Journal: Molecular Medicine Reports

Article Title: Glabridin attenuates LTA-induced alveolar macrophage migration via activation of the Nrf2/HO-1 pathway

doi: 10.3892/mmr.2025.13758

Figure Lengend Snippet: Pharmacological and genetic inhibition of Nrf2 attenuates GBD-induced HO-1 expression. MH-S cells were pretreated with the indicated compounds for 30 min and then stimulated with LTA. (A) Immunofluorescence analysis of HO-1 expression 6 h poststimulation. Scale bar, 10 µm. (B) Immunofluorescence image evaluation of HO-1 mean fluorescence intensity. (C) HO-1 mRNA expression 6 h poststimulation, determined through RT-qPCR. (D and E) Effect of Nrf2 knockdown on GBD-induced HO-1 expression. MH-S cells were transfected with control siRNA (siNC) or Nrf2 siRNA (siNrf2) for 6 h, followed by the indicated treatments. HO-1 mRNA levels were measured by RT-qPCR. Data are presented as means ± SD. ***P<0.001 vs. DMSO+LTA (B-C) or si-Ctrl+DMSO (D and E); ## P<0.01 and ### P<0.001 vs. GBD+LTA (B-C) or si-Ctrl+DMSO+LTA (E); †† P<0.01 and ††† P<0.001 vs. ML385+LTA (B-C) or si-Ctrl+GBD+LTA (E). Nrf2, nuclear factor erythroid 2-related factor 2; GBD, glabridin; LTA, lipoteichoic acid; HO-1, heme oxygenase-1; RT-qPCR, reverse transcription-quantitative PCR; si, short interfering; DMSO, dimethyl sulfoxide.

Article Snippet: Cells were then transfected with either a small interfering (si)RNA targeting Nrf2 [ Nfe2l2 siRNA: 5′-AGCAUUUUAACAUGUUAACAG-3′ (sense) and 5′-GUUAACAUGUUAAAAUGCUAU-3′ (antisense)] or a negative control siRNA [si-Ctrl: 5′-UUCUCCGAACGUGUCACGUTT-3′ (sense) and reverse 5′-ACGUGACACGUUCGGAGAATT-3′ (antisense); cat. no. HY-RS09246; MedChemExpress], using a transfection reagent (cat. no. HY-K2017, MedChemExpress) and 50 nM siRNA diluted in serum-free medium, according to the manufacturer's instructions.

Techniques: Inhibition, Expressing, Immunofluorescence, Fluorescence, Quantitative RT-PCR, Knockdown, Transfection, Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: Glabridin attenuates LTA-induced alveolar macrophage migration via activation of the Nrf2/HO-1 pathway

doi: 10.3892/mmr.2025.13758

Figure Lengend Snippet: GBD activates Nrf2 signaling in response to LTA. MH-S cells were pretreated with GBD for 30 min and then stimulated with LTA. (A) Confocal images indicating Nrf2 nuclear translocation 3 h poststimulation. Scale bar, 50 µm. (B) Time-course of Nrf2 mRNA expression after LTA stimulation (0, 2, 4, 6, and 8 h). (C) Nrf2 mRNA expression was determined through reverse transcription-quantitative PCR 2 h poststimulation, as described in the materials and methods section. (D) Western blot analysis of Nrf2 protein levels 3 h after LTA stimulation. Data are presented as means±SD. *P<0.05, **P<0.01 and ***P<0.001 vs. 0 h or DMSO; ## P<0.01 and ### P<0.001 vs. DMSO+LTA. GBD glabridin; Nrf2, nuclear factor erythroid 2-related factor 2; LTA, lipoteichoic acid; DMSO, dimethyl sulfoxide.

Article Snippet: Cells were then transfected with either a small interfering (si)RNA targeting Nrf2 [ Nfe2l2 siRNA: 5′-AGCAUUUUAACAUGUUAACAG-3′ (sense) and 5′-GUUAACAUGUUAAAAUGCUAU-3′ (antisense)] or a negative control siRNA [si-Ctrl: 5′-UUCUCCGAACGUGUCACGUTT-3′ (sense) and reverse 5′-ACGUGACACGUUCGGAGAATT-3′ (antisense); cat. no. HY-RS09246; MedChemExpress], using a transfection reagent (cat. no. HY-K2017, MedChemExpress) and 50 nM siRNA diluted in serum-free medium, according to the manufacturer's instructions.

