Monocyte chemoattractant protein-1 and osteopontin expression in NZB/W mesangial cells with lipopolysaccharide treatment

<p><b>Copyright information:</b></p><p>Taken from "Mesangial cells of lupus-prone mice are sensitive to chemokine production"</p><p>http://arthritis-research.com/content/9/4/R67</p><p>Arthritis Research & Therapy 2007;9(4):R67-R67.</p><p>Published online 7 Jul 2007</p><p>PMCID:PMC2206365.</p><p></p> Growth-arrested (under 2% fetal bovine serum (FBS)) mesangial cells were incubated with 10 μg/ml lipopolysaccharide (LPS) for different periods of time, and then levels of monocyte chemoattractant protein-1 (MCP-1) mRNA or osteopontin (OPN) mRNA were examined by real-time PCR. MCP-1 protein levels in the culture supernatant were measured by ELISA. OPN protein levels in the cell lysate were measured by western blot analysis; semiquantitative data are shown. The MCP-1 protein expression levels at various time points were normalized to total protein (pg/mg). The experiment was performed in triplicate, and results are expressed as the mean ± standard error, = 6. The last time point without LPS stimulation (unstimulated) was presented as the negative control. *< 0.05, **< 0.01, ***< 0.005

NF-κB inhibitor effect on lipopolysaccharide-induced monocyte chemoattractant protein-1 and osteopontin expression in NZB/W mesangial cells

<p><b>Copyright information:</b></p><p>Taken from "Mesangial cells of lupus-prone mice are sensitive to chemokine production"</p><p>http://arthritis-research.com/content/9/4/R67</p><p>Arthritis Research & Therapy 2007;9(4):R67-R67.</p><p>Published online 7 Jul 2007</p><p>PMCID:PMC2206365.</p><p></p> Growth-arrested (under 2% fetal bovine serum (FBS)) mesangial cells were preincubated for 2 hours with -tosyl-1-phenylalanine chloromethyl ketone (TPCK) or dexamethasone (Dex), and then 10 μg/ml lipopolysaccharide (LPS) was added for 12 hours: monocyte chemoattractant protein-1 (MCP-1) mRNA levels and osteopontin (OPN) mRNA levels were measured by real-time PCR analysis. Growth-arrested (under 2% FBS) mesangial cells were preincubated for 2 hours with TPCK or Dex, then 10 μg/ml LPS was added for 24 hours: MCP-1 protein levels in the supernatant were measured by ELISA, and OPN protein levels in the cell lysate were measured by western blot analysis; semiquantitative data are shown. The MCP-1 protein expression levels at various time points were normalized to total protein (pg/mg). The experiment was performed in triplicate, and results are expressed as the mean ± standard error. **< 0.01, ***< 0.005, = 6. White bar, absence of LPS; black bar, LPS alone; hatched bar, TPCK and LPS or Dex and LPS

Toll-like receptor 4 and myeloid differentiation factor 88 mRNA in NZB/W mesangial cells (lipopolysaccharide treatment)

<p><b>Copyright information:</b></p><p>Taken from "Mesangial cells of lupus-prone mice are sensitive to chemokine production"</p><p>http://arthritis-research.com/content/9/4/R67</p><p>Arthritis Research & Therapy 2007;9(4):R67-R67.</p><p>Published online 7 Jul 2007</p><p>PMCID:PMC2206365.</p><p></p> Growth-arrested (under 2% fetal bovine serum (FBS)) mesangial cells were incubated with 10 μg/ml lipopolysaccharide (LPS) for different periods of time, and then Toll-like receptor 4 (TLR-4) and myeloid differentiation factor 88 (MyD88) mRNA levels were measured by real-time PCR analysis. The experiment was performed in triplicate, and results are expressed as the mean ± standard error, = 6. The last time point without LPS stimulation (unstimulated) was presented as the negative control. *< 0.05, **< 0.01

Effects of procainamide on creatinine in LPS-induced rhabdomyolysis rats.

<p>Depicted are the changes during the experimental period in different groups of animals that received vehicle at time 0 (Control, n = 6), received vehicle at time 0 and then received procainamide at 1 h (Control + Pro, n = 6), received LPS at time 0 (LPS-induced rhabdomyolysis, n = 8), and received LPS at time 0 and then received procainamide at 1 h (LPS-induced rhabdomyolysis + Pro, n = 8). All data are expressed as mean ± SEM. *<i>P</i> < 0.05, all versus Control rats; ^#<i>P</i> < 0.05, with versus without procainamide in LPS-induced rhabdomyolysis rats.</p

Effects of procainamide on plasma IL-6 levels in LPS-induced rhabdomyolysis rats.

<p>Effects of procainamide on plasma IL-6 levels in LPS-induced rhabdomyolysis rats.</p

Effects of procainamide on superoxide levels in kidneys obtained from LPS-induced rhabdomyolysis rats.

