Apigenin is a Potential Drug for Treating Ischemic Stroke. Association between Plasma Protein Genes and Ischemic Stroke: A Proteome-wideMendelian Randomization Analysis and Experimental Study

Introduction: Earlier research observed the effects of plasma proteins on ischemic stroke (IS). This Mendelian randomization (MR), in vivo study additionally assesses the associations of localized populations of plasma proteins with IS to further confirm the causality and explore drug targets. Method: This investigation consists of a Mendelian randomization assessment, an in vivo experiment, and molecular docking and molecular dynamics simulations. We mined data from the protein quantitative trait loci (pQTL) data from the deCODE Genetics Consortium and the IS outcome data from the MiBioGen consortium to identify 233 proteins from 4907 circulating proteins. We then performed MR analysis using the 233 proteins for the causal associations with IS risk. We developed an MCAO model in SD rats. Transcriptomic sequencing was performed on differentially expressed genes associated with plasma proteins that have causal relationships with IS, and the results were validated by real-time quantitative PCR. Further, candidate therapeutic agents are investigated on the basis of target screening, further molecular docking, and molecular dynamics simulations. Result: We thoroughly evaluated the impact of 233 plasma proteins on the risk of IS, finding that 26 of these proteins showed a potential association with the risk of outcome (P < 0.05). These proteins include PPP1R14A, IGF1R, PLG, PCDH10, IL15RA, RAB26, KNG1, HTATIP2, POMC, LILRA5, LYVE1, WISP1, LMAN2, NRXN1, TMEM9, HMBS, IL5RA, UNC5B, PTH, PUF60, STOML2, TFRC, KLRG2, VBP1, and HTATIP2. Transcriptomics and Mendelian randomization have identified overlapping targets, including Ppp1r14a, Stoml2, Lyve1, WISP1, IL5RA, Tfrc, and Klrg2. This suggests that these targets may play a significant role in the prediction and identification of IS. Apigenin, Metformin, Vitamin K 3, and Estradiol were potential therapeutic drugs. Molecular docking and molecular dynamics results show that TFRC has a tight binding with these drugs. Discussion: This MR study demonstrated IS associations for 26 plasma proteins, including 7 which could serve as biomarkers of IS: Ppp1r14a, Stoml2, Lyve1, WISP1, IL5RA, Tfrc, and Klrg2. These key targets' functional roles in the pathogenesis of IS were further ascertained. Furthermore, molecular docking and molecular dynamics simulations have shown that TFRC binds strongly to apigenin, metformin, menadione, and estradiol, which may suggest a mechanism of action of apigenin target TFRC. Conclusion: The key genes that are responsible for the occurrence and development of ischemic stroke are Ppp1r14a, Stoml2, Lyve1, WISP1, IL5RA, Tfrc, and Klrg2. Apigenin may be a Tfrctargeting agent for ischemic stroke treatment. Nonetheless, more in vivo and in vitro investigations are necessary to verify the mechanisms and the beneficial effects of apigenin interfering with Tfrc for the treatment of ischemic stroke.