Liensinine Ameliorating Oxidative Damage to Vascular Endothelium In Vitro via Upregulating eNOS and Nrf2/HO-1...

Liensinine, as a main alkaloid from Plumula nelumbinis, shows a significant effect against vascular-related diseases, especially oxidative damage to vascular endothelium (ODVE). However, the underlying mechanism is unknown. The targets of liensinine against ODVE, the binding energies and molecular dynamics simulation between the proteins and liensinine were researched in the present study. The result was validated by protecting H₂O₂-injured HUVECs via measuring cell viability, migration, apoptosis, contents of ROS and NO, expressions of Nrf2, HO-1, SOD2, eNOS and p-eNOS, and Nrf2 localization. SNP, L-NAME, and ML385 were used as NO donor, eNOS inhibitor and Nrf2 inhibitor to validate the effect of liensinine in regulating Nrf2/HO-1 pathway and eNOS activity. Venn analysis identified 77 common targets between liensinine and ODVE. According to the results of PPI analysis and molecular docking, liensinine possessed strong affinities to NFKB1, NFE2LE, and ESR1. Molecular dynamics simulation result indicated that NFE2LE-liensinine was the most stable complex. GO and KEGG enrichment results indicated that NO generation and oxidative stress were the main mechanisms involved in the protection. In the in vitro validation experiment, liensinine exhibited a strong protective effect against ODVE, including enhancing cellular activities, migration and NO generation, reducing ROS production and cellular apoptosis. The underlying mechanism might be associated with the upregulation of eNOS and Nrf2/HO-1 pathway, which may even exhibit cross-talk modulation. These findings reveal that liensinine has a strong potential to be used as a dietary supplement for patients with injured vascular endothelium due to oxidative stress.