Design, Preparation And Application Of Peroxynitrosyl Anion And Nitrosothiol In Situ Fluorescence Imaging Probe

Intracellular reactive nitrogen?RNS?refers to NO and molecules derived from NO.The oxidation state of nitrogen element is+1 to+4,such as peroxynitrosyl anion?ONOO^-?,nitrogen dioxide?NO₂?.S-nitrososterol?RSNO?and nitrosyl hydrogen?HNO?.RNS is capable of reacting with a variety of biomolecules including proteins,nucleic acids and lipids.Among them,ONOO^- is produced by the reaction of nitric oxide?NO?and superoxide anion radical?·O₂^-?,and its abnormal fluctuation may cause oxidative stress and nitrative stress,which in turn is associated with various diseases.RSNO is an intermediate product in the process of nitroso metabolism in vivo.The trans-nitroso process between it and nitrosoprotein has an important effect on the functional expression of proteins and enzymes,and has important physiological effects in organisms.At present,the most interesting aspect of RSNO is its role of NO pool reservior and NO transport.In view of the important physiological functions of ONOO^- and RSNO,it is particularly important to construct an analytical method capable of measuring ONOO^- and RSNO in situ.Based on this,two fluorescent probes of TPE-COU and NIR-GSNO were designed and synthesized in this paper which were used for fluorescence detection of ONOO^- and RSNO,respectively.The specific work is as follows:?1?Based on the mechanism of aggregation induced emission?AIE?,a probe of TPE-COU was constructed for ONOO^-.TPE-COU was fabricated by linking a bromo-coumarin derivative to a TPE-based motif through a?,?-unsaturated ketene,which was served as a recognition site for ONOO^-.After?,?-unsaturated ketene is decomposed by ONOO^-,AIE is triggered by forming the free tetrastyrene moiety in the aqueous solution.Combined with a H₂S specific probe of azidocoumarin,TPE-COU was applied to the detection of ONOO^-in cells during oxyglucose deprivation reperfusion?OGD/R?.It was proved that estrogen protects cardiomyocytes from oxidative stress during OGD/R by upregulating endogenous H₂S and the level of ONOO^- was decreased.The results showed that ONOO^- can be used as an indicator in the process of myocardial injury,and TPE-COU has a good application potential in the related physiological and pathological research.?2?Based on the mechanism of intramolecular charge transfer?ICT?,a near-infrared probe NIR-GSNO was designed to study the changes of S-nitrosothiol content during NO photoreleasing.The important role of biothiol in the storage and transportation of NO is illustrated.The results provide useful analytical methods and related data for an in-depth understanding of the physiological and pathological functions of NO and nitrosothiol.