Journal: Journal of Cellular and Molecular Medicine

Article Title: The α2AR/Caveolin‐1/p38MAPK/NF‐κB axis explains dexmedetomidine protection against lung injury following intestinal ischaemia‐reperfusion

doi: 10.1111/jcmm.16614

Figure Lengend Snippet: Dex reduces lung injury following intestinal I/R via Cav‐1‐dependent p38MAPK/NF‐κB pathway inactivation. A, Comparison of the down‐regulation for Cav‐1 protein using three anti‐Cav‐1 shRNA constructs. * P < .05 compared with the mock group. Pre‐injections were applied four days before intestinal I/R injury. B, RT‐qPCR of Cav‐1 mRNA expression and Western blot analysis of Cav‐1, p38, p‐p38, p‐p65 and p65 expression in rat lung tissues with Cav‐1 overexpression or knockdown before intestinal I/R injury. C, ELISA detection of TNF‐α expression in rat lung tissues with Cav‐1 overexpression or knockdown before intestinal I/R injury. D, ELISA detection of IL‐1β expression in rat lung tissues with Cav‐1 overexpression or knockdown before intestinal I/R injury. E, ELISA detection of IL‐6 expression in rat lung tissues with Cav‐1 overexpression or knockdown before intestinal I/R injury. F, Lung injury score of rat lung tissues with Cav‐1 overexpression or knockdown before intestinal I/R injury. G, Apoptosis rate of rat lung tissues with Cav‐1 overexpression or knockdown before intestinal I/R injury. * P < .05 vs sham group, # P < .05 vs I/R + oe‐NC group. H, Western blot analysis of Cav‐1, p38, p‐p38, p‐p65 and p65 in rat lung tissues upon Dex treatment and/or Cav‐1 knockdown prior to intestinal I/R injury. I, ELISA detection of TNF‐α expression in rat lung tissues upon Dex treatment and/or Cav‐1 knockdown prior to intestinal I/R injury. J, ELISA detection of IL‐1βexpression in rat lung tissues upon Dex treatment and/or Cav‐1 knockdown prior to intestinal I/R injury. K, ELISA detection of IL‐6 expression in rat lung tissues upon Dex treatment and/or Cav‐1 knockdown prior to intestinal I/R injury. L, Lung injury score of rat lung tissues upon Dex treatment and/or Cav‐1 knockdown prior to intestinal I/R injury. M, Apoptosis rate of rat lung tissues upon Dex treatment and/or Cav‐1 knockdown prior to intestinal I/R injury, detected by TUNEL staining. * P < .05 vs sham group, # P < .05 vs I/R + Dex +sh‐NC group. Data among multiple groups were analysed by one‐way ANOVA with Tukey's post hoc test

Article Snippet: The PVDF membrane was blocked in 5% skimmed milk at room temperature for 1 hours and incubated overnight at 4°C with the diluted primary antibodies: rabbit anti‐α 2A ‐AR (1:1000, 14266‐1‐AP, Proteintech ProteinTech Group), rabbit anti‐Cav‐1 (1:1000, ab32577, Abcam Inc.), rabbit anti‐p38 (1:1000, ab31828, Abcam), rabbit anti‐phosphorylated (p)‐p38 (1:1000, ab4822, Abcam), rabbit anti‐NF‐κBp65 (1:1000, ab16502, Abcam), rabbit anti‐NF‐κB‐p‐p65 (1:1000, ab86299, Abcam) and rabbit anti‐Actin (1:1000, ab179467, Abcam).

