HRP was used as the identified and report probe,and has successfully targeted the tumor issue in nude mice which were subcutaneously transplanted with human colon cancer cell line HCT-116.Chapter 4:Evaulating the efficacy of anti-inflammatory drug dexamethasone.Inflammation is associated with various major diseases,such as cancer,cardiovascular disease,and Alzheimer,and poses a threat on people's life.In our previous work,we have achieved a highly sensitive detection of inflammation by chemiluminescence imaging which has been verified a powerful tool applied in characterization of inflammation.The acute arthritis in mice was used as disease model to assess the efficiency of dexamethasone for exploring the potential of chemiluminescence imaging in development of anti-inflammatory drugs,pharmacology,and new drug.The frequency and efficacy of dexamethasone were characterized in vivo,and the chemiluminescence signals before and after administration were compared.A series of results show that evaluation of the efficacy of anti-inflammatory drugs by chemiluminescence imaging in vivo could be achieved.Chapter 5:Summary and outlook.