Journal: Journal of Inflammation Research

Article Title: Discovery and Development of a Novel mPGES-1/5-LOX Dual Inhibitor LFA-9 for Prevention and Treatment of Chronic Inflammatory Diseases

doi: 10.2147/jir.s286110

Figure Lengend Snippet: Figure 5 Anti-inflammatory activity of LFA-9. Paw thickness (A) and percent inhibition (B) using inflammogen (carrageenan (CAR))-induced rat paw edema. (C) Effect of LFA-9 on PGE2, LTB4, PGI2, and TXB2 levels in exudates of carrageenan (CAR))-induced rat paw edema. (D) Effect of LFA-9 on expression of CD68 and TNF-α in CAR- injected paw tissue using immunoblot analysis. (E and F) Effect of LFA-9 on CD68-positive inflammatory cell infiltration and TNF-α expression in CAR-injected paw tissue sections using immunofluorescence technique. P-values ≤0.05 for data sets were considered significant. (NS) Not significant (P>0.05); *(P ≤ 0.05); **(P ≤ 0.01); *** (P ≤ 0.001); **** (P ≤ 0.0001). (G) Schematic representation of mechanism of action of LFA-9 for its anti-inflammatory activity with safety.

Article Snippet: After incubation with primary antibody [Alexa Fluor 700-conjugated CD68 (1:50 dilution) (Novus Biologicals (NB600985)) and TNF-α (1:100 dilution) (Santa Cruz)] overnight at 4°C, slides were labeled with Alexa Fluor 488 dye-conjugated secondary antibody (for TNF-α) and mounted with ProLong Gold (Thermo Fisher Scientific).

Techniques: Activity Assay, Inhibition, Expressing, Injection, Western Blot, Immunofluorescence

Journal: Communications Biology

Article Title: MicroRNA-221-3p inhibits the inflammatory response of keratinocytes by regulating the DYRK1A/STAT3 signaling pathway to promote wound healing in diabetes

doi: 10.1038/s42003-024-05986-0

Figure Lengend Snippet: Representative immunohistochemistry images ( a , b ) and summary data ( c , b ) showing myeloperoxidase (MPO) ( a , c ) and CD68 ( b , d ) expression levels on days 1, 3, 7, and 11 after wound formation in diabetic mice injected with miRNA mimic negative control (DM-Ctrl) or miR-221-3p agomir (DM-miR-221-3p). Density of positive cells was calculated using the following formula: number of positive cells/area size ( n = 4–6). Scale bars, 50 μm. Summary data showing relative mRNA expression levels of IL-1β, IL-6, IL-8 , and TNF-α at the edge of the skin wound on days 1 ( e ), 7 ( f ) and 11 ( g ) ( n = 6–10). h Summary data showing relative mRNA expression levels of CD86 and CD206 at the edge of the skin wound on days 11 ( n = 6–10). The mRNA levels were normalized to 18S ribosomal RNA. Data in c-h are presented as mean ± SEM. Solid circles represent DM-Ctrl group and squares, DM-miR-221-3p group. * P < 0.05, ** P < 0.01 for DM-miR-221-3p vs. DM-Ctrl.

Article Snippet: Sections were then incubated overnight at 4 °C with primary antibody CD68 (NB600-985, Novus Biologicals, USA), MPO (AF3667, Abcam, UK) or DYRK1A (DF3270, Affinity Biosciences, China) followed by incubation with a secondary antibody conjugated with streptavidin-horseradish peroxidase.

Techniques: Immunohistochemistry, Expressing, Injection, Negative Control

Journal: Communications Biology

Article Title: MicroRNA-221-3p inhibits the inflammatory response of keratinocytes by regulating the DYRK1A/STAT3 signaling pathway to promote wound healing in diabetes

doi: 10.1038/s42003-024-05986-0

Figure Lengend Snippet: Representative immunohistochemistry images and summary data showing myeloperoxidase (MPO) ( a ), CD68 ( b ) and DYRK1A ( c ) expression levels in skin wound tissues of wild type (WT) and Mir221 knockout (KO) mice. Data are presented as mean ± SEM ( n = 5–8). Scale bars, 50 μm in ( a , b ), 5 μm in ( c ). d Representative images and summary data showing expression levels of phosphorylated (p)-STAT3 (Tyr705), p-STAT3 (Ser727), total STAT3, and DYRK1A in skin wound tissues of WT and Mir221 KO mice. Vertically stacked strips of bands in a figure are not derived from the same membrane in any case. Data are presented as mean ± SEM ( n = 5), * P < 0.05.

Article Snippet: Sections were then incubated overnight at 4 °C with primary antibody CD68 (NB600-985, Novus Biologicals, USA), MPO (AF3667, Abcam, UK) or DYRK1A (DF3270, Affinity Biosciences, China) followed by incubation with a secondary antibody conjugated with streptavidin-horseradish peroxidase.

Techniques: Immunohistochemistry, Expressing, Knock-Out, Derivative Assay, Membrane

Journal: Stem cell research & therapy

Article Title: Comparison of reversal of rat pulmonary fibrosis of nintedanib, pirfenidone, and human umbilical mesenchymal stem cells from Wharton's jelly.

doi: 10.1186/s13287-020-02012-y

Figure Lengend Snippet: Fig. 7 Comparison of the activation of macrophage and the expression of TLR-4 of the left lung in rats with PF. Left lung sections were obtained from the Normal, BLM, BLM + Nintedanib, BLM + Pirfenidone, and BLM + HUMSCs groups on day 49 and were subjected to immunohistochemical stain with anti-ED1 antibody for macrophage labeling. From low to high magnifications, the number of macrophages with a relatively small morphology increased in the BLM group. Transplantation of HUMSCs triggered the activation of macrophage with a relatively bigger size. The patterns of macrophages were similar between the rest treatment groups and the BLM group (a, b). Next, anti-CD86 antibody was applied to label M1 macrophage, which revealed that the number of M1 macrophages increased in the left lungs of the BLM, BLM + Nintedanib, and BLM + Pirfenidone groups (c, e). Additionally, anti-CD163 antibody was used to mark M2 macrophage, which showed that the number of M2 macrophages elevated in the left lung of the BLM, BLM + Nintedanib, and BLM + Pirfenidone groups; the number of M2 macrophages significantly increased in the BLM + HUMSCs group (d, f). Moreover, the expression level of MMP-9 (g) and TLR-4 (h) in the left lung was quantified with Western blotting, results showed that transplantation of HUMSCs resulted in alternative macrophage activation and increased MMP-9 and TLR-4 expressions in the left lung of rats with PF. n = 3 in each group. ✽: vs the Normal group, p < 0.05. ♦: vs the BLM + HUMSCs group, p < 0.05

Article Snippet: Lung slices (with recovered antigens) were reacted with primary antibodies (mouse anti-α-smooth muscle actin antibody for myofibroblast [SMA, Sigma A2547]; mouse anti-ED1 antibody for total macrophage [Millipore MAB1435]; rabbit anti-CD86 antibody for M1 macrophage [Proteintech, 13395-1-AP]; mouse anti-CD163 antibody for M2 macrophage [BioRad MCA342R]; rabbit anti-NADPH oxidase antibody [Abcam, ab133303]; mouse anti-catalase antibody [Abnova, H00000847A01]) at 4 °C for 12–18 h and then reacted with secondary antibodies.

Techniques: Comparison, Activation Assay, Expressing, Immunohistochemical staining, Staining, Labeling, Transplantation Assay, Western Blot