ABSTRACT
Objective@#To investigate the effect and related mechanisms of RTA-408 on rat vascular smooth muscle cells (VSMCs) calcification induced by advanced glycation end products(AGE).@*Methods@#VSMCs were isolated from the aorta of Sprague Dawley rats and cultured in vitro. The fifth generation of VSMCs were randomly divided into 4 groups with random number table including control group(cells were incubated with normal medium for 2 days, then incubated with bovine serum albumin for 5 days),AGE group (cells were incubated with normal medium for 2 days, then incubated with 200 mg/L AGE for 5 days), experimental group(cells were incubated with 100 nmol/L RTA-408 for 2 days,then incubated with 200 mg/L AGE for 5 days),and RTA group(cells were incubated with 100 nmol/L RTA-408 for 2 days,then incubated with bovine serum albumin for 5 days). Cytosolic calciumin VSMC was measured using arsenazo Ⅲ assay. Von Kossa staining was utilized to detect the calcium deposition.The contents of malondialdehyde(MDA) and superoxide dismutase(SOD) in VSMCs were tested by appropriate kits.The protein expressions of osteopontin (OPN), alkaline phosphatase (ALP), nuclear factor E2 related factor 2(Nrf2), and NAD(P)H: quinone oxidoreductase 1(NQO1) were examined using Western blot.@*Results@#(1) Cytosolic calciumconcentration was significantly higher in AGE group than in control group((2.43±0.15) mmol/L vs. (1.23±0.09) mmol/L, P<0.01), which was significantly reduced in experimental group((1.62±0.18) mmol/L,P<0.01 vs. AGE group). (2) Calcium deposition in VSMCs was significantly upregulated in AGE group than in control group(3.64±0.50 vs. 1.00±0.12, P<0.01), and was downregulated in experimental group (1.56±0.37, P<0.01 vs. AGE group). (3) The MDA contents were higher((3.79±0.27) nmol/mg prot vs.(1.99±0.15) nmol/mg prot, P<0.01), while the SOD activities were lower((308.45±14.28) U/mg prot vs. (428.58±11.00) U/mg prot, P<0.01) in AGE group than in control group. The MDA contents were lower((2.37±0.19) nmol/mg prot vs. (3.79±0.27) nmol/mg prot, P<0.01),while the SOD activities were higher((391.03±22.92) U/mg prot vs. (308.45±14.28) U/mg prot, P<0.05)in experimental group compared with AGE group. (4) The relative expressions of OPN and ALP were higher in AGE group than in control group(3.06±0.21 vs. 1.00±0.07, and 2.89±0.29 vs. 1.00±0.10,both P<0.01), both (OPN(1.15±0.12) and ALP(1.45±0.15)) were downregulated in experimental group (both P<0.01 vs. AGE group). (5) The relative protein expressions of Nrf2 and NQO1 in experimental group were higher than AGE group(2.37±0.17 vs. 1.17±0.09, and 3.91±0.18 vs. 1.05±0.08, both P<0.01).@*Conclusion@#Activation of nrf2/NQO1 signaling pathway by RTA-408 can reduce the AGE-induced VSMC calcification through attenuating oxidative injury.
ABSTRACT
Objective To investigate the pathological and immunohistochemical characteristics of female bladder ectopic skene glands.Methods A female with bladder tumor was treated in our hospital in May 2013.Preoperative so(n)graphy revealed a 0.9 cm×0.6 cm round solid mass in the bottom of bladder wall.Mass was hypoechoic homogeneous with regular shape,blood flow within the mass was noted.The tumor was treated with transurethral resection.Routine pathological examination suggested bladder ectopic Skene glands.Immunohistochemical stains for prostate specific antigen (PSA),prostate spectific acid phosphatase (PSAP),androgen receptor (AR),estrogen receptor (ER),CD10,cytokeratin 14 (CK14),cytokeratin 18 (CK18),P63,high molecular weight cytokeratin (34βE12),α-methylacyl-CoA racemase (AMACR/p504s) were further performed.Results Routine pathological examination showed prostate glands composed of prostate gland epithelial cells and basal cells in a submucosal location.Immunohistochemical stains showed:PSA-,PSAP +,AR +,ER-,CD10+,CK18 +,CK14-,P63 +,34βE12 +,AMACR-.Conclusions Routine pathological examination combined with immunohistochemical stains such as PSA,PSAP,and others,can be used to diagnose ectopic Skene glands disease.Female bladder ectopic Skene glands is a benign lesion,and the prognosis is good.