ABSTRACT
Objective To investigate the effect of syntenin-1 on the assembly of hepatitis C virus (HCV) particles. Methods The content of syntenin-1 in human primary hepatocytes was detected by Western blotting analysis. Human hepatocellular carcinoma cell line Huh7.5.1 was transfected with lentiviral plasmids to construct syntenin-1 overexpressed cell line and green fluorescent protein (GFP) overexpressed cell line. After transferring HCV RNA into Huh7.5.1 cells, syntenin-1 overexpressed cells and GFP overexpressed cells by electroporation, the effects of syntenin-1 overexpression on HCV in the cells were investigated in terms of RNA replication, viral protein contents and secretion of infectious virus particles by luciferase analysis, Western blotting and virus titer, respectively. The density of viral particles was assessed by isopycnic ultracentrifugation to analyze the distributions of HCV RNA and infective titer. Results The contents of syntenin-1 in three human primary hepatocytes were higher than that in the Huh7.5.1 cells (P0.01). Syntenin-1 overexpression had no effect on HCV RNA replication in host cells; the infectivity of HCV derived from syntenin-1 overexpressed cells was not significantly different compared with the Huh7.5.1 cells or GFP overexpressed cells. In syntenin-1 overexpressed cell culture supernatants, some infectious HCV particles mainly concentrated in the region of concentration of 1.08-1.16 g/mL gradually transferred to the low-density region of 1.01-1.02 g/mL. Conclusion Syntenin-1 overexpression alters the distribution of density components of infectious HCV particles.
ABSTRACT
OBJECTIVE@#To study the damage of DNA in lymphocytes, brain cells and cardiac muscle cells of rats induced by different dose of tetramine and to speculate the toxicant mechanism of tetramine.@*METHODS@#The rat were poisoned by Tetramine, which was taken in by mouth. The rat poisoning models were used by 0.2, 0.1, 0.05, 0.01 mg x kg(-1) Tetramine, and comparison model was made by NS. Lymphocytes and brain cells and cardiac muscle cells of rats were separatd and collected form experimentation rat. DNA damages of cells which were exposed to different doses of tetramine were detected using the single cell gel electrophorresis (SCGE) or comet assay.@*RESULTS@#DNA damages have been observed in lymphocytes, brain cells and cardiac muscle cells of rats which exposed form 0.01mg x kg(-1) doses of tetramine to 0.2mg x kg(-1) doses of tetramine. The test groups are very significantly statistical different to the control group (P<0.01).@*CONCLUSION@#It is assumed that DNA damages of cells might be one of the toxicant mechanism of tetramine.
Subject(s)
Animals , Rats , Brain/pathology , Bridged-Ring Compounds/poisoning , Comet Assay/methods , DNA Damage , Dose-Response Relationship, Drug , Electrophoresis, Agar Gel/methods , Lymphocytes/drug effects , Myocardium/pathology , Rats, Sprague-DawleyABSTRACT
OBJECTIVE@#This study was performed to detect the phosphorylation state of Cx43 in human left ventricular myocardium among sudden deaths caused by acute myocardial ischemia (AMI) especially sudden coronary death (SCD) and control groups. And then evaluate the significance of these findings in diagnosing the early pathological changes of acute myocardial ischemia.@*METHODS@#Immunohistochemistry (IHC) SP techniques were adopted to detect the phosphorylation state of Cx43 in the left ventricular myocardium samples of 45 deceased, which classified as group I--SCD, group II & III (other two groups of AMI) and Group IV & V (two control groups, sudden death caused by lethal acute cranio-cerebral injury or pathologic intracranial hemorrhage). In addition, we selected anti-Pan-Cadherin (construction protein of adherent junctions on the intercalated disc) and PHA-E+L/Bio, to detect the integration of myocardial mechanical coupling and membranes (applying affinityhistochemistry, AHC) respectively.@*RESULTS@#(1) Phosphorylated Cx43 positive staining was almost invisible in Group I, II and III or scattered in sarcoplasm in few samples; but it was assembling at the IDs clearly in group IV and V. (2) Strongly positive staining of Pan-Cadherin could be observed at the IDs and (3) integrated myocardial membranes were found in all samples.@*CONCLUSION@#These findings suggested that compared with the control groups, the integration of myocardial mechanical coupling and membranes did not alter in AMI. But Cx43, the key protein of electrical coupling on myocardial gap junctions, occurred dephosphorylation remarkably in AMI. Thus applying IHC techniques to detect the Cx43 dephosphorylation in human left ventricular myocardium maybe useful to recognize the onset of arrhythmia in AMI, especially in SCD whose myocardium without apparent morphological changes.