<p>Depicted are changes at the end of experiments (at 6 h) in different groups of animals that received vehicle at time 0 (Control, n = 6), received vehicle at time 0 and then received procainamide at 1 h (Control + Pro, n = 6), received LPS at time 0 (LPS-induced rhabdomyolysis, n = 6), and received LPS at time 0 and then received procainamide at 1 h (LPS-induced rhabdomyolysis + Pro, n = 6). All data are obtained using the same rats in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0150319#pone.0150319.g001" target="_blank">Fig 1</a> and expressed as mean ± SEM. *<i>P</i> < 0.05, all versus Control rats; ^#<i>P</i> < 0.05, with versus without procainamide in LPS-induced rhabdomyolysis rats.</p

(A) Representative histopathologic features and (B) polymorphonuclear neutrophil (PMN) infiltration index of kidney tissue sections obtained from LPS-induced rhabdomyolysis rats.

<p>Depicted are changes in PMN infiltration index at the end of experiment (at 6 h) in different groups of animals that received vehicle at time 0 (Control, n = 3), received vehicle at time 0 and then received procainamide at 1 h (Control + Pro, n = 3), received LPS at time 0 (LPS-induced rhabdomyolysis, n = 3), and received LPS at time 0 and then received procainamide at 1 h (LPS-induced rhabdomyolysis + Pro, n = 3). All data are obtained using the same rats in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0150319#pone.0150319.g001" target="_blank">Fig 1</a> and expressed as mean ± SEM. *<i>P</i> < 0.05, all versus Control rats; ^#<i>P</i> < 0.05, with versus without procainamide in LPS-induced rhabdomyolysis rats. Arrows represent neutrophil infiltration. Original magnification x 400.</p

Effects of procainamide on the expressions of (A) DNMT1, (B) DNMT3A, and (C) DNMT3B in the lung of LPS-induced rhabdomyolysis rats.

<p>Depicted are changes in lung DNMT1, DNMT3A, and DNMT3B expressions at the end of experiment (at 6 h) in different groups of rats that received vehicle at time 0 (Control, n = 3), received vehicle at time 0 and then received procainamide at 1 h (Control + Pro, n = 3), received LPS at time 0 (LPS-induced rhabdomyolysis, n = 3), and received LPS at time 0 and then received procainamide at 1 h (LPS-induced rhabdomyolysis + Pro, n = 3). All data are obtained using the same rats in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0150319#pone.0150319.g001" target="_blank">Fig 1</a> and expressed as mean ± SEM. *<i>P</i> < 0.05, all versus Control rats; ^#<i>P</i> < 0.05, with versus without procainamide in LPS-induced rhabdomyolysis rats.</p

NF-κB p65 activation in NZB/W mesangial cells with lipopolysaccharide treatment

<p><b>Copyright information:</b></p><p>Taken from "Mesangial cells of lupus-prone mice are sensitive to chemokine production"</p><p>http://arthritis-research.com/content/9/4/R67</p><p>Arthritis Research & Therapy 2007;9(4):R67-R67.</p><p>Published online 7 Jul 2007</p><p>PMCID:PMC2206365.</p><p></p> Growth-arrested (under 2% fetal bovine serum (FBS)) mesangial cells (MCs) were incubated with 10 μg/ml lipopolysaccharide (LPS) for different periods of time, and then the distribution of NF-κB p65 was examined. Immunofluorescence: images a–d, NZB/W MCs; images e–h, DBA/W MCs (nonimmune strain, served as control) incubated for 0–12 hours with LPS; and image i, semiquantitative data. Electrophoresis mobility-shift assay performed using a DIG-labeled synthetic oligonucleotide and nuclear extract from MCs. The competition assay used the same unlabeled oligonucleotide at a 10-fold higher concentration. Arrow, NF-κB p65 binding bands. Comp., the abbreviation of competition. ELISA performed using the TransAM NF-κB p65 kit. The NF-κB p65 expression levels at various time points were normalized to nuclear protein (ng/mg). The experiment was performed in triplicate, and results are expressed as the mean ± standard error, = 6. The last time point without LPS stimulation (unstimulated) was presented as the negative control. ***< 0.005 versus DBA/W MC controls

Effects of procainamide on (A) potassium and (B) calcium levels in the arterial blood of LPS-induced rhabdomyolysis rats.

<p>Depicted are the changes during the experimental period in different groups of animals that received vehicle at time 0 (Control, n = 5), received vehicle at time 0 and then received procainamide at 1 h (Control + Pro, n = 5), received LPS at time 0 (LPS-induced rhabdomyolysis, n = 6), and received LPS at time 0 and then received procainamide at 1 h (LPS-induced rhabdomyolysis + Pro, n = 6). The values of all groups are normalized at 0 h as zero. All data are obtained using the same rats in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0150319#pone.0150319.g001" target="_blank">Fig 1</a> and expressed as mean ± SEM. *<i>P</i> < 0.05, all versus Control rats; ^#<i>P</i> < 0.05, with versus without procainamide in LPS-induced rhabdomyolysis rats.</p