Journal: Neural Plasticity

Article Title: Mahuang-Fuzi-Xixin Decoction Reverses Depression-Like Behavior in LPS-Induced Mice by Regulating NLRP3 Inflammasome and Neurogenesis

doi: 10.1155/2019/1571392

Figure Lengend Snippet: MFX-M alleviates the NLPR3 inflammasome activation in the hippocampus of LPS-stimulated mice. Representative blots and statistical graphs of relative protein expression of NLRP3 (a), ASC (b), cl-caspase-1, pro-caspase-1 (c), IL-1 β (d), and TXNIP (e). (f) Frozen sections of brain tissue were subjected to immunofluorescence staining. The hippocampus sections were incubated with anti-GFAP antibody (green), anti-IL-1 β (red), and DAPI (blue) and were observed by a laser scanning confocal microscope. Scale bar = 40 μ m. Data were the mean ± S.E.M. ( n = 3 experimental replicates/group). ∗ p < 0.05 and ∗∗ p < 0.01 compared to the control group. # p < 0.05 and ## p < 0.01 compared to the LPS group.

Article Snippet: Rabbit polyclonal anti-NLRP3 (ab214185), rabbit polyclonal anti-IL-1 β (ab9722), rabbit monoclonal anti-TXNIP (ab188865), rabbit monoclonal anti-DCX (ab207175), rabbit polyclonal anti-DCX (ab18723), and rabbit monoclonal anti-BDNF (ab108319) were acquired from Abcam.

Techniques: Activation Assay, Expressing, Immunofluorescence, Staining, Incubation, Microscopy

Journal: Annals of Translational Medicine

Article Title: Anti-inflammatory effects of artesunate on atherosclerosis via miR-16-5p and TXNIP regulation of the NLRP3 inflammasome

doi: 10.21037/atm-21-4939

Figure Lengend Snippet: Reduced expression of miR-16-5p and elevated expression of TXNIP in PB are indicative of severity of coronary AS. CD14+ monocytes were isolated from PB of 115 CHD patients and 33 non-CHD patients confirmed by coronary angiography. (A) The protein level of IL-1β and IL-18 in CD14+ monocytes assessed by ELISA. (B) Pearson’s correlation analysis between Gensini scores and protein level of IL-1β and IL-18 in CD14+ monocytes. (C) The expression of miR-16-5p and TXNIP in CD14+ monocytes determined by RT-qPCR. (D) Pearson’s correlation analysis between the Gensini scores and the expression of miR-16-5p and TXNIP in CD14+ monocytes. * (relative to non-CHD patients), indicates P<0.05 by unpaired t-test. PB, peripheral blood; CHD, coronary heart disease; IL-1β, interleukin-1β; ELISA, enzyme-linked immunosorbent assay; RT-qPCR, reverse transcription quantitative polymerase chain reaction.

Article Snippet: Primary antibodies were as follows, anti-rabbit IL-1β polyclonal antibody (1:1,000, ab2105, Abcam, Cambridge, UK), anti-mouse mature IL-1β monoclonal antibody (1:500, Asp116, Cell Signaling Technology, Beverly, MA, USA), anti- rabbit-TXNIP monoclonal antibody (1:1,000, ab188865, Abcam), anti-rabbit NLRP3 polyclonal antibody (1:500, ab214185, Abcam), anti-rabbit recombinant caspase-1 monoclonal antibody (1:1,000, ab207802, Abcam), anti-rabbit IL-18 polyclonal antibody (1:1,000, ab191152, Abcam), anti-mouse mature IL-18 monoclonal antibody (1:1,000, D044-3M2, MBL international corporation, Woburn, UK), and anti-mouse β-actin monoclonal antibody (1:5,000, ab8225, Abcam).

