Journal: Scientific Reports

Article Title: Remimazolam alleviates cerebral ischemia–reperfusion injury of rats by inhibiting NF-κB/NLRP3 inflammasome pyroptosis

doi: 10.1038/s41598-025-31205-9

Figure Lengend Snippet: Remimazolam reduces cortical neuronal NLRP3/ASC/Caspase-1 inflammasome-dependent pyroptosis by inhibiting NF-κB pathway activation. ( a ) The protein expressions of the NF-κB p65, NLRP3, and ASC were significantly reduced in the JSH-23 group, however, the protein expression of the NF-κB p65 was not significantly changed in the MCC950 group compared with the MCAO group. ( b ) Fluorescence intensity (green staining) of NF-κB p65, NLRP3, GSDMD, and Caspase-1 p20. Nuclei were stained with DAPI. Bar: 20 μm. ( c , d ) the proteins expressions of NF-κB p65, NLRP3, ASC, and caspase-1 p20 and the mRNAs of NF-κB p65, NLRP3, ASC, GSDMD, and caspase-1 p20 in vitro. ( e ) IL-1β secretion by ELISA assays both in vitro and in vivo studies. All tests were repeated independently three times. Data are presented as Mean ± SEM. ns: no significant; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001: significant as compared to Sham group or control group; # p < 0.05, ## p < 0.01, ### p < 0.001, and #### p < 0.0001: significant as compared to MCAO group or OGD group.

Article Snippet: MCC950 (Selleck, S7809, USA), a select NLRP3 inhibition, was administered as the group MCC950 , .

Techniques: Activation Assay, Expressing, Fluorescence, Staining, In Vitro, Enzyme-linked Immunosorbent Assay, In Vivo, Control

Journal: BMC Complementary Medicine and Therapies

Article Title: Inhibitory effects of the flavonoids extracted from Pollen Typhae on palmitic acid-induced NLRP3 inflammasome activation in macrophages involving AMPK-mediated lipid metabolism

doi: 10.1186/s12906-025-05024-4

Figure Lengend Snippet: Pollen Typhae flavonoids attenuate PA-induced NLRP3 inflammasome activation in macrophages. ( A ) After pretreatment with or without LPS (1 ng/ml) for 3 h, RAW264.7 macrophages were stimulated with PA (0.125 ~ 0.5 mM) for 16 h, IL-1β and IL-18 in culture supernatants were analyzed by ELISA. ( B ) The macrophages were pretreated with LPS (1 ng/ml) for 3 h, followed by PA (0.5 mM) treatment for 16 h, the protein expression of NLRP3, Caspase-1, and Caspase-1 p10 was analyzed by western blotting. The uncropped blots were presented in Supplementary Fig. . ( C ) RAW264.7 macrophages were treated with PTF (0 ~ 0.5 mg/ml), TYP (0 ~ 100 µM) or I3ON (0 ~ 100 µM) for 16 and 20 h, the cell viability was detected by CCK-8 assay. * P < 0.05, ** P < 0.01 versus 0.0 mg/ml PTF (0.0 µM TYP or I3ON). ( D ) The macrophages were pretreated with LPS (1 ng/ml) or plus different concentrations of PTF (0.03125 ~ 0.5 mg/ml), TYP (10 ~ 100 µM), I3ON (10 ~ 100 µM) for 3 h before exposure to PA (0.5 mM) for 16 h, IL-1β and IL-18 in culture supernatants were checked by ELISA. ( E and F ) The macrophages were pretreated with LPS (1 ng/ml) or plus PTF (0.5 mg/ml), TYP (100 µM), AICAR (2 mM), MCC950 (10 µM) for 3 h followed by exposure to PA (0.5 mM) for 16 h, the protein (E) and gene expression (F) of NLRP3, Caspase-1 and IL-1β were checked by western blotting and Real-time PCR, respectively. The uncropped/unedited blots were presented in Supplementary Fig. . Each column shows the mean with SD ( n = 3–7). * P < 0.05, ** P < 0.01 versus LPS (1 ng/ml); # P < 0.05, ## P < 0.01 versus LPS (1 ng/ml) plus PA (0.5 mM)

Article Snippet: The macrophages were pre-exposed to LPS (1 ng/ml) or/and PTF (0.5 mg/ml), TYP (100 μM), AICAR (2 mM, Beyotime, Shanghai, China), MCC950 (10 μM, Cell Signaling Technology, Danvers, MA, USA), PF-05175157 (10 μM, GlpBio Technology, Montclair, CA, USA) for 3 h before treatment with PA (0.5 mM) for 16 h. The cells were harvested to obtain total protein.

