Figure 3
BSNXD regulates estrogen receptor β mediating nitric oxide production and NF-κB suppression in human umbilical vein endothelial cells (HUVECs). The primary HUVECs were exposed to control serum or 10% BSNXD-derived serum for 48 h; in the final 24 h culture the ox-LDL was added. The supernatants were collected, and ERα/ERβ mRNA (a) and protein (b) expression were analyzed; eNOS expression (c), NO (d) and malondialdehyde (MDA) (e) concentrations were determined; eNOS expression, NO production was enhanced and MDA was downregulated by the drug-derived serum. The primary HUVECs were collected, dual-stained with Annexin V-FITC/PI, and then analyzed by FCM (f); the 10% drug-derived serum could inhibit the ox-LDL-induced apoptosis. BSNXD-derived serum inhibited the LPS-induced NF-κB transcription in HUVECs (g). BSNXD treatment increased ERβ transcription and translation in HUVECs. Pretreatment of selective ERβ antagonist (R, R-tetrahydrochrysene, R, RTHC) but not ERα antagonist (methyl-piperidino pyrazole, MPP) could block these effects on (eNOS, NO, MDA expression, apoptosis, NF-κB transcription, etc.), which is induced by the drug-derived serum. Meanwhile, pretreatment with the NO synthase inhibitor (NG-nitro-L-arginine methyl ester, L-NAME) significantly decreased the NO production, blocked the anti-apoptosis and NF-κB activity inhibition effect of BSNXD in HUVECs. Data are expressed as mean values±S.E.M. (n=6). *P<0.05, **P<0.01
