In-phase and out-of-phase voltages between electrodes were monitored and converted to TEER. Baseline resistance for each cell preparation was measured immediately before experimental manipulation. Statistical analysis Results are presented as mean �� standard deviation. Statistical significance was assessed by non-parametric Kruskal-Wallis one-way analysis of variance or a Student's t-test. For statistical comparisons of clinical values prior and subsequent to Olaparib clinical trial losartan treatment, unpaired t-tests were performed. P<0.05 was considered to indicate a statistically significant difference. Results AGEs upregulate ANGPTL4 mRNA and protein levels AGEs at 0, 20, 40, 80 or 160 ?g/ml<br>
were incubated with endothelial cells for 24 h. ANGPTL4
mRNA and protein levels were assayed by qPCR and western blot analysis. AGEs at 80 ?g/ml increased ANGPTL4 mRNA and protein levels significantly (2.60��0.18 vs. 1.00, 250��32 vs. 100%, respectively; P<0.05). BSA had no effect on ANGPTL4 levels in endothelial cells (P>0.05) (Fig. 1). Figure 1. Advanced glycation end products (AGEs) upregulate angiopoietin-like protein 4 (ANGPTL4) (A) mRNA and protein. (B) Western blots. Con, control; A20, 20 ?g/ml AGEs; A40, 40 ?g/ml AGEs; A80, 80 ?g/ml AGEs; A160, 160 ?g/ml ... AGEs increase Ang II levels in media and endothelial cell lysates AGEs at 0, 20, 40, 80 or 160 ?g/ml were incubated with endothelial cells for 24 h. Ang II levels in media Sclareol and lysates were Vemurafenib research buy measured by ELISA. AGEs dose-dependently increased Ang II levels in media and lysates. AGEs at 80 ?g/ml had the largest effect. AGEs at 160 ?g/ml did not have additional effects, although the effects remained significant (P<0.05). BSA had no effect on Ang II levels (P>0.05) (Table I). Table I. Advanced glycation end products (AGEs) increased angiotensin I levels in conditioned media and cell lysates. Losartan alleviates the upregulation
of ANGPTL4 and the increased endothelial permeability induced by AGEs To investigate the effects of losartan on induction of ANGPTL4 expression, AGEs were used to upregulate the expression of ANGPTL4 mRNA and protein. Pretreatment with losartan prevented upregulation of ANGPTL4 mRNA and protein induced by AGEs (1.52��0.21 vs. 2.60��0.18; 145��19 vs. 250��32%, respectively; P<0.05) (Fig. 2). AGEs at 80 ?g/ml increased endothelial permeability significantly and this increase was prevented by pretreatment with losartan (P<0.05) (Table II). Figure 2. Losartan inhibits upregulation of angiopoietin-like protein 4 (ANGPTL4) (A) mRNA and protein induced by advanced glycation end products (AGEs). (B) Western blots. Con, control; A20, 20 ?g/ml AGEs; A40, 40 ?g/ml AGEs; A80, 80 ?g/ml ... Table II. Fluorescein isothiocyanate-labeled dextran filtration assays and transendothelial electrical resistance (TEER).</div>
were incubated with endothelial cells for 24 h. ANGPTL4
mRNA and protein levels were assayed by qPCR and western blot analysis. AGEs at 80 ?g/ml increased ANGPTL4 mRNA and protein levels significantly (2.60��0.18 vs. 1.00, 250��32 vs. 100%, respectively; P<0.05). BSA had no effect on ANGPTL4 levels in endothelial cells (P>0.05) (Fig. 1). Figure 1. Advanced glycation end products (AGEs) upregulate angiopoietin-like protein 4 (ANGPTL4) (A) mRNA and protein. (B) Western blots. Con, control; A20, 20 ?g/ml AGEs; A40, 40 ?g/ml AGEs; A80, 80 ?g/ml AGEs; A160, 160 ?g/ml ... AGEs increase Ang II levels in media and endothelial cell lysates AGEs at 0, 20, 40, 80 or 160 ?g/ml were incubated with endothelial cells for 24 h. Ang II levels in media Sclareol and lysates were Vemurafenib research buy measured by ELISA. AGEs dose-dependently increased Ang II levels in media and lysates. AGEs at 80 ?g/ml had the largest effect. AGEs at 160 ?g/ml did not have additional effects, although the effects remained significant (P<0.05). BSA had no effect on Ang II levels (P>0.05) (Table I). Table I. Advanced glycation end products (AGEs) increased angiotensin I levels in conditioned media and cell lysates. Losartan alleviates the upregulation
of ANGPTL4 and the increased endothelial permeability induced by AGEs To investigate the effects of losartan on induction of ANGPTL4 expression, AGEs were used to upregulate the expression of ANGPTL4 mRNA and protein. Pretreatment with losartan prevented upregulation of ANGPTL4 mRNA and protein induced by AGEs (1.52��0.21 vs. 2.60��0.18; 145��19 vs. 250��32%, respectively; P<0.05) (Fig. 2). AGEs at 80 ?g/ml increased endothelial permeability significantly and this increase was prevented by pretreatment with losartan (P<0.05) (Table II). Figure 2. Losartan inhibits upregulation of angiopoietin-like protein 4 (ANGPTL4) (A) mRNA and protein induced by advanced glycation end products (AGEs). (B) Western blots. Con, control; A20, 20 ?g/ml AGEs; A40, 40 ?g/ml AGEs; A80, 80 ?g/ml ... Table II. Fluorescein isothiocyanate-labeled dextran filtration assays and transendothelial electrical resistance (TEER).</div>