Journal: Frontiers in Pharmacology

Article Title: Protective effect of hesperetin against myelosuppression and splenic inflammation via modulating TLR4/MYD88/NF-κB signaling: molecular docking and experimental validation

doi: 10.3389/fphar.2026.1826774

Figure Lengend Snippet: (A) List of ten intersecting genes of TLR–MYD88 pathway interactors that are simultaneously targeted by hesperetin. (B) Lollipop KEGG pathways enrichment chart of the top 10 pathways, color coded and sorted by fold enrichment. Chart was constructed using STRING db (FDR: false discovery rate).

Article Snippet: Then, a citrate buffer solution was added to the tissues and heating was done in a microwave for 24 min. Then, a diluted solution of MYD88 rabbit mAB (1:200, A19082, ABclonal) was added.

Techniques: Construct

Journal: Frontiers in Pharmacology

Article Title: Protective effect of hesperetin against myelosuppression and splenic inflammation via modulating TLR4/MYD88/NF-κB signaling: molecular docking and experimental validation

doi: 10.3389/fphar.2026.1826774

Figure Lengend Snippet: Effect of hesperetin on MYD88 immunohistochemical staining in specimens from mice spleens. (A) Saline control group: Images show a normal MYD88 immunostaining pattern. (B) Vincristine group: show marked increase of MYD88%, correlating with lymphoid depletion. (C) Vincristine + hesperetin group: the MYD88 staining was increased versus the vincristine group. (D1,D2) The hesperetin per se group showed immunostaining similar to the saline group. (E) Column charts for the area of immunostaining, data are mean ± SD. * versus the saline group, # versus vincristine group (P < 0.05).

Article Snippet: Then, a citrate buffer solution was added to the tissues and heating was done in a microwave for 24 min. Then, a diluted solution of MYD88 rabbit mAB (1:200, A19082, ABclonal) was added.

Techniques: Immunohistochemical staining, Staining, Saline, Control, Immunostaining

Journal: Frontiers in Pharmacology

Article Title: Protective effect of hesperetin against myelosuppression and splenic inflammation via modulating TLR4/MYD88/NF-κB signaling: molecular docking and experimental validation

doi: 10.3389/fphar.2026.1826774

Figure Lengend Snippet: Effect of hesperetin on the vincristine induced target biomarkers in the spleen. Bar charts demonstrate inflammatory markers detected by ELISA in spleen supernatants of (A) TLR4, (B) NF-κB, (C) MYD88, and (D) TNF-α, and (E) IL-1β. Data are demonstrated as mean ± SD. * versus the saline group, # versus vincristine group (P < 0.05).

Article Snippet: Then, a citrate buffer solution was added to the tissues and heating was done in a microwave for 24 min. Then, a diluted solution of MYD88 rabbit mAB (1:200, A19082, ABclonal) was added.

Techniques: Enzyme-linked Immunosorbent Assay, Saline

Journal: Frontiers in Pharmacology

Article Title: Protective effect of hesperetin against myelosuppression and splenic inflammation via modulating TLR4/MYD88/NF-κB signaling: molecular docking and experimental validation

doi: 10.3389/fphar.2026.1826774

Figure Lengend Snippet: Correlation using Pearson’s correlation analysis between TLR4 and the other inflammatory markers. The correlation between TLR4 and (A) MYD88, (B) NF-κB, (C) TNF-α and (D) IL-1β are shown.

Article Snippet: Then, a citrate buffer solution was added to the tissues and heating was done in a microwave for 24 min. Then, a diluted solution of MYD88 rabbit mAB (1:200, A19082, ABclonal) was added.

Techniques:

Journal: Frontiers in Microbiology

Article Title: Spermidine protects against ETEC-induced intestinal injury by reshaping the gut microbiota and activating the AhR/IL-22 axis

doi: 10.3389/fmicb.2026.1907051

Figure Lengend Snippet: Spermidine suppresses activation of the TLR4/MyD88/NF-κB signaling pathway in ETEC-infected mice. (A) Representative Western blot images showing the protein expression of TLR4, MyD88, phosphorylated NF-κB p65 (p-p65), total NF-κB p65 (p65), and GAPDH in jejunal tissues. (B–D) Densitometric analyses of TLR4 (B) , MyD88 (C) , and p-p65 (D) protein expression normalized to the corresponding loading controls. Data are presented as mean ± SD. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple-comparison test. ** p < 0.01, *** p < 0.001.

