Journal: Infection and Immunity

Article Title: Annexin A1 Attenuates Neutrophil Migration and IL-6 Expression through Fpr2 in a Mouse Model of Streptococcus suis -Induced Meningitis

doi: 10.1128/IAI.00680-20

Figure Lengend Snippet: The IL-6-inhibitory function of AnxA1 was mediated by the Fpr2/p38/COX-2 pathway. (A) BMDMs from wild-type (WT) and Fpr2 −/− mice were infected with S. suis (initial MOI of 100), and the supernatants of the cell cultures were collected to measure IL-6 levels via ELISA at the indicated time points. (B) BMDMs were treated with AnxA1, SB203580, nimesulide, anti-AnxA1 antibody, or isotype antibody before S. suis (initial MOI of 100) infection, and IL-6 levels were measured at 8 h. (C) The cellular proteins were subjected to Western blotting to assess COX-2, p-p38, p38, and GADPH expression. (D) Quantitative analyses of COX-2 and p-p38 expression. #/*, P < 0.05, ##/**, P < 0.01, and ***, P < 0.001, compared with the S. suis -infected group (#) in the same mouse strain or as indicated (*).

Article Snippet: In a separate experiment, WT and Fpr2 −/− BMDMs were pretreated with AnxA1 protein, 5 μM SB203580 (an inhibitor of p38 MAPK; MedChemExpress), 10 nM nimesulide (an inhibitor of COX-2; MedChemExpress), anti-AnxA1 antibody (1:200), or isotype control before S. suis infection (initial MOI of 100).

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing

Journal: Oncogene

Article Title: Celecoxib upregulates endoplasmic reticulum chaperones that inhibit celecoxib-induced apoptosis in human gastric cells.

doi: 10.1038/sj.onc.1209139

Figure Lengend Snippet: Figure 6 Changes in intracellular Ca2 þ concentration in the NSAID-dependent upregulation of GRP78. The intracellular Ca2 þ concentration was monitored by a fluo-3/AM assay system. Indicated concentrations of celecoxib were added to fluo-3/AM- loaded cells and the time–course of fluo-3 fluorescence change monitored. The maximum value for the increase in the intracellular Ca2 þ level (D[Ca2 þ]i) is shown (a). AGS cells were preincubated with or without 2 mM BAPTA-AM for 1 h and further incubated with or without 80 mM celecoxib (b–d), 800 mM nimesulide (e), 800 mM diclofenac (f) or 400 mM indomethacin (g) in the presence or absence of 2 mM BAPTA-AM for 6 h (celecoxib) or 12 h (other NSAIDs). The levels of GRP78 protein (b, e–g), GRP78mRNA (c) and ATF4 mRNA (d) were estimated by immunoblotting or real- time RT–PCR experiments as described in the legends of Figures 1 and 2. AGS cells were incubated with 100 mM celecoxib or 2 mM thapsigargin for indicated periods (h). Whole cell extracts (25 mg protein for ATF6 and 10 mg protein for actin) (upper panel in (h)) or nuclear extracts (20 mg protein for p50 ATF6 and 5 mg protein for lamin B) (lower panel in (h)) were analysed by immunoblotting with an antibody against ATF6, actin or lamin B as described in the legends of Figure 1. As for p50 ATF6 band, intensity of each band was expressed relative to the positive control sample (i.e. cells treated with thapsigargin for 1.5 h). Values shown are mean7s.d. (n ¼ 3). ***Po0.001; **Po0.01; *Po0.05.

Article Snippet: RPMI 1640 medium was obtained from Nissui Pharmaceutical Co. Fetal bovine serum (FBS) was obtained from Gibco Co. Pluronic F127, fluo-3/AM and BAPTA-AM were from Dojindo Co. Thapsigargin, staurosporine, diclofenac, RNaseA and propidium iodide (PI) were obtained from Sigma Co. PGE2 and indomethacin were obtained from Wako Co. Celecoxib was from LKT Laboratories Inc. Nimesulide was from Cayman Chemical Co. Antibodies against GRP78, ATF6, lamin, ATF4 and actin were purchased from Santa Cruz Biotechnology Inc., and those against phosphorylated PERK and phosphorylated eIF2a were from Cell Signaling Technology Inc.

