Nobiletin protected against hypertrophic cardiomyopathy via targeting PPARα

Kewei Zhou^1,2Chang Chen^2,3Hexin Cai^2,3Zuqian Lian^2,3Luping Wang^2,3Qinghuo Li^1,2Xiaoqian Wu^2,4*Panxia Wang^2,4*

• ¹Department of Pharmacology, Guangzhou Medical University, Guangzhou, China
• ²Department of Pharmaceutical Science, School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou, China
• ³Depaertment of Pharmacology, Guangzhou Medical University, Guangzhou, China
• ⁴Department of Pharmaceutical Science, Guangzhou Medical University, Guangzhou, China

Hypertrophic cardiomyopathy is an independent risk factor for heart failure. Citrus reticulata (C. reticulata) is a traditional Chinese medicine with a variety of chemical components and pharmacological effects. The mechanisms of C. reticulata for treating hypertrophic cardiomyopathy is still unclear. In this study, we used network pharmacology techniques combined with bioinformatic analysis and identified the active ingredient in C. reticulata to protect against hypertrophic cardiomyopathy. Moreover, we further analyzed the GEO database from human heart tissue with hypertrophic cardiomyopathy to reveal the potential targets. Finally, molecular docking and in vitro validation were used to reveal the binding of the potential targets and the main active component of C. reticulata. Our results revealed that there are five main active ingredients of C. reticulata (Nobiletin, Naringenin, Sitosterol, 5,7-dihydroxy-2-(3-hydroxy-4-methoxyphenyl) chroman-4-one, and Citromitin). By analyzing the intersective genes between C. reticulata and hypertrophic cardiomyopathy, 40 hub genes were obtained. GO and KEGG analysis exhibited responses to oxidative stress and fatty acids were the main pathways for C. reticulata and hypertrophic cardiomyopathy. The protein-protein interaction analyzing results showed that the main active ingredients of C. reticulata were Nobiletin and Naringenin, while PPARα might be the potential targets of C. reticulata in treating hypertrophic cardiomyopathy. The molecular docking results showed that the main active ingredient Nobiletin could bind to PPARα with a strong hydrogen bonding interaction force. In vitro results validated that Nobiletin might directly bind to PPARα, thereby increasing the expression of lipid metabolism related genes and relieved hypertrophic responses of cardiomyocytes. In conclusion, the nuclear receptor PPARα might be the potential endogenous receptor of the active ingredients of C. reticulata.