par2  (Tocris)

Journal: Applied Sciences

Article Title: Trypsin Depolarizes Pacemaker Potentials in Murine Small Intestinal Interstitial Cells of Cajal

doi: 10.3390/app12094755

Figure Lengend Snippet: Figure 2. Effects of AC55541 on the pacemaker potentials in murine small intestinal interstitial cells of Cajal. (A–C) AC55541 (10–50 µM) dose-dependently depolarized pacemaker potentials and decreased their amplitudes. Responses to AC55541 are summarized in (D,E). Bars represent mean ± standard of mean. ** p < 0.01 compared with the control. CTRL, control.

Article Snippet: AC55541 (selective PAR2 agonist) [24], SR-140333 (NK1 receptor antagonist) [25], staurosporine (a broad-spectrum protein kinase C (PKC) inhibitor) [26], Go6976 (a calciumdependent PKC α/β inhibitor) [27], rottlerin (a calcium-independent PKC δ inhibitor) [28], HC067047 (selective TRPV4 antagonist) [29], and thapsigargin (inhibitor of Ca2+-ATPase in the endoplasmic reticulum) [30] were purchased from Tocris Bioscience (Bristol, UK).

Techniques: Control

Journal: Journal of Neuroinflammation

Article Title: Inhibition of mast cell tryptase attenuates neuroinflammation via PAR-2/p38/NFκB pathway following asphyxial cardiac arrest in rats

doi: 10.1186/s12974-020-01808-2

Figure Lengend Snippet: The schematic for the proposed mechanism underlying the anti-neuroinflammatory effect of MC-tryptase inhibition. We propose that following ACA, MCs will be activated and degranulated leading to the release of MC-derived tryptase in the brain, which will activate microglial PAR-2. Phosphorylation and activation of p38 and activation of NFκB in response to activated PAR-2 will result in the release of microglia-derived proinflammatory cytokines, IL-6, and TNF-α. Resultantly, neuroinflammation will contribute to neurocognitive dysfunction following ACA. For this study, we used selective MC-tryptase inhibitor APC366 for treatment purposes while PAR-2 activator AC55541 and p-38 inhibitor SB203580 were used for intervention

Article Snippet: AC55541 (selective PAR-2 activator; 30 μg/rat) and SB203580 (selective p38 inhibitor; 300 μg/rat; Santa Cruz Biotechnology, Dallas, TX, USA) were used for intervention.

Techniques: Inhibition, Derivative Assay, Phospho-proteomics, Activation Assay

Journal: Journal of Neuroinflammation

Article Title: Inhibition of mast cell tryptase attenuates neuroinflammation via PAR-2/p38/NFκB pathway following asphyxial cardiac arrest in rats

doi: 10.1186/s12974-020-01808-2

Figure Lengend Snippet: The number and distribution of the animals included for the present study

Article Snippet: AC55541 (selective PAR-2 activator; 30 μg/rat) and SB203580 (selective p38 inhibitor; 300 μg/rat; Santa Cruz Biotechnology, Dallas, TX, USA) were used for intervention.

Techniques: Staining, Western Blot, Mass Spectrometry

Journal: Journal of Neuroinflammation

Article Title: Inhibition of mast cell tryptase attenuates neuroinflammation via PAR-2/p38/NFκB pathway following asphyxial cardiac arrest in rats

doi: 10.1186/s12974-020-01808-2

Figure Lengend Snippet: APC366 improved short-term neurological deficits after ACA. a Effect of APC366 on neurological deficit score (NDS) at 24, 48, and 72 h and at 7 days after ACA. b Seizure activity at 24 and 48 h after ACA. c Adhesive tape removal at 48 and 72 h after ACA. d T-maze test at 7 days after ACA. ACA was associated with significantly worse neurologic functions compared to the shams in all tests except 7-day NDS. APC366 treatment at both doses of 50 μg and 150 μg improved NDS at 24, 48, and 72 h after ACA compared to the vehicle-treated ACA rats. APC366 at dose of 50 μg significantly reduced the seizure activity and improved performance of adhesive tape removal and T-maze tests compared to the ACA + vehicle group. PAR-2 activation with AC55541 exacerbated ACA-induced neurobehavioral deficits in all neurobehavioral tests except for the seizure activity at 24 h and adhesive tape removal at 72 h after ACA. Data are expressed as mean ± SD. n = 6/group. ANOVA, Tukey. ** p < 0.001 compared to sham, * p < 0.05 compared to sham, && p < 0.001 compared to ACA + vehicle, & p < 0.05 compared to ACA + vehicle

Article Snippet: AC55541 (selective PAR-2 activator; 30 μg/rat) and SB203580 (selective p38 inhibitor; 300 μg/rat; Santa Cruz Biotechnology, Dallas, TX, USA) were used for intervention.