Article Snippet: The primary antibodies used for Western blot were TLR4 (Proteintech, Cat No. 66350-1-Ig, 1:4,000), MyD88 (ABclonal, Cat No. A0980, 1:2,000), p-p65 (ABclonal, Cat No. AP1294, 1:2,000), p65 (ABclonal, Cat No. A19653, 1:5,000), GAPDH (Proteintech, Cat No. 60004-1-Ig, 1:20,000), ZO-1 (Proteintech, Cat No. 21773-1-AP, 1:5,000), occludin (Proteintech, Cat No. 66378-1-Ig, 1:5,000), AhR (Proteintech, Cat No. 67785-1-Ig, 1:2,000), and CYP1A1 (Proteintech, Cat No. 13241-1-AP, 1:1,000).

Techniques: Activation Assay, Infection, Western Blot, Expressing, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Oral Sodium Hyaluronate Reshapes Gut Microbiota Composition and Suppresses the LPS-TLR4/NF-κB Pathway to Exert Neuroprotection in MPTP-Induced Parkinson’s Disease

doi: 10.3390/ijms27156573

Figure Lengend Snippet: Oral SH inhibited the TLR4/MyD88/NF-κB signaling pathways both in the SN and the colon of MPTP-induced mice. ( A ) Schematic representation of the signaling pathway. ( B ) Representative WB bands of TLR4, MyD88, and NF-κB in the colon. ( C ) Representative WB bands of TLR4, MyD88, and NF-κB in the midbrain containing the SN. ( D – F ) The density analysis results of TLR4, MyD88, and NF-κB in the colon. ( G – I ) The density analysis results of TLR4, MyD88, and NF-κB in the midbrain containing the SN. ( n = 3 for each group. Data are presented as mean ± SD. *** p < 0.001 versus the control group; ### p < 0.001 versus the MPTP group.).

Article Snippet: Membranes were incubated at 4 °C overnight with the following primary antibodies: rabbit anti-β-actin antibody (1:50,000, AC026, Abclonal, Wuhan, China), mouse anti-TH antibody (1:1000, A0028, Abclonal, Wuhan, China), rabbit anti-TNF-α antibody (1:1000, A20851, Abclonal, Wuhan, China), rabbit anti-IL-1β antibody (1:1000, A11369, Abclonal, Wuhan, China), rabbit anti-IL-6 antibody (1:1000, A0286, Abclonal, Wuhan, China), goat anti-COX-2 antibody (1:1000, A1253, Abclonal, Wuhan, China), mouse anti-TLR4 antibody (1:1000, A5258, Abclonal, Wuhan, China), rabbit anti-MyD88 antibody (1:400, A0980, Abclonal, Wuhan, China), rabbit anti-IκB-α antibody (1:1000, A24742, Abclonal, Wuhan, China), rabbit anti-ZO-1 antibody (1:1000, A0659, Abclonal, Wuhan, China), mouse anti-claudin-1 antibody (1:1000, Abclonal, Wuhan, China), rabbit anti-occludin antibody (1:1000, Abclonal, China), and mouse anti-claudin-5 antibody (1:1000, A10207 , Abclonal, Wuhan, China).

Techniques: Protein-Protein interactions, Control

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Electroacupuncture Attenuates Cerebral Ischemia–Reperfusion Injury via Vagus Nerve–α7 Nicotinic Acetylcholine Receptor–Dependent Microglial Polarization

doi: 10.1161/JAHA.126.049344

Figure Lengend Snippet: A , qRT‐PCR analysis of α7nAChR mRNA levels in the ischemic penumbra at 1, 3, and 7 d after MCAO (n=3 per group). B , C , Representative Western blots and quantification of α7nAChR protein levels in the penumbra at the indicated time points (n=3 per group). D , E , Representative immunofluorescence images showing α7nAChR (red) and Iba1 (green) in the penumbra at d 3 post MCAO, and quantification of their fluorescence colocalization intensity (n=3 per group; scale bar, 100 μm). F , G , Representative immunofluorescence images showing α7nAChR (red) and NeuN (green) in the penumbra, and quantification of co‐localization intensity (n=3 per group; scale bar, 100 μm). H , I , Neurological deficit scores (modified Garcia, Petullo) at d 3 after MCAO following α‐BGT administration (n=6 per group). J , K , TTC staining and quantitative analysis of infarct volume (n=6 per group). In ( A ) and ( C ), values in the Sham group were normalized to 1 as a reference to present relative expression changes. In ( H , I , and K ), the identical values in the Sham group reflect normal physiological baselines by design. α7nAChR indicates α7 nicotinic acetylcholine receptor; α‐BGT, α‐bungarotoxin; EA, electroacupuncture; Iba‐1, ionized calcium binding adaptor molecule 1; MCAO/R, middle cerebral artery occlusion/reperfusion; NeuN, neuronal nuclei; qRT‐PCR, quantitative real‐time polymerase chain reaction; and TTC, 2,3,5‐triphenyltetrazolium chloride.

