Activation of the Nrf2 Antioxidant Pathway by Longjing Green Tea Polyphenols in Mice Livers

Previous studies have revealed that green tea polyphenol (GTP) could protect against liver injury due to oxidative stress. However, the mechanism underlying the bioactive actions of GTP in the liver has not been systematically evaluated. This study aimed to investigate the effect of GTP on the activation of the nuclear factor erythroid-2-related factor 2 (Nrf2)-Kelch-like ECH-associated protein 1 (keap1) pathway, using in silico and in vivo methods. Furthermore, the regulation of Nrf2 downstream target antioxidant response element (ARE) was also evaluated. The high-performance liquid chromatography analysis indicated that GTP includes 9 major compounds, and molecule docking analysis demonstrated that most of these polyphenols have a strong binding affinity with the keap1 Kelch domain, where keap1 binds to the Neh2 domain of Nrf2. Remarkably, the predominant compound of GTP, that is, epigallocatechin gallate, displayed the best binding affinity score, which can fully occupy all 3 polar subpockets of the keap1 Kelch domain. The Nrf2, keap1, and Nrf2 downstream target gene expression levels were changed in the livers compared to the control group. It showed that the Nrf2 expression level was significantly upregulated in GTP-induced mice liver across most treatments, while the keap1 expression level remained unchanged. Subsequently, we observed a significant increasing trend in the expression of the downstream ARE, including antioxidative enzymes, liver phase II enzymes, and liver efflux transporters in mice livers. The present study demonstrated that GTP could activate the Nrf2 signaling pathway by interrupting the Nrf2-keap1 protein–protein interaction