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Objective To identify the activity of HCV IRES translation differences and identify the relationship between HCV IRES translation activity and ROS in different concentrations of ferric ammonium citrate ( FAC) in-duction.Methods 1 ) Expression plasmid pCI-Rluc-HCV IRES-Fluc was confirmed by endonuclease digestion as well as luciferase transient expression in Huh-7 cell;2) Controlled by dual-luciferase reporter assay, the differ-ent translation activity of HCV internal ribosomal entry site ( IRES ) was examined in a concentration of 50 μmol/L and 300μmol/L of FAC induction;ROS fluorescent staining method was used to detect the activity of ROS in Huh-7 cells, Western blot method was used to detect the protein expression changes of Nrf2 in Huh-7 cells;3) On the basis of the above experiments, 100 μmol/L DPI was added in 300 μmol/L FAC experimental group, to analyse the changes of HCV replication and ROS production after joining DPI.Results The generation of ROS and the activity of luciferase in the model group were significantly higher than that in the control group ( P<0.05 ) .FAC can enhance the expression of HCV IRES and increase the production of ROS , then causing Nrf2 expression in Huh-7 cell.However,after adding ROS inhibitor DPI, the above functions in Huh-7 cell were weakened.Conclusions The increase of HCV IRES expression induced by FAC is related to excessive ROS pro-duction induced by FAC in Huh-7 cells.
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Objective To investigate the role of heme oxygenase / carbon monoxide(HO /CO) pathway in the development of renal hypertension.Methods(1)The animal model of two-kidney-one clip renal hypertensive rats were prepared and the plasma contents of angiotensin Ⅱ(AngⅡ) were determined by radiommunoassay.(2)The activity of HO was tested at 2,4,6,8 weeks after surgery.(3) Western blot was used to estimated the level of HO-1 expression of aorta.Results In 2K1C rats,there was a significant increase in blood pressure at 2K1C.At the same time,rennin-agiotensin system(RAS) in plasma was activated and the content of AngⅡ obviously increased.The activity of HO was greater at 4-and 6-week in 2K1C group than sham group(P
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AIM:To investigate the role of heme oxygenase (HO) in AngⅡ induced proliferation and hypertrophy of cultured vascular smooth muscle cells. METHODS:(1) Western blotting analysis was carried out to detect protein level of HO-1 in the tissues. (2) [3H]-TdR,[3H]-leucine incorporation was measured in cultured vascular smooth muscle cells. (3) 2',7'-dichlorofluorescin diacetate (DCFH-DA) as an index was used to determine the cellular reactive oxygen species (ROS) level. RESULTS:(1) No significant difference in HO-1 protein expression level between AngⅡ-stimulated and control groups was observed,but HO-1 protein level in Hemin-induced group was higher than that in other two groups (P
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AIM: To investigate the effect of heme oxygenase on vascular remodeling in renal hypertension. METHODS: Male Wistar rats were randomly divided into sham-operated, 2K1C (two-kidney one-clip) and hemin-induced groups. Four weeks after the treatments, the thickness of aortic media and HO enzymatic activity of the aorta were determined. Immunohistochemical staining was carried out to detect protein of HO-1 in the aorta. RESULTS: The blood pressure in 2K1C renal hypertension rats started to increase two weeks after the surgery and stabled at a high level at the 4th week. Hemin, an inducer of HO-1, markedly inhibited the increase in blood pressure. Aortic medium thickness of the 2K1C rats at 4th week was 27 5% thicker than that in the sham-operated rats. The thickness of aortic medium of the hemin-induced rats was 16 1% less than that in 2K1C group. At the 4th week after operation, protein level and enzymatic activity of HO-1 in aorta were higher than that in 2K1C group compared to those in the sham-operated group. CONCLUSION: Renal hypertension caused vascular remodeling and the activation of HO-1. HO-1 induction decreased the blood pressure of renal hypertension and reduced vascular remodeling.
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AIM: To investigate the effect of heme oxygenase /carbon monoxide(HO/CO) system on the pathogenesis of hypertension and effect of Heme-L- Lysinate(HLL), Zine protoporphyrin-IX(ZnPPIX) on vascular tension in a two kidney one-clip model. METHODS: The changes in mean arterial pressure and heart rate as well as phenylephrine(PHE)-induced contraction in isolated aortic rings of HLL, ZnPPIX- treated rats were examined after intraperitoneal administration of HLL and carbon monoxide( CO). RESULTS: (1) Injection of exogenous CO resulted in a significant decrease in mean arterial pressure in Sham and 2K1C groups(P