Article Snippet: The following primary antibodies were used: Arg‐1 (arginase 1; 1:30 000; Proteintech, #16001‐1‐AP), TGF‐β1 (transforming growth factor beta 1; 1:1000; Abclonal, #A18692), iNOS (inducible nitric oxide synthase; 1:1500; Affinity, #AF0199), TNF‐α (tumor necrosis factor alpha; 1:1500; Wanleibio, #WL01581), TLR4 (1:1000; Cell Signaling Technology, #14358T), MyD88 (1:500; Abclonal, #A0980), α7nAChR (1:500; Abclonal, #A1588), β‐tubulin (1:100000; Proteintech, #80713‐1‐RR), p‐IκBα (phospho‐IkappaB alpha; 1:800; Abclonal, #AP0999), IκBα (1:1000; Abclonal, #A11168), NF‐κB p65 (1:1000; Cell Signaling Technology, #8242S), histone (Proteintech, #17168‐1‐AP), and β‐actin (1:1000; Wanleibio, #WL01372).

Techniques: Quantitative RT-PCR, Western Blot, Immunofluorescence, Fluorescence, Modification, Staining, Expressing, Binding Assay, Real-time Polymerase Chain Reaction

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Electroacupuncture Attenuates Cerebral Ischemia–Reperfusion Injury via Vagus Nerve–α7 Nicotinic Acetylcholine Receptor–Dependent Microglial Polarization

doi: 10.1161/JAHA.126.049344

Figure Lengend Snippet: Tissues were collected from the ischemic penumbra at d 3 after MCAO. A through C , Representative Western blots and quantification of anti‐inflammatory markers Arg‐1 and TGF‐β1 (n=3 per group). D through F , Representative Western blots and quantification of proinflammatory markers iNOS and TNF‐α (n=3 per group). G through L , Representative Western blots and quantification of Arg‐1, TGF‐β1, iNOS, and TNF‐α following α7nAChR antagonism with α‐BGT (n=3 per group). M , N , Representative immunofluorescence images showing colocalization of Arg‐1 (red) and iNOS (red) with the microglial marker Iba‐1 (green) in the penumbra (scale bar, 100 μm). O through R , ELISA analysis of anti‐inflammatory markers (CD206, IL‐10) and proinflammatory markers (CD86, IL‐1β) (n=6 per group). α7nAChR indicates α7 nicotinic acetylcholine receptor; α‐BGT, α‐bungarotoxin; Arg‐1, arginase 1; EA, electroacupuncture; Iba‐1, ionized calcium binding adaptor molecule 1; IL, interleukin; iNOS, inducible nitric oxide synthase; MCAO/R, middle cerebral artery occlusion/reperfusion; TGF‐β1, transforming growth factor beta 1; and TNF‐α, tumor necrosis factor alpha.

Article Snippet: The following primary antibodies were used: Arg‐1 (arginase 1; 1:30 000; Proteintech, #16001‐1‐AP), TGF‐β1 (transforming growth factor beta 1; 1:1000; Abclonal, #A18692), iNOS (inducible nitric oxide synthase; 1:1500; Affinity, #AF0199), TNF‐α (tumor necrosis factor alpha; 1:1500; Wanleibio, #WL01581), TLR4 (1:1000; Cell Signaling Technology, #14358T), MyD88 (1:500; Abclonal, #A0980), α7nAChR (1:500; Abclonal, #A1588), β‐tubulin (1:100000; Proteintech, #80713‐1‐RR), p‐IκBα (phospho‐IkappaB alpha; 1:800; Abclonal, #AP0999), IκBα (1:1000; Abclonal, #A11168), NF‐κB p65 (1:1000; Cell Signaling Technology, #8242S), histone (Proteintech, #17168‐1‐AP), and β‐actin (1:1000; Wanleibio, #WL01372).