Techniques: Expressing, Isolation, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Real-time Polymerase Chain Reaction

Journal: Annals of Translational Medicine

Article Title: Anti-inflammatory effects of artesunate on atherosclerosis via miR-16-5p and TXNIP regulation of the NLRP3 inflammasome

doi: 10.21037/atm-21-4939

Figure Lengend Snippet: Artesunate modulates TXNIP-mediated activation of NLRP3 inflammasome. (A) The protein expression of TXNIP in macrophages determined by immunoblotting. (B) The interaction between TXNIP and NLRP3 in macrophages determined by CO-IP. (C) The expression of IL-1β and IL-18 in macrophage culture supernatant assessed by ELISA. * (relative to control) and # (relative to ATP), indicate P<0.05 by one-way ANOVA/Tukey’s test. CO-IP, coimmunoprecipitation; IL-1β, interleukin-1β; enzyme-linked immunosorbent assay; ANOVA, analysis of variance.

Article Snippet: Primary antibodies were as follows, anti-rabbit IL-1β polyclonal antibody (1:1,000, ab2105, Abcam, Cambridge, UK), anti-mouse mature IL-1β monoclonal antibody (1:500, Asp116, Cell Signaling Technology, Beverly, MA, USA), anti- rabbit-TXNIP monoclonal antibody (1:1,000, ab188865, Abcam), anti-rabbit NLRP3 polyclonal antibody (1:500, ab214185, Abcam), anti-rabbit recombinant caspase-1 monoclonal antibody (1:1,000, ab207802, Abcam), anti-rabbit IL-18 polyclonal antibody (1:1,000, ab191152, Abcam), anti-mouse mature IL-18 monoclonal antibody (1:1,000, D044-3M2, MBL international corporation, Woburn, UK), and anti-mouse β-actin monoclonal antibody (1:5,000, ab8225, Abcam).

Techniques: Activation Assay, Expressing, Western Blot, Co-Immunoprecipitation Assay, Enzyme-linked Immunosorbent Assay

Journal: Annals of Translational Medicine

Article Title: Anti-inflammatory effects of artesunate on atherosclerosis via miR-16-5p and TXNIP regulation of the NLRP3 inflammasome

doi: 10.21037/atm-21-4939

Figure Lengend Snippet: Artesunate blocks the NLRP3 inflammasome via inhibition of miR-16-5p-mediated TXNIP. (A,E,F) The expression of miR-16-5p in macrophages determined by RT-qPCR. (B,G) The protein expression of TXNIP in macrophages, pro-caspase-1, -IL-1β and -IL-18 and their mature forms released in macrophages assessed by immunoblotting. (C,I) The expression of IL-1β and IL-18 in macrophages culture supernatants assessed by ELISA. (D,H) ASC production in macrophages determined by ASC. * (relative to mimic-NC + PBS) and # (relative to mimic-NC + ATP) indicates P<0.05 in panel (A-D); * (relative to control), # (relative to 2.5 µmol/L artesunate), and & (relative to 5 µmol/L artesunate) indicates P<0.05 in panel (E); * (relative to inhibitor-NC + PBS), # (relative to inhibitor-NC + PBS), and & (relative to inhibitor-NC + artesunate + ATP) indicates P<0.05 in panel (F-I). Statistical significance was confirmed by one-way ANOVA/Tukey’s test. RT-qPCR, reverse transcription quantitative polymerase chain reaction; IL-1β, interleukin-1β; enzyme-linked immunosorbent assay; ANOVA, analysis of variance; ATP, adenosine triphosphate; ASC, apoptosis-associated speck-like protein containing caspase recruitment domain.

Article Snippet: Primary antibodies were as follows, anti-rabbit IL-1β polyclonal antibody (1:1,000, ab2105, Abcam, Cambridge, UK), anti-mouse mature IL-1β monoclonal antibody (1:500, Asp116, Cell Signaling Technology, Beverly, MA, USA), anti- rabbit-TXNIP monoclonal antibody (1:1,000, ab188865, Abcam), anti-rabbit NLRP3 polyclonal antibody (1:500, ab214185, Abcam), anti-rabbit recombinant caspase-1 monoclonal antibody (1:1,000, ab207802, Abcam), anti-rabbit IL-18 polyclonal antibody (1:1,000, ab191152, Abcam), anti-mouse mature IL-18 monoclonal antibody (1:1,000, D044-3M2, MBL international corporation, Woburn, UK), and anti-mouse β-actin monoclonal antibody (1:5,000, ab8225, Abcam).

