Journal: World Journal of Diabetes

Article Title: Electroacupuncture protects gastric Cajal cells by reducing macrophage pyroptosis in diabetic gastroparesis

doi: 10.4239/wjd.v17.i2.114252

Figure Lengend Snippet: Proposed mechanistic hypothesis in diabetic gastroparesis. In the diabetic gastroparesis setting, M2 macrophages usually recognize and phagocytose apoptotic interstitial cells of Cajal (ICCs). However, upregulation of Gasdermin D triggers pyroptosis, impairing mitochondrial function and reducing phagocytic efficiency. The resulting accumulation of damaged DNA from incompletely cleared ICCs activates the cyclic GMP-AMP synthase-stimulator of interferon genes inflammatory pathway, leading to the release of proinflammatory mediators and further apoptosis of ICCs. The dashed section represents assumptions based on the literature. ICC: Interstitial cells of Cajal; IL: Interleukin; GSDMD-N: Gasdermin D-N terminal domain; NLRP3: NOD-like receptor family pyrin domain containing 3; mtDNA: Mitochondrial DNA; cGAS: Cyclic GMP-AMP synthase; STING: Stimulator of interferon genes; IRF3: Interferon regulatory factor 3; NF-κB: Nuclear factor-κB; TBK1: TANK-binding kinase 1; GSDMD: Gasdermin D.

Article Snippet: IRF3 , 1:100 , Rabbit , Boster, A00165-7.

Techniques: Binding Assay

Journal: World Journal of Diabetes

Article Title: Electroacupuncture protects gastric Cajal cells by reducing macrophage pyroptosis in diabetic gastroparesis

doi: 10.4239/wjd.v17.i2.114252

Figure Lengend Snippet: Effects of electroacupuncture on the cyclic GMP-AMP synthase-stimulator of interferon genes signaling pathway. A: Representative western blot images; B-F: Quantitative analysis of cyclic GMP-AMP synthase, stimulator of interferon genes, p- stimulator of interferon genes, interferon regulatory factor 3, and p-interferon regulatory factor 3 protein expression across groups. Data are presented as mean ± SD ( n = 6). a P < 0.05 vs control group, and b P < 0.05 vs diabetic gastroparesis group. DGP: Diabetic gastroparesis; SEA: Sham-electroacupuncture; LEA: Low-frequency electroacupuncture; HEA: High-frequency electroacupuncture; cGAS: Cyclic GMP-AMP synthase; STING: Stimulator of interferon genes; IRF3: Interferon regulatory factor 3; GAPDH: Glyceraldehyde 3-phosphate dehydrogenase; DM: Diabetes mellitus.

Article Snippet: IRF3 , 1:100 , Rabbit , Boster, A00165-7.

Techniques: Western Blot, Expressing, Control

Journal: Frontiers in Pharmacology

Article Title: Amanita muscaria extract potentiates production of proinflammatory cytokines by dsRNA-activated human microglia

doi: 10.3389/fphar.2023.1102465

Figure Lengend Snippet: AME-1 upregulates TLR3 expression. (A) HMC3 cells were treated with AME-1 (50–5,000 μg/mL) for 3 h, RNA was extracted and analyzed via RT-qPCR. Results are presented as fold-gene expression relative to untreated cells and statistics were calculated using One-Way ANOVA with Dunnett’s multiple comparison post hoc analysis relative to fold-change in GAPDH expression: p ≤ 0.01 (**). Data presented as mean ± SEM for n = 3. HMC3 were treated with AME-1 (50–5,000 µg/mL), LPS (1 µg/m) or poly(I:C) (0.5 µg/mL) for 24 h, the cells were lysed and analyzed via Western blot. (B) TLR3 and IRF3 protein expression and (E, F) corresponding densitometry analysis. (C, D) MDA5 and RIG-I protein expression and (G, H) corresponding densitometry analysis. Densitometry analysis was performed on three blots on the indicated bands and statistical significance was calculated using one-way ANOVA with Tukey post hoc analysis.

Article Snippet: The membrane was subsequently blocked for 1 h at room temperature with 5% skim milk in TBS-T followed by incubation for1hr with 0.4 µg/mL of mouse anti-human TLR3 (sc-32232, Santa Cruz Biotechnology, Dallas, TX, United States), 0.5 µg/mL of mouse anti-human IRF3 (550428, BD biosciences), 2.5 µg/mL of rabbit anti-human MDA5 (33H12L34, Invitrogen), 1 µg/mL rabbit anti-human RIG-1 (PA510297, Invitrogen), or 1 µg/mL of mouse anti-β-actin loading control (PA1-183, Invitrogen) in 3% BSA-TBS-T, and finally incubated for 1 h with goat anti-mouse IRDye 680 (Li-Cor, Lincoln, NE, United States) or goat anti-rabbit IRDye 800 (Li-Cor) diluted 1:15,000 in 3% BSA-TBS-T.

Techniques: Expressing, Quantitative RT-PCR, Western Blot