Techniques: Western Blot, Immunofluorescence, Marker, Enzyme-linked Immunosorbent Assay, Binding Assay

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Electroacupuncture Attenuates Cerebral Ischemia–Reperfusion Injury via Vagus Nerve–α7 Nicotinic Acetylcholine Receptor–Dependent Microglial Polarization

doi: 10.1161/JAHA.126.049344

Figure Lengend Snippet: A , Representative Western blots of total TLR4, MyD88, p‐IκBα, IκBα, and nuclear NF‐κB p65 in the ischemic penumbra at day 3 after MCAO. B through E , Quantitative analysis of protein levels for total TLR4, MyD88, p‐IκBα/IκBα ratio, and nuclear NF‐κB p65 (n=3 per group). F , G , Representative immunofluorescence images showing TLR4 and NF‐κB p65 expression in the penumbra (scale bar, 100 μm). α7nAChR indicates α7 nicotinic acetylcholine receptor; α‐BGT, α‐bungarotoxin; EA, electroacupuncture; MCAO/R, middle cerebral artery occlusion/reperfusion; MyD88, myeloid differentiation primary response 88; NF‐κB, nuclear factor kappa B; p‐IκBα, phospho‐IkappaB alpha; and TLR4, toll‐like receptor 4.

Article Snippet: The following primary antibodies were used: Arg‐1 (arginase 1; 1:30 000; Proteintech, #16001‐1‐AP), TGF‐β1 (transforming growth factor beta 1; 1:1000; Abclonal, #A18692), iNOS (inducible nitric oxide synthase; 1:1500; Affinity, #AF0199), TNF‐α (tumor necrosis factor alpha; 1:1500; Wanleibio, #WL01581), TLR4 (1:1000; Cell Signaling Technology, #14358T), MyD88 (1:500; Abclonal, #A0980), α7nAChR (1:500; Abclonal, #A1588), β‐tubulin (1:100000; Proteintech, #80713‐1‐RR), p‐IκBα (phospho‐IkappaB alpha; 1:800; Abclonal, #AP0999), IκBα (1:1000; Abclonal, #A11168), NF‐κB p65 (1:1000; Cell Signaling Technology, #8242S), histone (Proteintech, #17168‐1‐AP), and β‐actin (1:1000; Wanleibio, #WL01372).

Techniques: Western Blot, Immunofluorescence, Expressing

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Electroacupuncture Attenuates Cerebral Ischemia–Reperfusion Injury via Vagus Nerve–α7 Nicotinic Acetylcholine Receptor–Dependent Microglial Polarization

doi: 10.1161/JAHA.126.049344

Figure Lengend Snippet: A and B , Representative immunofluorescence images showing c‐Fos (red) and ChAT (green) in the DMV at d 3 post MCAO, and quantitative analysis of their co‐localization (n=3 per group; scale bar, 200 μm). C and D , Representative Western blots and quantification of α7nAChR protein levels in the penumbra following SDV (n=3 per group). E , F , Neurological deficit scores (modified Garcia, Petullo) at d 3 after MCAO (n=6 per group). G , H , TTC staining and quantitative analysis of infarct volume (n=6 per group). The identical values in the Sham group reflect normal physiological baselines by design. α7nAChR indicates α7 nicotinic acetylcholine receptor; α‐BGT, α‐bungarotoxin; ChAT, choline acetyltransferase; EA, electroacupuncture; MCAO/R, middle cerebral artery occlusion/reperfusion; SDV, subdiaphragmatic vagotomy; and TTC, 2,3,5‐triphenyltetrazolium chloride.

Article Snippet: The following primary antibodies were used: Arg‐1 (arginase 1; 1:30 000; Proteintech, #16001‐1‐AP), TGF‐β1 (transforming growth factor beta 1; 1:1000; Abclonal, #A18692), iNOS (inducible nitric oxide synthase; 1:1500; Affinity, #AF0199), TNF‐α (tumor necrosis factor alpha; 1:1500; Wanleibio, #WL01581), TLR4 (1:1000; Cell Signaling Technology, #14358T), MyD88 (1:500; Abclonal, #A0980), α7nAChR (1:500; Abclonal, #A1588), β‐tubulin (1:100000; Proteintech, #80713‐1‐RR), p‐IκBα (phospho‐IkappaB alpha; 1:800; Abclonal, #AP0999), IκBα (1:1000; Abclonal, #A11168), NF‐κB p65 (1:1000; Cell Signaling Technology, #8242S), histone (Proteintech, #17168‐1‐AP), and β‐actin (1:1000; Wanleibio, #WL01372).

Techniques: Immunofluorescence, Western Blot, Modification, Staining