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, CCK-8 Assay, Gene Expression, Real-time Polymerase Chain Reaction

Journal: BMC Complementary Medicine and Therapies

Article Title: Inhibitory effects of the flavonoids extracted from Pollen Typhae on palmitic acid-induced NLRP3 inflammasome activation in macrophages involving AMPK-mediated lipid metabolism

doi: 10.1186/s12906-025-05024-4

Figure Lengend Snippet: Effects of PTF, AMPK agonist, ACC inhibitor and antioxidant on PA-induced Caspase-1 activity in macrophages. RAW264.7 macrophages were pretreated with LPS (1 ng/ml) or plus PTF (0.5 mg/ml), TYP (100 µM), PF-05175157 (10 µM), AICAR (2 mM), Compound C (10 µM), NAC (10 mM), MCC950 (10 µM) for 3 h before treatment with PA (0.5 mM) for 16 h, the Caspase-1 activity in culture supernatants ( A ) and the macrophages ( B ) was analyzed by bioluminescent method. Each column shows the mean with SD ( n = 3). * P < 0.05, ** P < 0.01 versus LPS (1 ng/ml); # P < 0.05, ## P < 0.01 versus LPS (1 ng/ml) plus PA (0.5 mM); § P < 0.05 versus LPS (1 ng/ml) plus PA (0.5 mM) and PTF (0.5 mg/ml)

Article Snippet: The macrophages were pre-exposed to LPS (1 ng/ml) or/and PTF (0.5 mg/ml), TYP (100 μM), AICAR (2 mM, Beyotime, Shanghai, China), MCC950 (10 μM, Cell Signaling Technology, Danvers, MA, USA), PF-05175157 (10 μM, GlpBio Technology, Montclair, CA, USA) for 3 h before treatment with PA (0.5 mM) for 16 h. The cells were harvested to obtain total protein.

Techniques: Activity Assay

Journal: Immunity, Inflammation and Disease

Article Title: The role of autophagy in SIM mediated anti‐inflammatory osteoclastogenesis through NLRP3 signaling pathway

doi: 10.1002/iid3.1145

Figure Lengend Snippet: Simvastatin (SIM) suppresses lipopolysaccharide (LPS)‐induced osteoclast (OC)‐associated gene expression. RAW264.7 cells were stimulated with NOD‐like receptor family pyrin Domain‐containing protein 3 (NLRP3) inhibitor MCC950 (30 µM) and SIM, then treated them with 100 ng/mL of LPS for 12 h. (A–C) The expression of OC‐specific genes ( RANK , CTSK , and c‐Fos ) were detected by reverse transcription‑quantitative polymerase chain reaction (RT‐qPCR). Bars represent the mean ± SD of three independent experiments. ## p < .01 and ### p < .001 compared with control group; * p < .05, ** p < .01, and *** p < .001 compared with LPS‐treated group.

Article Snippet: MCC950 (CP‐456773, Cat# GC31644) was purchased from Glpbio Technology.

Techniques: Expressing, Polymerase Chain Reaction, Quantitative RT-PCR, Control

Journal: Immunity, Inflammation and Disease

Article Title: The role of autophagy in SIM mediated anti‐inflammatory osteoclastogenesis through NLRP3 signaling pathway

doi: 10.1002/iid3.1145

Figure Lengend Snippet: MCC950 is involved in the inhibition of Simvastatin (SIM) on autophagy‐related proteins in lipopolysaccharide (LPS)‐treated RAW264.7 macrophages. (A) MCC950 was pretreated into LPS‐stimulated RAW264.7 macrophages in the presence or absence of SIM. (B–D) The band intensities were quantified using ImageJ software. Bars represent the mean ± SD of three independent experiments. # p < .05 and ## p < .01 compared with control group; * p < .05 and ** p < .01 compared with LPS‐treated group.

Article Snippet: MCC950 (CP‐456773, Cat# GC31644) was purchased from Glpbio Technology.

Techniques: Inhibition, Software, Control

Journal: Frontiers in Cellular Neuroscience

Article Title: Electroacupuncture Ameliorates Mechanical Allodynia of a Rat Model of CRPS-I via Suppressing NLRP3 Inflammasome Activation in Spinal Cord Dorsal Horn Neurons

doi: 10.3389/fncel.2022.826777

Figure Lengend Snippet: EA's anti-allodynic effect on CPIP rats is mimicked by pharmacological blocking NLRP3 inflammasome. (A) Experimental protocol. (B) 50% PWT following EA or MCC950 intervention (30 μg/rat/day, i.t.). (C) Normalized AUC deduced from (B) . (D) Immunostaining showing the effect of EA/MCC950 intervention on astrocyte (Upper panels, marked with GFAP) and microglia overactivation in SCDH. (E,F) Summary of the mean fluorescence intensity of GFAP (E) and OX42 (F) , normalized with the value of sham group. n = 5 rats/group. Scale bar indicates 100 μm. ** p < 0.01 # p < 0.05, ## p < 0.01. Two-way ANOVA with Tukey's post-hoc test was applied in panel B. One-way ANOVA with Tukey's post-hoc test was applied in (C,E,F) . NS, no significance.

Article Snippet: NLPR3 specific antagonist MCC950 and the activator nigericin were purchased from APExBIO Technology (USA).

Techniques: Blocking Assay, Immunostaining, Fluorescence