Techniques: Translocation Assay, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot

Journal: Molecular Medicine Reports

Article Title: Glabridin attenuates LTA-induced alveolar macrophage migration via activation of the Nrf2/HO-1 pathway

doi: 10.3892/mmr.2025.13758

Figure Lengend Snippet: GBD increases HO-1 expression through Nrf2 signaling under LTA challenge. MH-S cells were pretreated with GBD for 30 min and then stimulated with LTA. (A) Confocal images of HO-1 expression 6 h poststimulation. Scale bar, 50 µm. (B) Time-course of HO-1 mRNA expression after LTA stimulation (0, 2, 4, 6, and 8 h). (C) HO-1 mRNA expression was determined by reverse transcription-quantitative PCR. (D) HO-1 protein expression analyzed through western blotting 6 h poststimulation. Data are presented as means±SD (n=4). *P<0.05 and ***P<0.001 vs. 0 h or DMSO; ### P<0.001 vs. DMSO+LTA. GBD glabridin; HO-1, heme oxygenase-1; Nrf2, nuclear factor erythroid 2-related factor 2; LTA, lipoteichoic acid; DMSO, dimethyl sulfoxide.

Article Snippet: Cells were then transfected with either a small interfering (si)RNA targeting Nrf2 [ Nfe2l2 siRNA: 5′-AGCAUUUUAACAUGUUAACAG-3′ (sense) and 5′-GUUAACAUGUUAAAAUGCUAU-3′ (antisense)] or a negative control siRNA [si-Ctrl: 5′-UUCUCCGAACGUGUCACGUTT-3′ (sense) and reverse 5′-ACGUGACACGUUCGGAGAATT-3′ (antisense); cat. no. HY-RS09246; MedChemExpress], using a transfection reagent (cat. no. HY-K2017, MedChemExpress) and 50 nM siRNA diluted in serum-free medium, according to the manufacturer's instructions.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot

Journal: Molecular Medicine Reports

Article Title: Glabridin attenuates LTA-induced alveolar macrophage migration via activation of the Nrf2/HO-1 pathway

doi: 10.3892/mmr.2025.13758

Figure Lengend Snippet: ML385 inhibits GBD-induced Nrf2 activation in LTA-stimulated MH-S cells. MH-S cells were pretreated with DMSO (0.1%), GBD (20 µM), or ML385 (5 µM) for 30 min and then stimulated with LTA. (A) Confocal microscopy of Nrf2 nuclear localization 3 h poststimulation. Scale bar, 10 µm. (B) Quantification of Nrf2 nuclear accumulation based on nuclear mean fluorescence intensity. (C) Nrf2 mRNA expression 2 h poststimulation, analyzed through reverse transcription-quantitative PCR. Data are presented as means ± SD. ***P<0.001 vs. DMSO+LTA; ## P<0.01 vs. GBD+LTA; †† P<0.01 and ††† P<0.001 vs. ML385+LTA. GBD glabridin; Nrf2, nuclear factor erythroid 2-related factor 2; LTA, lipoteichoic acid; DMSO, dimethyl sulfoxide.

Article Snippet: Cells were then transfected with either a small interfering (si)RNA targeting Nrf2 [ Nfe2l2 siRNA: 5′-AGCAUUUUAACAUGUUAACAG-3′ (sense) and 5′-GUUAACAUGUUAAAAUGCUAU-3′ (antisense)] or a negative control siRNA [si-Ctrl: 5′-UUCUCCGAACGUGUCACGUTT-3′ (sense) and reverse 5′-ACGUGACACGUUCGGAGAATT-3′ (antisense); cat. no. HY-RS09246; MedChemExpress], using a transfection reagent (cat. no. HY-K2017, MedChemExpress) and 50 nM siRNA diluted in serum-free medium, according to the manufacturer's instructions.

Techniques: Activation Assay, Confocal Microscopy, Fluorescence, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: Glabridin attenuates LTA-induced alveolar macrophage migration via activation of the Nrf2/HO-1 pathway

doi: 10.3892/mmr.2025.13758

Figure Lengend Snippet: Pharmacological and genetic inhibition of Nrf2 attenuates GBD-induced HO-1 expression. MH-S cells were pretreated with the indicated compounds for 30 min and then stimulated with LTA. (A) Immunofluorescence analysis of HO-1 expression 6 h poststimulation. Scale bar, 10 µm. (B) Immunofluorescence image evaluation of HO-1 mean fluorescence intensity. (C) HO-1 mRNA expression 6 h poststimulation, determined through RT-qPCR. (D and E) Effect of Nrf2 knockdown on GBD-induced HO-1 expression. MH-S cells were transfected with control siRNA (siNC) or Nrf2 siRNA (siNrf2) for 6 h, followed by the indicated treatments. HO-1 mRNA levels were measured by RT-qPCR. Data are presented as means ± SD. ***P<0.001 vs. DMSO+LTA (B-C) or si-Ctrl+DMSO (D and E); ## P<0.01 and ### P<0.001 vs. GBD+LTA (B-C) or si-Ctrl+DMSO+LTA (E); †† P<0.01 and ††† P<0.001 vs. ML385+LTA (B-C) or si-Ctrl+GBD+LTA (E). Nrf2, nuclear factor erythroid 2-related factor 2; GBD, glabridin; LTA, lipoteichoic acid; HO-1, heme oxygenase-1; RT-qPCR, reverse transcription-quantitative PCR; si, short interfering; DMSO, dimethyl sulfoxide.