Techniques: Activity Assay, Adhesive, Activation Assay

Journal: Journal of Neuroinflammation

Article Title: Inhibition of mast cell tryptase attenuates neuroinflammation via PAR-2/p38/NFκB pathway following asphyxial cardiac arrest in rats

doi: 10.1186/s12974-020-01808-2

Figure Lengend Snippet: APC366 reduced FJC-positive degenerating neurons at 7 days after ACA. a Representative FJC staining microphotographs. b Quantitative analyses of FJC-positive cells in (a) hippocampal subiculum, (b) CA1, and (c) CA2/3. Scale bar = 50 μm. ACA caused significant neuronal degeneration in the subiculum and CA1 regions. The activation of PAR-2 with AC55541 further exacerbated neuronal degeneration in rats subjected to ACA. APC366 at doses of 50 μg and 150 μg significantly reduced the number of FJC-positive cells in the subiculum and CA1 regions compared to the vehicle or AC55541-treated ACA rats. The tendency toward better treatment efficacy of 50 μg APC366 than 150 μg APC366 did not reach statistical significance. Data are expressed as mean ± SD. n = 6/group. ANOVA, Tukey. ** p < 0.001 compared to sham, * p < 0.05 compared to sham, && p < 0.001 compared to ACA + vehicle, & p < 0.05 compared to ACA + vehicle

Article Snippet: AC55541 (selective PAR-2 activator; 30 μg/rat) and SB203580 (selective p38 inhibitor; 300 μg/rat; Santa Cruz Biotechnology, Dallas, TX, USA) were used for intervention.

Techniques: Staining, Activation Assay

Journal: Journal of Neuroinflammation

Article Title: Inhibition of mast cell tryptase attenuates neuroinflammation via PAR-2/p38/NFκB pathway following asphyxial cardiac arrest in rats

doi: 10.1186/s12974-020-01808-2

Figure Lengend Snippet: PAR-2 activation reversed the anti-neuroinflammatory effects of APC366 at 24 h after ACA. Representative western blot images and quantitative analysis of MC-tryptase ( a ), PAR-2 ( b ), p38 ( c ), p-p38 ( d ), NFκB ( e ), and proinflammatory cytokines ( f , g ) in the brain at 24 h following ACA. The levels of the pathway proteins were markedly increased following ACA. The inhibition of MC-tryptase significantly reduced PAR-2, p-p38 and NFκB, TNF-α, and IL-6 levels in rats treated with APC366 compared to the ACA + vehicle group. Further activation of PAR-2 by AC55541 caused higher expressions of p-p38, NFκB, and proinflammatory cytokines compared to the vehicle-treated ACA group. The co-administration of APC366 and AC55541 not only abolished the protective effect of APC366 in ACA rats, but also offset the detrimental effects of pharmacological activation of PAR-2 by AC55541, resulting in significantly lower levels of p-p38, NFkB, IL-6, and TNF-a compared to AC55541 alone. Selective p38 inhibitor, SB203580 reversed the aggravated neuroinflammation due to AC55541 by decreasing the expressions of p-p38, NFκB, IL-6, and TNF-α. h Neurologic deficit score at 24 h following ACA. The inhibition of MC-tryptase significantly improved neurologic function. This effect was reversed with the activation of PAR-2 by AC55541 at 24 h following ACA. The activation of PAR-2 by AC55541 alone significantly worsened neurological performance; however, this effect was rescued by the p-38 inhibitor, SB203580, compared to the ACA + vehicle group. Data are expressed as mean ± SD. n = 6/group. ANOVA, Tukey. ** p < 0.001 compared to sham, * p < 0.05 compared to sham, && p < 0.001 compared to ACA + vehicle, & p < 0.05 compared to ACA + vehicle, $ p < 0.05 compared to ACA + APC366 (50 μg), % p < 0.05 compared to ACA + APC366 (50 μg) + AC55541 (30 μg), # p < 0.001 compared to ACA + AC55541 (30 μg), ## p < 0.05 compared to ACA + AC55541 (30 μg)

Article Snippet: AC55541 (selective PAR-2 activator; 30 μg/rat) and SB203580 (selective p38 inhibitor; 300 μg/rat; Santa Cruz Biotechnology, Dallas, TX, USA) were used for intervention.

