Fluorescence probe for detecting intracellular hydrogen sulfide and preparation method and application of fluorescence probe

Abstract

The invention relates to a fluorescence probe for detecting intracellular hydrogen sulfide and a preparation method and application of the fluorescence probe. Cy7 fluorochrome and sodium acetate are dissolved in anhydrous dimethylformamide (DMF), heating reflux is carried out for 2-4 hours under protection of Ar gas, cooling and suction filtration are carried out on an obtained mixture, mother liquor is crystallized under the low temperature of minus 4-20 DEG C, and keto form cyanine reddish brown crystals are obtained after the suction filtration; toluoyl benzoic acid and oxalyl chloride are dissolved in anhydrous CH2Cl2, then a bit of DMF is added, mixing is carried out for 2-6 hours under the temperature of 0 DEG C, and toluoyl benzoyl chloride is obtained; and keto form cyanine and triethylamine are dissolved in the anhydrous CH2Cl2 and are cooled to be 0 DEG C, the toluoyl benzoyl chloride is added dropwise, mixing is carried out for 10-30 minutes under the temperature of 0 DEG C, the mixture is heated to be in the room temperature, mixing is carried out overnight, suction filtration and spinning drying are carried out, separation is carried out through a chromatographic column, green solids are obtained after the spinning drying, and the structural formula is as follows. The fluorescence probe is simple in composition, high in detection speed, free of long-term incubation in a detection process, high in sensitivity, and capable of detecting intracellular endogenic hydrogen sulfide.

Description

Technical field: The invention relates to a fluorescent probe, in particular to a cyanine dye fluorescent probe, its synthesis method and its application in detecting intracellular hydrogen sulfide, belonging to the technical field of detection. Background technique: For a long time, hydrogen sulfide (H 2 S) This gas has the smell of rotten eggs. For many years, people have thought that H 2 S is only a poisonous gas, and people's research on it is limited to its toxic effect. In the mid-1990s, it was discovered that the body can endogenously produce H 2 S gas, which is mainly catalyzed by cystathionine-β-synthase (CBS) and cystathionine-γ-lyase (CSE) in the body to catalyze L-cysteine Acid produces H 2 S. CSE genes are expressed in the aorta, pulmonary artery, mesenteric artery, tail artery and portal vein, while CBS genes are mainly expressed in the central nervous system. h 2 S has the function of regulating nerve, cardiovascular and digestive system functions, and ...

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Problems solved by technology

Such as fluorescein as the parent (Hanjing Peng, YunfengCheng, Chaofeng Dai, Adrienne L. King, Benjamin L. Predmore, David J. Lefer, and Binghe Wang, Angew. Chem. Int. Ed. 2011, 50, 9672-9675) The probe has good selectivity, and the fluorescence is enhanced to 55-70 times before and after detection, but the reaction time of this probe is long (1h), and the excitation wavelength is in the visible light region (488nm); BODOPY is used as the matrix and (Yong Qian, Jason Karpus , Omer Kabil, Shu-Yu Zhang, Hai-Liang Zhu, Ruma Banerjee, JingZhao, Chuan He, Nature Communications.2011, 2, 495) with rhodamine as the parent (AlexanderR.Lippert, Elizabeth J.New, and Christopher J.Chang , J.Am.Chem.Soc., 2011,133:10078-10080) fluorescent probes, although the selectivity is good, but the reaction time is long (20min or 1h); some fluorescent probes for detecting hydrogen sulfide are dissolved due to their short wavelength Factors such as poor sex cannot be used in vivo, such as (Hanjing Peng, Yunfeng Cheng, Chaofeng Dai, Adrienne L. King, Benjamin L. Predmore, David J. Lefer, and Binghe Wang, Angew. Chem. Int. Ed. 2011, 50 , 9672-9675), although this probe is a fast response, but the selected dansyl is the parent, the excitation is 340nm, and the wavelength is short, if it is used in a living body, it will cause damage

In general, the above fluorescent probes are generally not water-soluble, and the excitation and emission wavelengths are mostly in the ultraviolet-visible region, which limits their application in vivo

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