OS 32-07 LEVOROTATORY-AMLODIPINE PLUS FLUVASTATIN CONFER ADDITIVE EFFECTS ON RETARDING ENDOTHELIUM-MESENCHYMAL TRANSITION THROUGH SUPPRESSING ROCK1 EXPRESSION

To investigate the effects and mechanisms of levorotatory (L)-amlodipine combined fluvastatin treatment on endothelium-mesenchymal transition (EndMT), a process related to hypertension development. Human umbilical vein endothelial cells (HUVECs) were used and divided to six groups based on different therapeutic strategies as follows: Blank Control, angiotensin-II (Ang-II), Ang-II+ rho-associated kinase (ROCK) inhibitor, Ang-II+ L-amlodipine, Ang-II+ fluvastatin, and Ang-II +L-amlodipine + fluvastatin groups. Twenty-four hours later, HUVECs were collected for immunofluorescence staining and western blot analyses to evaluate EndMT, and for flow cytometry to analyze cells apoptosis. Compared to Blank Control group, ROCK1 expression was significantly increased and phosphorylated endothelial nitric oxide synthase (p-eNOS) and eNOS expressions were profoundly reduced in Ang-II group. Immunofluorescence staining revealed that CD31 expression was down-regulated and α-SMA expression was up-regulated in HUVECs in Ang-II group. Western blot showed that endothelium-specific markers (CD31 and VE-cadherin) were significantly diminished and fibroblasts-specific markers (α-SMA and FSP-1) were significantly increased by Ang-II stimulation. Both L-amlodipine and fluvastatin were beneficial to improve these adverse changes, and these favorable effects were further enhanced by L-amlodipine combined fluvastatin treatment. HUVECs apoptosis induced by Ang-II were mitigated by both L-amlodipine and fluvastatin, and this benefit was further promoted by L-amlodipine combined fluvastatin treatment. Notably, all these improvements achieved by L-amlodipine combined fluvastatin treatment were comparable to those of ROCK1 inhibitor. Ang-II is a significant contributor to EndMT via increasing ROCK1 expression and L-amlodipine combined fluvastatin treatment confers additive effects on improving EndMT and endothelium apoptosis, which may be beneficial for preventing hypertension development.