Techniques: Activation Assay, Western Blot, Inhibition

Journal: Shock

Article Title: Inhibition of PAR-2 Attenuates Neuroinflammation and Improves Short-Term Neurocognitive Functions Via ERK1/2 Signaling Following Asphyxia-Induced Cardiac Arrest in Rats

doi: 10.1097/shk.0000000000001516

Figure Lengend Snippet: FIG. 3. Effect of FSLLRY-NH2 on neurocognitive functions after ACA. Effects of FSLLRY-NH2 on neurological functions were assessed by (A) NDS at 24 and 48 h, (B) number of seizures at 24 and 48 h, and (C) T-maze test at 7 days after ACA. ACA significantly worsened performance while treatment with FSLLRY- NH2 at a dose of 50 mg significantly improved neurological functions in all tests following ACA. FSLLRY-NH2 treatment at a dose of 100 mg was not effective in NDS at 24 h and T-maze test. AC55541 significantly deteriorated neurobehavioral outcome at all time points compared to the vehicle treated ACA group. Data are expressed as mean SD. n ¼ 6/group. ANOVA, Tukey. **P < 0.001 vs. Sham group, *P < 0.05 vs. Sham group, #P < 0.05 vs. ACA þ vehicle group. ACA indicates asphyxial cardiac arrest; d, days; h, hours.

Article Snippet: Selective PAR-2 activator AC55541 (30 mg/rat) and ERK1/2 inhibitor PD98059 (Santa Cruz Biotechnology, Dallas, Tex; 2 mL of 2 mmol/L) were used for intervention (25).

Techniques:

Journal: Shock

Article Title: Inhibition of PAR-2 Attenuates Neuroinflammation and Improves Short-Term Neurocognitive Functions Via ERK1/2 Signaling Following Asphyxia-Induced Cardiac Arrest in Rats

doi: 10.1097/shk.0000000000001516

Figure Lengend Snippet: FIG. 4. Effect of FSLLRY-NH2 on neuronal degeneration after ACA. Representative microphotographs (A) and quantitative analyses (B) of FJC-positive cells revealed significant neuronal degeneration in hippocampal CA1 region at 7 days after ACA. AC55541 further exacerbated the number of FJC-positive cells in rats subjected to ACA while FSLLRY-NH2 at doses of 50 mg and 150 mg markedly reduced hippocampal neuronal degeneration. Data are expressed as mean SD. n ¼ 6/group. ANOVA, Tukey. Scale bar ¼ 100 mm. **P < 0.001 vs. Sham group, ##P < 0.001 vs. ACA þ vehicle group, #P < 0.05 vs. ACA þ vehicle group. ACA indicates asphyxial cardiac arrest.

Article Snippet: Selective PAR-2 activator AC55541 (30 mg/rat) and ERK1/2 inhibitor PD98059 (Santa Cruz Biotechnology, Dallas, Tex; 2 mL of 2 mmol/L) were used for intervention (25).

Techniques:

Journal: Shock

Article Title: Inhibition of PAR-2 Attenuates Neuroinflammation and Improves Short-Term Neurocognitive Functions Via ERK1/2 Signaling Following Asphyxia-Induced Cardiac Arrest in Rats

doi: 10.1097/shk.0000000000001516

Figure Lengend Snippet: FIG. 5. Inhibition of ERK1/2 abolished the neuroinflammatory effect of PAR-2 activation at 24 h after ACA. Representative Western blot images (A) and quantitative analysis of PAR-2 (B), ERK1/2 (C), p-ERK1/2 (D), IL-6 (E), and TNF- a (F) in the brain revealed increased protein levels at 24 h following ACA except for ERK1/2 compared to the Sham group. Treatment with FSLLRY-NH2 significantly reduced p-ERK1/2 and proinflammatory cytokine levels compared to the ACA þ vehicle group. Further activation of PAR-2 with AC55541 only aggravated the neuroinflammatory response by increasing p-ERK1/2 expression. Potent ERK1/2 inhibitor PD98059 reversed the neuroinflammatory effect of AC55541. Neurologic outcome assessment with NDS at 24 h following ACA (G) revealed that the inhibition of PAR-2 significantly improved neurologic function while AC55541 alone significantly worsened performance compared to the ACA þ vehicle group. This detrimental effect of AC55541 on NDS was reversed by the ERK1/2 inhibitor, PD98059. Data are expressed as mean SD. n ¼ 6/group. ANOVA, Tukey. **P < 0.001 vs. Sham group, *P < 0.05 vs. Sham group, #P < 0.05 vs. ACA þvehicle group, &&P < 0.001 compared to ACA þ AC55541 group, &P < 0.05 compared to ACA þ AC55541 group. ACA indicates asphyxial cardiac arrest.

Article Snippet: Selective PAR-2 activator AC55541 (30 mg/rat) and ERK1/2 inhibitor PD98059 (Santa Cruz Biotechnology, Dallas, Tex; 2 mL of 2 mmol/L) were used for intervention (25).

Techniques: Inhibition, Activation Assay, Western Blot, Expressing