Techniques: Inhibition, Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction

Journal: Annals of Translational Medicine

Article Title: Anti-inflammatory effects of artesunate on atherosclerosis via miR-16-5p and TXNIP regulation of the NLRP3 inflammasome

doi: 10.21037/atm-21-4939

Figure Lengend Snippet: Artesunate inhibits atherosclerotic plaque formation by its anti-inflammatory effects in vivo. (A) HE staining on the plaque formation in arterial tissues of rats. (B) The protein expression of IL-1β and IL-18 in arterial tissues of rats determined by ELISA. (C) The expression of miR-16-5p and TXNIP in arterial tissues of rats assessed by RT-qPCR. (D) The level of blood liquid in in arterial tissues of rats determined by ELISA. * (relative to control, # (relative to AS rats), & (relative to low-dose artesunate), ^ (relative to AS + medium-dose artesunate) and △ (relative to AS + high-dose artesunate + antagomir-16-5p-NC), indicates P<0.05 by one-way ANOVA/Tukey’s post. Each rat group consists of 10 rats. HE, hematoxylin-eosin; IL-1β, interleukin-1β; enzyme-linked immunosorbent assay; ANOVA, analysis of variance; RT-qPCR, reverse transcription quantitative polymerase chain reaction.

Article Snippet: Primary antibodies were as follows, anti-rabbit IL-1β polyclonal antibody (1:1,000, ab2105, Abcam, Cambridge, UK), anti-mouse mature IL-1β monoclonal antibody (1:500, Asp116, Cell Signaling Technology, Beverly, MA, USA), anti- rabbit-TXNIP monoclonal antibody (1:1,000, ab188865, Abcam), anti-rabbit NLRP3 polyclonal antibody (1:500, ab214185, Abcam), anti-rabbit recombinant caspase-1 monoclonal antibody (1:1,000, ab207802, Abcam), anti-rabbit IL-18 polyclonal antibody (1:1,000, ab191152, Abcam), anti-mouse mature IL-18 monoclonal antibody (1:1,000, D044-3M2, MBL international corporation, Woburn, UK), and anti-mouse β-actin monoclonal antibody (1:5,000, ab8225, Abcam).

Techniques: In Vivo, Staining, Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Real-time Polymerase Chain Reaction

Journal: Annals of Translational Medicine

Article Title: Anti-inflammatory effects of artesunate on atherosclerosis via miR-16-5p and TXNIP regulation of the NLRP3 inflammasome

doi: 10.21037/atm-21-4939

Figure Lengend Snippet: The mechanism graph of the regulatory network and function of artesunate on AS. Artesunate blocks TXNIP-mediated NLRP3 inflammasome via the upregulation of miR-16-5p to exert anti-inflammatory effects on AS.

Article Snippet: Primary antibodies were as follows, anti-rabbit IL-1β polyclonal antibody (1:1,000, ab2105, Abcam, Cambridge, UK), anti-mouse mature IL-1β monoclonal antibody (1:500, Asp116, Cell Signaling Technology, Beverly, MA, USA), anti- rabbit-TXNIP monoclonal antibody (1:1,000, ab188865, Abcam), anti-rabbit NLRP3 polyclonal antibody (1:500, ab214185, Abcam), anti-rabbit recombinant caspase-1 monoclonal antibody (1:1,000, ab207802, Abcam), anti-rabbit IL-18 polyclonal antibody (1:1,000, ab191152, Abcam), anti-mouse mature IL-18 monoclonal antibody (1:1,000, D044-3M2, MBL international corporation, Woburn, UK), and anti-mouse β-actin monoclonal antibody (1:5,000, ab8225, Abcam).

Techniques: