ABSTRACT
<p><b>OBJECTIVE</b>To explore the mechanism of tanshinol on alleviate the inflammatory injury of lung tissue in rat hepatopulmonary syndrome (HPS).</p><p><b>METHODS</b>SD rats were randomly divided into normal control group (n = 8), hepatopulmonary syndrome (HPS) group (n = 11) and tanshinol intervention group (n = 9). HE staining was used to observe the histopathology changes of pulmonary and hepatic tissues, and to count the number of macrophages in lung tissues. The activity of alanine transferase (ALT) and concentrations of endotoxin, tumor necrosis factor-a (TNF-alpha) and homocystein (Hcy) in plasma were detected. The concentrations of TNF-alpha, nitric oxide (NO) and malondialdehyde (MDA) and the activity of inducible nitric oxide synthase (iNOS) in the lung tissues were measured, respectively.</p><p><b>RESULTS</b>Thickened alveolar septum and increased macrophages were observed in lungs in HPS rat. After administered with tanshinol, the pulmonary pathological changes were alleviated and the number of macrophages in lung tissue was decreased compared with HPS group. The activity of ALT and the concentrations of endotoxin, TNF-alpha and Hcy in plasma ,and TNF-alpha, iNOS, NO and MDA in lung tissue in HPS group were higher than those of normal control group; meanwhile, those tanshinol group were less those that of HPS group.</p><p><b>CONCLUSION</b>Tanshinol may play an important role in delaying the development of HPS through protecting liver or directly antagonizing the effect of intestinal endotoxemia so as to alleviate the inflammatory reaction in lung tissue.</p>
Subject(s)
Animals , Male , Rats , Alanine Transaminase , Metabolism , Caffeic Acids , Pharmacology , Disease Models, Animal , Endotoxins , Blood , Hepatopulmonary Syndrome , Drug Therapy , Pathology , Homocysteine , Blood , Liver , Pathology , Lung , Pathology , Macrophages , Pathology , Malondialdehyde , Metabolism , Nitric Oxide , Metabolism , Nitric Oxide Synthase Type II , Metabolism , Rats, Sprague-Dawley , Tumor Necrosis Factor-alpha , BloodABSTRACT
<p><b>OBJECTIVE</b>To investigate HMGB-1 expression and its extracellular release of cultured primary hepatic parenchymal cells (HC) and Kupffer cells (KC) that were induced by lipopolysaccharides (LPS).</p><p><b>METHODS</b>Primary hepatic parenchymal cells and Kupffer cells were cultured in flasks, and some cells were treated with 500 microg/L LPS for 24 hours (induced group) and some were not treated with LPS and served as controls. All of the cells were repeatedly frozen-thawed, and the expression levels of HMGB1-mRNA and HMGB1 proteins were detected by semi-quantitative RT-PCR and Western blot respectively. Then HC and KC were subcultured in 24-well culture plates for 6 h, 12 h, 24 h and 48 h, and the HMGB1 protein in culture fluids was detected by Western blot at each time point.</p><p><b>RESULTS</b>Compared with the cells in the control group, the expression levels of HMGB1-mRNA in the induced group were significantly increased in both HC and KC at 24 h (t=31.32 and 45.90, P<0.05) and the protein levels of HMGB1 showed the same results (t=46.19 and 38.44, P<0.05). There was a small quantity of HMGB1 protein in the culture fluids of two control groups and the induced group of HC. However the HMGB1 protein in the induced group of KC were obviously increased with prolonged culture time (F=42.74, P<0.05). Compared with the control group, the level of HMGB1 protein in the induced group of KC was not increased at 6 h (t=9.57, P>0.05) but was significantly increased at 12 h, 24 h and 48 h (t=21.95, 32.39, 44.16, respectively P<0.05).</p><p><b>CONCLUSION</b>LPS could increase HMGB1 expression of HC and KC and HMGB1 release from KC, but not from HC. The results suggest that KC play an important role in triggering inflammation and liver injury.</p>
Subject(s)
Animals , Female , Rats , Cells, Cultured , HMGB1 Protein , Metabolism , Hepatocytes , Metabolism , Kupffer Cells , Metabolism , Lipopolysaccharides , Liver , Cell Biology , Metabolism , Pathology , RNA, Messenger , Genetics , Rats, WistarABSTRACT
<p><b>OBJECTIVES</b>To develop a new method for amplifying and sequencing the full-length of HBV genome.</p><p><b>METHODS</b>A pair of primers located at the nick region of HBV molecule and a thermostable polymerase with high fidelity and sensitivity were used. After cloning the PCR products into a plasmid, the sequences of HBV genome were analyzed.</p><p><b>RESULTS</b>The full-length of HBV genome were acquired using this method. The sensitivity and fidelity of the new method were also analyzed. The least quantity of initial templates was 10(2) and the artificial mutation rate was 1.2 bp/kb.</p><p><b>CONCLUSION</b>This method can be used in amplification and sequence analysis of the full-length of HBV genome on a large scale.</p>