Article Snippet: Cells were then transfected with either a small interfering (si)RNA targeting Nrf2 [ Nfe2l2 siRNA: 5′-AGCAUUUUAACAUGUUAACAG-3′ (sense) and 5′-GUUAACAUGUUAAAAUGCUAU-3′ (antisense)] or a negative control siRNA [si-Ctrl: 5′-UUCUCCGAACGUGUCACGUTT-3′ (sense) and reverse 5′-ACGUGACACGUUCGGAGAATT-3′ (antisense); cat. no. HY-RS09246; MedChemExpress], using a transfection reagent (cat. no. HY-K2017, MedChemExpress) and 50 nM siRNA diluted in serum-free medium, according to the manufacturer's instructions.

Techniques: Inhibition, Expressing, Immunofluorescence, Fluorescence, Quantitative RT-PCR, Knockdown, Transfection, Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: Glabridin attenuates LTA-induced alveolar macrophage migration via activation of the Nrf2/HO-1 pathway

doi: 10.3892/mmr.2025.13758

Figure Lengend Snippet: GBD inhibits LTA-induced macrophage migration through Nrf2 activation. MH-S cells were pretreated with the indicated compounds for 30 min and then stimulated with LTA. (A) Representative wound healing images 0, 6, and 24 h after LTA stimulation. (B) Quantification of wound closure. (C) Representative images of Transwell migration at 6 h and 24 h after LTA stimulation. Migrated cells on the lower surface of the membrane were fixed, stained with crystal violet and images captured under an inverted light microscope. Scale bar, 200 µm. (D) Quantification of migrated cells. Data are presented as means ± SD. ***P<0.001 vs. DMSO; ### P<0.001 vs. DMSO+LTA; † P<0.05, †† P<0.01 and ††† P<0.001 vs. GBD+LTA; ‡‡ P<0.01 vs. ML385+LTA. GBD, glabridin; LTA, lipoteichoic acid; Nrf2, nuclear factor erythroid 2-related factor 2; DMSO, dimethyl sulfoxide.

Article Snippet: Cells were then transfected with either a small interfering (si)RNA targeting Nrf2 [ Nfe2l2 siRNA: 5′-AGCAUUUUAACAUGUUAACAG-3′ (sense) and 5′-GUUAACAUGUUAAAAUGCUAU-3′ (antisense)] or a negative control siRNA [si-Ctrl: 5′-UUCUCCGAACGUGUCACGUTT-3′ (sense) and reverse 5′-ACGUGACACGUUCGGAGAATT-3′ (antisense); cat. no. HY-RS09246; MedChemExpress], using a transfection reagent (cat. no. HY-K2017, MedChemExpress) and 50 nM siRNA diluted in serum-free medium, according to the manufacturer's instructions.

Techniques: Migration, Activation Assay, Membrane, Staining, Light Microscopy

Journal: Molecular Medicine Reports

Article Title: Glabridin attenuates LTA-induced alveolar macrophage migration via activation of the Nrf2/HO-1 pathway

doi: 10.3892/mmr.2025.13758

Figure Lengend Snippet: Schematic of GBD-induced modulation of macrophage migration in vitro . Upon LTA stimulation, MH-S cells increase ROS production, activating Nrf2 and upregulating HO-1. GBD thus modulateS cell migration by increasing Nrf2 nuclear translocation and HO-1 expression. GBD, glabridin; LTA, lipoteichoic acid; ROS, reactive oxygen species; Nrf2, nuclear factor erythroid 2-related factor 2; HO-1, heme oxygenase-1.

Article Snippet: Cells were then transfected with either a small interfering (si)RNA targeting Nrf2 [ Nfe2l2 siRNA: 5′-AGCAUUUUAACAUGUUAACAG-3′ (sense) and 5′-GUUAACAUGUUAAAAUGCUAU-3′ (antisense)] or a negative control siRNA [si-Ctrl: 5′-UUCUCCGAACGUGUCACGUTT-3′ (sense) and reverse 5′-ACGUGACACGUUCGGAGAATT-3′ (antisense); cat. no. HY-RS09246; MedChemExpress], using a transfection reagent (cat. no. HY-K2017, MedChemExpress) and 50 nM siRNA diluted in serum-free medium, according to the manufacturer's instructions.

Techniques: Migration, In Vitro, Translocation Assay, Expressing