| In recent years,the fluorescence imaging technology based on small molecular fluorescent probe has been developed rapidly,and it has been widely used in chemistry,biology,medicine and other fields.A large number of fluorescent probes have been developed for the detection of biological-related molecules,ions and proteinases,which are powerful tool for the study of cell physiology or pathological process.Among them,hypochlorous acid in lysosome has attracted extensive attention and research interest because of its role in physiological and pathological process.Hypochlorous acid is one of reactive oxygen species in organism,which is produced by MPO catalysis of H2O2and Cl-reaction in phagocytes.Lysosome is an important organelle in mammalian cells,which is responsible for the degradation and recovery of macromolecules,which plays a key role in maintaining the normal operation of cells.Lysosome is one of the main producing sites of hypochlorous acid.Abnormal level of HClO in lysosomes can cause cell damage,and lead to many diseases,such as Parkinson disease,cardiovascular disease,arthritis and cancer.Therefore,it is of great significance to develop a high sensitivity and high selectivity fluorescent probe for the monitoring of the concentration and distribution of HClO in lysosomes,and also to the research of physiological and pathological processes related to HClO in lysosomes,and thus provide theoretical basis for the prevention and early diagnosis of diseases.Based on the above reasons,two lysosome-targetable fluorescent probes for hypochlorous acid were designed and synthesized,and the specific contents are as follows:(1)In Chapter 2,a lysosome-targetable fluorescent probe(Lyso-R-HClO)for hypochlorous acid was designed and synthesized based on Rhodamine B dye.Lyso-R-HClO showed high sensitivity and selectivity to hypochlorous acid.The linear response range of Lyso-R-HClO to hypochlorous acid was 5.0×10-8-6.0×10-6M,and the detection limit was 2.0×10-8M.In addition,Lyso-R-HClO has excellent lysosome-targetable ability and has been successfully applied to imaging exogenous and endogenous HClO in lysosome.Finally,we successfully used Lyso-R-HClO to monitor hypochlorous acid in macrophage inflammation.The results show that Lyso-R-HClO is a powerful tool to study the biological and pathological functions of HClO at the level of organelle.(2)In Chapter 3,a lysosome-targetable two-photon excited near-infrared fluorescent probe(Lyso-TP-HClO)for hypochlorous acid was designed and synthesized.Lyso-TP-HClO has high selectivity and sensitivity to hypochlorous acid,and its linear response range is 10.0×10-8-5.0-10-6M,and the detection limit is 3.0×10-8M.At the same time,Lyso-TP-HClO has good imaging performance,such as small autofluorescence,excellent photostability and large imaging depth.In addition,we successfully used Lyso-TP-HClO to detect the exogenous and endogenous HClO in lysosome.Finally,Lyso-TP-HClO was further applied to the visualization of HClO in macrophages infected by bacteria and in arthritis,and satisfactory results were obtained.All the results show that Lyso-TP-HClO can be used in the imaging of lysosome HClO and the diagnosis of early arthritis,and it is a powerful tool for the detection of HClO. |
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