Techniques: Comparison, shRNA, Construct, Quantitative RT-PCR, Expressing, Western Blot, Over Expression, Knockdown, Enzyme-linked Immunosorbent Assay, TUNEL Assay, Staining

Journal: Anesthesia and Analgesia

Article Title: Dexmedetomidine Pretreatment Protects Against Myocardial Ischemia/Reperfusion Injury by Activating STAT3 Signaling

doi: 10.1213/ANE.0000000000006487

Figure Lengend Snippet: Dexmedetomidine activates α2-AR to promote STAT3 phosphorylation. A, Volcano plot of 2257 differentially expressed mRNAs induced by DEX pretreatment, with 1521 upregulated (red) and 736 downregulated (blue) mRNAs. B, Heatmap of upregulated (red) and downregulated (green) mRNAs with a clustering tree shown on the left, samples shown in columns and mRNAs shown in rows. C, KEGG analysis of downregulated mRNAs. D–J, Validation of differentially expressed mRNAs associated with the JAK/STAT signaling pathway by qPCR (n = 5). K–N, pSTAT3-Tyr705, pSTAT3-Ser727, and total STAT3 in cardiac tissues (n = 7). Data are presented as the mean ± SD. α2-AR indicates α2-adrenoceptor; DEX, dexmedetomidine; JAK, Janus kinase; KEGG, Kyoto Encyclopedia of Genes and Genomes; mRNA, messenger RNA; qPCR, quantitative polymerase chain reaction; STAT, signal transducer and activator of transcription; TUNEL, TdT-mediated dUTP nick-end labeling.

Article Snippet: After 5% BSA (B24726, ABCONE) blocking, membranes were incubated with primary antibodies, including anti-α2a-AR (1:1000, PA1-30159, Invitrogen), anti-α2b-AR (1:500, sc-390430, Santa Cruz), anti-α2c-AR (1:500, PA1-4518, Invitrogen), anti-STAT3 (1:1000, 12640S), anti-p-STAT3 Y705 (1:2000, 9145S), anti-p-STAT3 S727 (1:1000, 9134S), and anticleaved-caspase 3 (1:1000, 9664S) antibodies from CST, antiGAPDH (1:20,000, 60004-1-Ig), and anti-α-Tubulin antibody (1:20,000, 66031-1-Ig) from Proteintech at 4 °C overnight.

Techniques: Phospho-proteomics, Biomarker Discovery, Real-time Polymerase Chain Reaction, TUNEL Assay, End Labeling

Journal: Anesthesia and Analgesia

Article Title: Dexmedetomidine Pretreatment Protects Against Myocardial Ischemia/Reperfusion Injury by Activating STAT3 Signaling

doi: 10.1213/ANE.0000000000006487

Figure Lengend Snippet: STAT3 phosphorylation is required for the cardioprotective effect of dexmedetomidine. A, CK-MB levels in serum (n = 6). B–D, II-1β , Il-6 , and Tnf-α expression in cardiac tissue (n = 6). E, Immunohistochemistry was performed to reveal the levels of 4-HNE and cleaved caspase 3 in cardiac tissue (n = 3). F and G, TUNEL staining and analysis (n = 3). H–K, pSTAT3-Tyr705, pSTAT3-Ser727, and total STAT3 in cardiac tissues (n = 7). Data are presented as the mean ± SD. Scale bar = 100 μm. 4-HNE indicates 4-hydroxynonenal; CK-MB, creatine kinase-MB isoenzyme; SD, standard deviation; STAT, signal transducer and activator of transcription; TUNEL, TdT-mediated dUTP nick-end labeling.

Article Snippet: After 5% BSA (B24726, ABCONE) blocking, membranes were incubated with primary antibodies, including anti-α2a-AR (1:1000, PA1-30159, Invitrogen), anti-α2b-AR (1:500, sc-390430, Santa Cruz), anti-α2c-AR (1:500, PA1-4518, Invitrogen), anti-STAT3 (1:1000, 12640S), anti-p-STAT3 Y705 (1:2000, 9145S), anti-p-STAT3 S727 (1:1000, 9134S), and anticleaved-caspase 3 (1:1000, 9664S) antibodies from CST, antiGAPDH (1:20,000, 60004-1-Ig), and anti-α-Tubulin antibody (1:20,000, 66031-1-Ig) from Proteintech at 4 °C overnight.

Techniques: Phospho-proteomics, Expressing, Immunohistochemistry, TUNEL Assay, Staining, Standard Deviation, End Labeling