Study on rhynchophylline reducing vascular endothelial cells prethrombotic state induced by TNF-α through enhancing autophagy / 中草药

ABSTRACT

Objective: To investigate the effect of rhynchophylline on HUVECs prethrombotic state induced by TNF-α through regulating autophagy. Methods: TNF-α was used to establish the HUVECs damage model. The change of autophagy was detected by transmission electron microscope and immunofluorescence technique. Western blotting was used to detect the LC3II/LC3I ratio and the expression of Beclin-1, p62, NF-κB, vWF, and PAI-1. The content of ET-1 and PGI2 in cultural supernatant was detected by ELISA. Results: TNF-α up-regulated the expression of NF-κB, vWF, and PAI-1in HUVECs, and decreased the content of ET-1 and PGI2 in cell culture supernatant (P < 0.05). In addition, more autophagosomes were observed in HUVECs, and the LC3II/LC3I ratio and beclin-1 expression were increased, as well as the expression of p62 was decreased (P < 0.05). Rhynchophylline promoted the production of autophagosome, increased the LC3II/LC3I ratio and Beclin-1 expression, and reduced the expression of p62 (P < 0.05). Moreover, the results showed that rhynchophylline down-regulated the expression of NF-κB, vWF, and PAI-1, as well as reduced the content of ET-1 and PGI2 in cultural supernatant (P < 0.05). 3-MA, an inhibitor of autophagy, abolished the effect of rhynchophylline to a certain extent (P < 0.05). Conclusion: Rhynchophylline reduces the expression of coagulative factors induced by TNF-α through enhancing autophagy, and inhibits the prethrombotic state when vascular endothelial cells have inflammatory damage.