Inhibition of angiotension II type 1 receptor reduced human endothelial inflammation induced by low shear stress

Low shear stress induces endothelial cell apoptosis and monocyte adhesion by upregulating PECAM‑1 expression

“…The sirna sequences used in the present study were as follows: PECAM-1 forward, 5'-AUU CUG GUC UCG AGA AUU CUU-3' and reverse, 5'-GAA UUC UCG AGA CCA GAA UUU-3'; and PecaM-1 negative control (nc) forward, 5'-acG uGa CAC GUU CGG AGA ATT-3' and reverse, 5'-UUC UCC GAA CGU GUC ACG UTT-3' (Shanghai GenePharma Co., Ltd.). The lipofectamine ® -rnaiMaX transfection reagent (invitrogen; Thermo Fisher Scientific, inc) was utilized to transfect the HUVECs with PECAM-1 or NC siRNA as previously described (15). Cells at 70-80% confluency were co-cultured with 50 nmol/l PECAM-1 or 50 nmol/l NC siRNA and lipofectamine ® -RNAiMAX at 37˚C for 6 h. At 48 h post-transfection, the cells were either exposed to shear stress or not.…”

“…Shear stress experiment. Shear stress of 2 dyn/cm 2 was applied in vitro using a parallel-flow chamber as previously described (15,22,23). In brief, endothelial monolayers containing cell culture slides were subjected to lSS for 0, 30, 60 and 120 min after being placed in a parallel-plate flow chamber.…”

“…Western blotting analysis. Total protein of the HUVECs was obtained as previously described (15). The protein expression of PecaM-1, Foxo1, phospho-Foxo1, akt, and phospho-Akt in HUVECs were analyzed using western blotting.…”

“…increasing evidence has demonstrated that the endothelial dysfunction process is directly influenced by various factors, including nitric oxide dysfunction, epigenetic factors, oxidative stress, inflammatory factors and low shear stress (LSS) (5)(6)(7)(8)(9)(10)(11). among these, lSS, tangential stress produced by endothelial surface blood flow friction, which is observed at bifurcations, branching points and inside areas of curved segments of the coronary artery, induces inflammation and apoptosis, and subsequently disrupts the endothelial barrier, eventually leading to atherosclerosis (11)(12)(13)(14)(15). However, to date, no molecular targets have clearly linked lSS with the endothelial barrier.…”

No answer to the lack of specificity: mouse monoclonal antibody targeting the angiotensin II type 1 receptor AT1 fails to recognize its target

“…Polyclonal antibodies targeting both angiotensin II (AngII) receptors type 1 (AT1) and type 2 (AT2) have been described as non-specific (Benicky et al 2012;Herrera et al 2013;Hafko et al 2013). Despite these findings, antibodies directed against AT1 receptors are still widely used in the literature with more than twenty publications in 2017 using the Santa Cruz aspecific sc-1173 polyclonal antibodies (Chao et al 2017;Young et al 2017;Liu et al 2017) and already five publications in 2018 (Ahad et al 2018;Ishikane et al 2018;Rizzetti et al 2018;Zhao et al 2018;Ribeiro et al 2018).…”