ABSTRACT
<p><b>OBJECTIVE</b>To investigate the lipopolysaccharide (LPS) antagonizing biological activity of densefruit pattany root-bark extract (DPR-2) in vitro.</p><p><b>METHODS</b>The effect of DPR-2 in neutralizing LPS (0.1 microg/L) was detected by kinetic turbidimetric limulus test. The effect of different concentrations of DPR-2 (0,8.0,16.0,32.0,64.0 mg/L) on binding of FITC-conjugated LPS (FITC-LPS,100.0 microg/L) to murine RAW264.7 cells was analyzed with laser scanning confocal microscopy. The expression of TNF-alpha and IL-6 mRNA in RAW264.7 cells after exposure to LPS (100.0 microg/L) were determined by real-time RT-PCR.</p><p><b>RESULTS</b>DPR-2 could neutralize LPS (P < 0.05 or P < 0.01), and inhibit the binding of FITC-LPS to RAW264.7 cells in a dose-dependent manner when the concentration of DPR-2 was above 16.0 mg/L. Furthermore, DPR-2 could markedly inhibit the expression of TNF-alpha and IL-6 mRNA in LPS-stimulated murine RAW264.7 cells.</p><p><b>CONCLUSION</b>DPR-2 exhibit an anti-LPS effect in vitro, which may be related to its capacity to neutralize LPS and inhibit binding of LPS for its receptors.</p>
Subject(s)
Animals , Mice , Cell Line , Drugs, Chinese Herbal , Pharmacology , Endotoxins , In Vitro Techniques , Limulus Test , Lipopolysaccharides , Monocytes , Metabolism , Plant Extracts , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the expression and analysis of its activity of anti-bacterial peptide gloverin in COS-7 cells.</p><p><b>METHODS</b>The appearance frequency of all genetic codes in the cDNA sequence from the same species of protein Attacin A was analyzed, and its cDNA sequence was synthesized by PCR overlapping extension method in conjunction with the designation of the known protein sequence of gloverin. The genes were inserted into pCDSI, an eukaryotic vector, after being identified correctly. As a result, the vector pBZHG was constructed. Thereafter, the liposome FuGENE( trade mark ) 6 was employed as the vector, and the COS-7 cells were transfected with liposome pBZHG and blank vector pCDSI. The normal cells were taken as the control. The supernatant was collected for the detection of its bactericidal activity after 72 PBHs.</p><p><b>RESULTS</b>The gloverin cDNA sequence designed artificially was expressed in COS-7 cells. The supernatant of the cells transfected by pBZHG exhibited bactericidal activity to E. coli J5 when compared with that from normal cells and in cells transfected with blank vectors.</p><p><b>CONCLUSION</b>The designed cDNA sequence of gloverin was proved to be genuine, and it provided the basis for future study of its antibiotic and anti-endotoxin activities.</p>
Subject(s)
Animals , Anti-Bacterial Agents , Pharmacology , COS Cells , Chlorocebus aethiops , Proteins , Genetics , Pharmacology , Sequence Analysis, DNA , TransfectionABSTRACT
Objective To investigate the role of bacterial DNA in systemic inflammatory response syndrome (SIRS). Methods A total of 100 mice of Kunming species were divided into ten groups: E.coli DNA (30, 20, 10, 5 and 1 mg/kg ), 30 mg/kg of CT DNA, 60Co DNA, DNased DNA, organic residue of DNA extraction and sterile water control. The last two were pre-treated with D-galactoamine (600 mg/kg intra peritoneally). Animals were administratively injected via tail vein. General physical condition and the death rate of mice were observed within 48 h. Results ①Obvious lethal effect of double strand E.coli DNA on mice were observed with a dose-effect correlation, LD50=11.51 mg/kg. ②NO difference in death rate was found in the group of 30 mg/kg E.coli DNA with or without 60Co irradiation (10/10 and 8/10,P>0.05). ③No rats died in the group of DNased DNA, organic residue of DNA extraction and calf thymic DNA (0/10). Conclusion Bacterial DNA may play an important role in the development of SIRS.
ABSTRACT
Objective To investigate the role of bacterial DNA in systemic inflammatory response syndrome (SIRS). Methods A total of 100 mice of Kunming species were divided into ten groups: E.coli DNA (30, 20, 10, 5 and 1 mg/kg ), 30 mg/kg of CT DNA, 60Co DNA, DNased DNA, organic residue of DNA extraction and sterile water control. The last two were pre-treated with D-galactoamine (600 mg/kg intra peritoneally). Animals were administratively injected via tail vein. General physical condition and the death rate of mice were observed within 48 h. Results ①Obvious lethal effect of double strand E.coli DNA on mice were observed with a dose-effect correlation, LD50=11.51 mg/kg. ②NO difference in death rate was found in the group of 30 mg/kg E.coli DNA with or without 60Co irradiation (10/10 and 8/10,P>0.05). ③No rats died in the group of DNased DNA, organic residue of DNA extraction and calf thymic DNA (0/10). Conclusion Bacterial DNA may play an important role in the development of SIRS.