Techniques: Concentration Assay, Incubation, Western Blot, Quantitative RT-PCR, Positive Control

Journal: Journal of Pain Research

Article Title: Effects of NSAIDs and paracetamol (acetaminophen) on protein kinase C epsilon translocation and on substance P synthesis and release in cultured sensory neurons

doi: 10.2147/JPR.S36916

Figure Lengend Snippet: Protein kinase C epsilon (PKCɛ) translocation induced by bradykinin (BK) and thrombin (THR) is inhibited by nimesulide and paracetamol, but not by other nonsteroidal anti-inflammatory drugs (NSAIDs) tested. ( A and B ) Confocal optical sections of cultured sensory neurons treated with THR (100 nM) for 30 seconds and subsequently fixed and stained for PCKɛ with a polyclonal specific antibody: ( A ) typical neuron showing no response; ( B ) typical THR-responsive neuron with translocation of PKCɛ to the plasma membrane. Neurons treated with 1 μM BK for 30 seconds had identical appearance. Neurons not treated with BK or THR showed no sign of translocation. Scale bar 5 μm. ( C ) PKCɛ translocation in cultured dorsal root ganglion neurons induced by THR (100 nM for 30 seconds), quantified as the percentage of neurons showing a clear edge. Translocation induced by THR was significantly reduced in the presence of nimesulide and paracetamol, while celecoxib, diclofenac, and ibuprofen were ineffective. All drugs were applied for 2 hours at a concentration of 10 μM before treatment with THR. ( D ) The effects of NSAIDs and paracetamol on translocation induced by BK (1 μM for 30 seconds) were largely similar and nimesulide was significantly stronger than paracetamol ( P < 0.05). Notes: Values represent means ± standard error of the mean of data from six to eight separate cultures. * P < 0.05 versus controls (analysis of variance followed by Bonferroni’s t -test).

Article Snippet: The following anti-inflammatory/analgesic drugs were used in this study: nimesulide (Helsinn Healthcare, Lugano, Switzerland) and ibuprofen, celecoxib, diclofenac, and paracetamol (all obtained from Sigma-Aldrich, Milan, Italy).

Techniques: Translocation Assay, Cell Culture, Staining, Concentration Assay

Journal: Journal of Pain Research

Article Title: Effects of NSAIDs and paracetamol (acetaminophen) on protein kinase C epsilon translocation and on substance P synthesis and release in cultured sensory neurons

doi: 10.2147/JPR.S36916

Figure Lengend Snippet: Modulation of basal and induced preprotachykinin (PPT) mRNA expression in cultured rat dorsal root ganglion cells by nonsteroidal anti-inflammatory drugs (NSAIDs) and paracetamol. ( A ) Basal expression of PPT mRNA was significantly reduced by nimesulide but was unchanged by treatment with other NSAIDs or paracetamol (all at 10 μM). ( B ) Nimesulide, paracetamol, celecoxib, and diclofenac significantly decreased upregulation of PPT mRNA expression induced by the inflammatory soup, while ibuprofen was ineffective. Notes: The amount of PPT is normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) levels by subtracting the average GAPDH CT value from the average PPT CT value then applying the comparative method (2 −ΔΔCT ) using untreated cells as calibrator (controls). All drugs were applied at 10 μM for 36 hours. Values represent means ± standard error of the mean of four to six experiments. * P < 0.05 versus respective controls (analysis of variance followed by Bonferroni’s t -test).

Article Snippet: The following anti-inflammatory/analgesic drugs were used in this study: nimesulide (Helsinn Healthcare, Lugano, Switzerland) and ibuprofen, celecoxib, diclofenac, and paracetamol (all obtained from Sigma-Aldrich, Milan, Italy).

Techniques: Expressing, Cell Culture

Journal: Journal of Pain Research

Article Title: Effects of NSAIDs and paracetamol (acetaminophen) on protein kinase C epsilon translocation and on substance P synthesis and release in cultured sensory neurons

doi: 10.2147/JPR.S36916

Figure Lengend Snippet: Modulation by nonsteroidal anti-inflammatory drugs and paracetamol of substance P (SP) release from dorsal root ganglion neurons after 70 minutes ( A ) and 36 hours ( B ) of treatment with inflammatory soup. Notes: At 70 minutes, only nimesulide significantly reduced SP release, while over longer times celecoxib and diclofenac were also effective. Values are means ± standard error of the mean of four to six experiments and are expressed as % of control cultures. * P < 0.05 versus respective inflammatory soup-stimulated cultures (analysis of variance followed by Bonferroni’s t -test).

Article Snippet: The following anti-inflammatory/analgesic drugs were used in this study: nimesulide (Helsinn Healthcare, Lugano, Switzerland) and ibuprofen, celecoxib, diclofenac, and paracetamol (all obtained from Sigma-Aldrich, Milan, Italy).

Techniques: