Journal: Shock (Augusta, Ga.)

Article Title: Evodiamine represses hypoxia-induced inflammatory proteins expression and hypoxia-inducible factor 1alpha accumulation in RAW264.7.

doi: 10.1097/SHK.0b013e31819940cb

Figure Lengend Snippet: FIG. 1. Effect of evodiamine on hypoxia-induced inflammatory response. RAW264.7 cells were incubated in the presence or absence of indicated concentration of evodiamine for 1 h and then incubated in normoxia (21% O2) or hypoxia (1% O2) for 24 h. A, Equal amount of total proteins was subjected to sodium dodecyl sulfateYpolyacrylamide gel electrophoresis to detect COX-2, iNOS, and actin expressions by Western blotting analysis. B, The culture medium was assayed by PGE2 ELISA kit. C, Cellular viability was assayed by SRB assay. N indicates normoxia; H, hypoxia, respectively. ***P G 0.001 compared with normoxic cells, and ##P G 0.01 compared with hypoxic cells. The representative result of three separate experiments is shown.

Article Snippet: Prostaglandin E2 (PGE2) assay kit was from R&D Systems (Minneapolis, Minn).

Techniques: Incubation, Concentration Assay, Nucleic Acid Electrophoresis, Western Blot, Enzyme-linked Immunosorbent Assay, Sulforhodamine B Assay

Journal: Nature Communications

Article Title: The endocannabinoid anandamide is an airway relaxant in health and disease

doi: 10.1038/s41467-022-34327-0

Figure Lengend Snippet: a Statistical analysis of airway tone in tracheal rings from C57BL/6J mice indicates that AEA (10 µM, n = 5)-dependent airway relaxation is mediated via arachidonic acid (AA) and COX-dependent metabolites; arachidonic acid (AA, 10 µM, n = 7), nordihydroguaiaretic acid (NDGA, 10 µM; n = 9), 17-octadecynoic acid (ODYA, 1 µM, n = 8), indomethacin (Indo, 10 µM, n = 8), and Indo+URB597 ( n = 8). Note that data for URB ( n = 6 ) are taken from Fig. . All results were derived from independent samples. One-way ANOVA, Tukey’s post hoc test (AEA vs URB ***p = 1.6 × 10 −6 ; AEA vs Indo+URB ***p = 4.4 × 10 −7 ; AEA vs Indo ***p = 2.3 × 10 −4 ; NDGA vs Indo ***p = 2.5 × 10 −4 ; ODYA vs Indo **p = 0.0014). b Statistical analysis of airway tone in tracheal rings from C57BL/6J mice demonstrates strong inhibition of AA-dependent relaxation by Indo (AA, n = 7; Indo n = 6, independent samples). Unpaired two-tailed Student’s t test ***p = 1.4 × 10 −6 . c Statistical analysis of airway tone in tracheal rings from C57BL/6J mice reveals that AEA-induced airway relaxation is dependent on EP2 and EP4 receptors, AEA ( n = 5 ), CAY10441 (CAY, 3 µM, n = 3), AH + L ( n = 4), AH6809 (AH, 10 µM, n = 4), and L161,982 (L, 2 µM, n = 4). Results were derived from independent samples. One way ANOVA, Tukey’s post hoc test (AEA vs AH **p = 0.0047; AEA vs AH + L ***p = 3.2 × 10 −5 ; CAY vs AH + L **p = 0.0014; AH + L vs L **p = 0.0035. d AEA evokes PGE2 production in the supernatant of trachea (C57BL/6J mice, n = 6 independent samples) in a time-dependent manner. Unpaired two-tailed t test with Welch correction **p = 0.0079. e AEA induces cAMP production in tracheal tissue from C57BL/6J mice ( n = 4 independent samples) in a time-dependent manner. Unpaired two-tailed Student’s t test *p = 0.022. f AEA increases PGE2 production in the supernatant of human tracheal epithelial cells (hTEPC) after 60 min ( n = 5 independent samples). Two way ANOVA, Bonferroni’s post hoc test *p = 0.037. Values are expressed as box and whiskers plot, with boxes indicating the Q1 and Q3 ranges, center line representing the median and whiskers as minimum and maximum values. g AEA increases PGE2 production in human airway smooth muscle cells (hASMC) after 30 and 60 min ( n = 5 independent samples). Two way ANOVA, Bonferroni’s post hoc test (30 min: **p = 3.5 × 10 −5 , 60 min: **p = 7.1 × 10 −5 . Values are expressed as box and whiskers plot, with boxes indicating the Q1 and Q3 ranges, center line representing the median and whiskers as minimum and maximum values. h AEA increases cAMP production in hASMC after 60 min ( n = 5 independent samples). Two way ANOVA, Bonferroni’s post hoc test *p = 0.012. Values are expressed as box and whiskers plot, with boxes indicating the Q1 and Q3 ranges, center line representing the median and whiskers as minimum and maximum values. i – l Statistical analysis of airway relaxation by isoprenaline (ISO, 10 µM) ( i , k ) or AEA (10 µM) ( j , l ) after long-term treatment of tracheal rings from C57BL/6J mice with medium (left bar, n = 7 ( i , j ), n = 8 ( k , l )), ISO (right bar, n = 8 ( i , j )) or AEA (right bar, n = 7 ( k , l )) reveals that prolonged ISO incubation reduces ISO-dependent but not AEA-dependent relaxation while AEA incubation has no effect. Unpaired two-tailed Student’s t test *p = 0.012 ( I ). a – e , i – l Data are presented as mean values ± SEM. Source data are provided as a Source Data file.

Article Snippet: PGE2 was quantified in the supernatants using the Prostaglandin E2 ELISA Kit – Monoclonal (514010, Cayman Chemical, USA) and cAMP concentrations of tissue or cell lysates were determined with the Direct cAMP ELISA Kit (ADI-900-006, Enzo Life Sciences, Germany) according to the manufacturer’s instructions.

Techniques: Derivative Assay, Inhibition, Two Tailed Test, Incubation

Journal: Frontiers in Immunology

Article Title: Polydatin Prevents Lipopolysaccharide (LPS)-Induced Parkinson's Disease via Regulation of the AKT/GSK3β-Nrf2/NF-κB Signaling Axis

doi: 10.3389/fimmu.2018.02527

Figure Lengend Snippet: PLD suppresses the phosphorylation of NF-κB p65 and the release of pro-inflammatory mediators in LPS-activated BV-2 cells via Nrf2 activation. Following pretreatment with BT (Brusatol: an inhibitor of Nrf2, 200 nM) for 6 h, BV-2 cells were treated with PLD for 1 h and then stimulated with LPS for 1 h. (A) After cells were harvested, the protein expression of p-NF-κB p65 and NF-κB p65 was measured via Western blotting. (B) Levels of NO in culture supernatants were measured using the Griess reagent. Levels of PGE2 (C) , TNF-α (D) , IL-6 (E) , and IL-1β (F) in culture supernatants were measured via ELISA. Similar results were obtained from three independent experiments. Values are presented as the mean ± SEM ( n = 4 in each group), ## p < 0.01, vs. control group; ** p < 0.01, vs. LPS group; $$ p < 0.01, vs. BT+PLD+LPS group.

Article Snippet: Rat or mouse PGE2, IL-6, IL-1β, and TNF-α ELISA kits were obtained from Biolegend (San Diego, CA, USA).

Techniques: Phospho-proteomics, Activation Assay, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Control