Synthesis And Biological Application Of Fluorescent Probes For Intracellular HClO/NO Detection

Intracellular signal transduction and intercellular chemical communication depend largely on the control mechanisms associated with active species.Acitive species are generally considered to be indispensable for normal physiological functions,and the excess level caused by the increased production of active species or weakened antioxidant defense is related to inflammation,chronic cardiovascular disease,nervous system disease and other diseases.Therefore,it is important to design probes that can be used to detect Bio-Active Species.(1)Fluorescent dyes Rhodol-F and Rhodol isomer-F were designed synthesized by introducing strong electron-withdrawing halogen element F atom on the bottom ring with Rhodol as the parent structure.The introduction of four F atoms not only increases the transfer of intramolecular electrons,but also effectively redshifts the absorption wavelength and emission wavelength,and the F at the fourth position of the bottom ring has great reactivity.We constructed three fluorescent probes Rh-Se,Ri-Se1 and Ri-Se2 with reversible dynamic monitoring of redox by introducing phenyl Se groups.Through UV absorption spectroscopy and fluorescence emission on tests,we found that the probe Rh-Se exhibited good reversibility for Cl O~-/GSH detection.The detection limit of Cl O~-was 63.3 n M and the the detection limit of GSH was 71 n M.The probe Ri-Se1 can monitor the Cl O~-/Hcy redox process,and its deteciton limit for Cl O~-is 71 n M,and the detection limit for Hcy is 60 n M.The detection limits of Ri-Se2 for Cl O~-/Hcy were 0.13μM and 0.11μM respectively.(2)Based on the previous work,we constructed a fluorescent probe Rh-N with rhodamine as the parent structure by using ophenylenediamine to have a good recogniton mechanism for NO.The probe showed high selectivity and high sensitivity to NO in the water/acetonitrile 9:1 system.The maximum UV absorption wavelength redshifted from 470 nm to 510 nm after interaction with NO,which was mainly due to the fact that o-phenylenediamine acted with NO as the recognition site to form imidazole structure.The fluorescence emission wavelength redshifted from 594 nm to603 nm,and the fluorescence signal was turned on.The response time of probe Rh-N to NO was 6 hours,and the detection limit was 5.1 n M.In addtion,it is easy to react with o-phenylenediamine include ascorbic acid(AA),docosahexaenoic acid(DHA),carbonyl aldehyde(formaldehyde,acetone aldehyde)etc.Selective experiments found that the probe Rh-N had little effect on the above substances,and cell imaging experiments showed that the probe could achieve fluorescence imaging detection of exogenous and endogenous NO.