PDF(Pubmed)
Abstract：
Two NAD(P)H-biosensing probes consisting of 1,3,3-trimethyl-3H-indolium and 3-quinolinium acceptors, linked by thiophene, A, and 3,4-ethylenedioxythiophene, B, bridges are detailed. We synthesized probes C and D, replacing the thiophene connection in probe A with phenyl and 2,1,3-benzothiadiazole units, respectively. Probe E was prepared by substituting probe A\'s 3-quinolinium unit with a 1-methylquinoxalin-1-ium unit. Solutions are non-fluorescent but in the presence of NADH, exhibit near-infrared fluorescence at 742.1 nm and 727.2 nm for probes A and B, respectively, and generate absorbance signals at 690.6 nm and 685.9 nm. In contrast, probes C and D displayed pronounced interference from NADH fluorescence at 450 nm, whereas probe E exhibited minimal fluorescence alterations in response to NAD(P)H. Pre-treatment of A549 cells with glucose in the presence of probe A led to a significant increase in fluorescence intensity. Additionally, subjecting probe A to lactate and pyruvate molecules resulted in opposite changes in NAD(P)H levels, with lactate causing a substantial increase in fluorescence intensity, conversely, pyruvate resulted in a sharp decrease. Treatment of A549 cells with varying concentrations of the drugs cisplatin, gemcitabine, and camptothecin (5, 10, and 20 μM) led to a concentration-dependent increase in intracellular fluorescence intensity, signifying a rise in NAD(P)H levels. Finally, fruit fly larvae were treated with different concentrations of NADH and cisplatin illustrating applicability to live organisms. The results demonstrated a direct correlation between fluorescence intensity and the concentration of NADH and cisplatin, respectively, further confirming the efficacy of probe A in sensing changes in NAD(P)H levels within a whole organism.
摘要：
两个由1,3,3-三甲基-3H-吲哚和3-喹啉受体组成的NAD（P）H-生物传感探针，通过噻吩连接，A,和3,4-亚乙基二氧噻吩，B,桥梁是详细的。我们合成了探针C和D，用苯基和2,1,3-苯并噻二唑单元代替探针A中的噻吩连接，分别。通过用1-甲基喹喔啉-1-鎓单元取代探针A的3-喹啉鎓单元来制备探针E。溶液是非荧光的，但在NADH的存在下，探针A和B在742.1nm和727.2nm处表现出近红外荧光，分别,并在690.6nm和685.9nm处产生吸光度信号。相比之下,探针C和D在450nm处显示出来自NADH荧光的明显干扰，而探针E对NAD(P)H的反应表现出最小的荧光变化。在探针A存在下用葡萄糖预处理A549细胞导致荧光强度显著增加。此外,将探针A与乳酸盐和丙酮酸盐分子接触会导致NAD（P）H水平发生相反的变化，乳酸引起荧光强度的大幅增加，相反,丙酮酸导致急剧下降。用不同浓度的药物顺铂治疗A549细胞，吉西他滨,和喜树碱（5，10和20μM）导致细胞内荧光强度的浓度依赖性增加，表示NAD(P)H水平上升。最后,用不同浓度的NADH和顺铂处理果蝇幼虫，说明对活生物体的适用性。结果表明，荧光强度与NADH和顺铂的浓度之间存在直接相关性，分别,进一步证实探针A在感测整个生物体内NAD(P)H水平变化中的功效。
