Atherosclerosis (AS) is a progressive inflammatory illness with extremely advanced pathogenesis, and it underlies main cardiovascular and cerebrovascular occasions resembling myocardial infarction, ischemic cardiomyopathy, and stroke, making it a number one explanation for morbidity and mortality worldwide [1], [2], [3], [4]. Amongst varied threat components, epidemiological and mechanistic research have recognized homocysteine (Hcy) as an impartial and demanding contributor to AS [5], [6]. Elevated Hcy damages endothelial cells and participates in a number of levels of plaque initiation, development, and rupture [7], [8], [9]. In parallel, oxidative stress has additionally been acknowledged as a central driver of atherosclerotic lesions [10], [11]. Below oxidative stress, the intracellular microenvironment undergoes profound adjustments, together with irregular viscosity, which might alter metabolite diffusion, impair sign transduction, and exacerbate mobile dysfunction [12], [13]. Regardless of its potential significance, the position of viscosity dysregulation in AS stays poorly understood, and whether or not Hcy straight influences viscosity in atherosclerosis-related cells, resembling foam cells, is basically unexplored. Addressing this hole requires instruments that may concurrently monitor Hcy and viscosity dynamics in actual time.
Fluorescent probes have emerged as highly effective instruments for biomedical analysis owing to their excessive sensitivity, operational simplicity, and non-invasive nature [14], [15], [16], [17], [18], [19], [20], [21], [22], [23], [24]. A number of probes have been reported for AS-related research, primarily specializing in lipid droplets or reactive oxygen species (Desk S2) [25], [26], [27], [28], [29], [30], [31], [32], [33], [34], [35]. Nonetheless, just a few fluorescent probes have been tailor-made for selective detection of Hcy, attributable to its structural and chemical similarity to cysteine, and their utility in AS stays uncommon [36], [37], [38]. Moreover, probes concentrating on a single analyte typically fail to seize the multifactorial nature of AS. In distinction, dual-channel probes able to simultaneous and orthogonal detection of a number of analytes can ship extra complete mechanistic insights and improve diagnostic accuracy [39]. But many standard dual-channel techniques undergo from spectral overlap or sign interference, which compromises quantitative reliability in advanced organic environments. On this regard, spectrally orthogonal fluorescent probes provide distinct benefits: by using totally separated emission channels with out cross-talk, they permit exact, high-fidelity, and simultaneous monitoring of a number of targets in residing techniques, thereby overcoming a key limitation of conventional dual-channel probes.
Right here, we report the design and synthesis of a spectrally orthogonal fluorescent probe (HV) that allows real-time, dual-channel imaging of Hcy and viscosity in residing techniques. Probe HV was constructed by covalently integrating a selective Hcy-responsive module [40] with an environment-sensitive unit via a piperazine linker, attaining two totally separated emission channels with out spectral cross-talk. This design permits extremely selective, simultaneous monitoring of biochemical and biophysical adjustments in atherosclerosis-related cells. Utilizing HV, we uncovered a direct relationship between Hcy accumulation and viscosity dysregulation throughout foam cell formation induced by oxidized low-density lipoprotein (Ox-LDL). Furthermore, utility of HV in atherosclerotic mouse fashions revealed that decreasing Hcy ranges with folic acid and vitamin B12 supplementation alleviates viscosity abnormalities and attenuates illness development. These findings not solely set up a mechanistic hyperlink between Hcy, viscosity, and AS pathogenesis but additionally place HV as a strong platform for early analysis, mechanistic exploration, and therapeutic monitoring of atherosclerosis.
