Journal: Scientific Reports

Article Title: Lactobacillus yoelii Lac-2 mediates the mechanism of MAPK signaling pathway involved in the regulation of intestinal barrier and pathogen translocation by Zonulin expression

doi: 10.1038/s41598-026-55430-y

Figure Lengend Snippet: The mRNA expression levels of mucin and TJ proteins in each treatment group after the addition of MAPK activator to Zonulin overexpressing cell lines. ( A ) Relative expression of MUC1 mRNA; ( B ) Relative expression of MUC2 mRNA; ( C ) Relative expression of ZO-1 mRNA; ( D ) Relative mRNA expression of Occludin; ( E ) Relative expression of Claudin 1 mRNA; ( F ) Relative expression of Zonulin mRNA. * p < 0.05, ** p < 0.01.

Article Snippet: Primary antibodies included p38 rabbit polyclonal antibody (A0027, Abclonal, Wuhan, China), ERK rabbit polyclonal antibody (AP1120, Abclonal, Wuhan, China), JNK rabbit polyclonal antibody (A18678, Abclonal, Wuhan, China), zonulin rabbit polyclonal antibody (A1571, Abclonal, Wuhan, China), β-Actin rabbit monoclonal antibody (highly sensitive) (AC026, Abclonal, Wuhan, China), MUC1 rabbit polyclonal antibody (A21726, Abclonal, Wuhan, China), MUC2 rabbit polyclonal antibody (A14659, Abclonal, Wuhan, China), ZO-1 rabbit polyclonal antibody (A0659, Abclonal, Wuhan, China), occludin rabbit polyclonal antibody (A12621, Abclonal, Wuhan, China), claudin 1 rabbit polyclonal antibody (A21770, Abclonal, Wuhan, China), and HRP-conjugated sheep anti-rabbit IgG (H + L) (A0208, Abclonal, Wuhan, China).

Techniques: Expressing

Journal: Scientific Reports

Article Title: Lactobacillus yoelii Lac-2 mediates the mechanism of MAPK signaling pathway involved in the regulation of intestinal barrier and pathogen translocation by Zonulin expression

doi: 10.1038/s41598-026-55430-y

Figure Lengend Snippet: The mRNA expression levels of mucin and TJ proteins in each treatment group after the addition of MAPK blocker to the Zonulin up-regulation cell line. ( A ) Relative expression of MUC1 mRNA; ( B ) Relative expression of MUC2 mRNA; ( C ) Relative expression of ZO-1 mRNA; ( D ) Relative mRNA expression of Occludin; ( E ) Relative expression of Claudin 1 mRNA; ( F ) Relative expression of Zonulin mRNA. * p < 0.05, ** p < 0.01.

Article Snippet: Primary antibodies included p38 rabbit polyclonal antibody (A0027, Abclonal, Wuhan, China), ERK rabbit polyclonal antibody (AP1120, Abclonal, Wuhan, China), JNK rabbit polyclonal antibody (A18678, Abclonal, Wuhan, China), zonulin rabbit polyclonal antibody (A1571, Abclonal, Wuhan, China), β-Actin rabbit monoclonal antibody (highly sensitive) (AC026, Abclonal, Wuhan, China), MUC1 rabbit polyclonal antibody (A21726, Abclonal, Wuhan, China), MUC2 rabbit polyclonal antibody (A14659, Abclonal, Wuhan, China), ZO-1 rabbit polyclonal antibody (A0659, Abclonal, Wuhan, China), occludin rabbit polyclonal antibody (A12621, Abclonal, Wuhan, China), claudin 1 rabbit polyclonal antibody (A21770, Abclonal, Wuhan, China), and HRP-conjugated sheep anti-rabbit IgG (H + L) (A0208, Abclonal, Wuhan, China).

Techniques: Expressing

Journal: Scientific Reports

Article Title: Lactobacillus yoelii Lac-2 mediates the mechanism of MAPK signaling pathway involved in the regulation of intestinal barrier and pathogen translocation by Zonulin expression

doi: 10.1038/s41598-026-55430-y

Figure Lengend Snippet: The mRNA expression levels of mucin and TJ protein in each treatment group were reduced by Zonulin knockdown cell line after MAPK activator was added. ( A ) Relative expression of MUC1 mRNA; ( B ) Relative expression of MUC2 mRNA; ( C ) Relative expression of ZO-1 mRNA; ( D ) Relative mRNA expression of Occludin; ( E ) Relative expression of Claudin 1 mRNA; ( F ) Relative expression of Zonulin mRNA. * p < 0.05, ** p < 0.01.

Article Snippet: Primary antibodies included p38 rabbit polyclonal antibody (A0027, Abclonal, Wuhan, China), ERK rabbit polyclonal antibody (AP1120, Abclonal, Wuhan, China), JNK rabbit polyclonal antibody (A18678, Abclonal, Wuhan, China), zonulin rabbit polyclonal antibody (A1571, Abclonal, Wuhan, China), β-Actin rabbit monoclonal antibody (highly sensitive) (AC026, Abclonal, Wuhan, China), MUC1 rabbit polyclonal antibody (A21726, Abclonal, Wuhan, China), MUC2 rabbit polyclonal antibody (A14659, Abclonal, Wuhan, China), ZO-1 rabbit polyclonal antibody (A0659, Abclonal, Wuhan, China), occludin rabbit polyclonal antibody (A12621, Abclonal, Wuhan, China), claudin 1 rabbit polyclonal antibody (A21770, Abclonal, Wuhan, China), and HRP-conjugated sheep anti-rabbit IgG (H + L) (A0208, Abclonal, Wuhan, China).

Techniques: Expressing, Knockdown

Journal: Scientific Reports

Article Title: Lactobacillus yoelii Lac-2 mediates the mechanism of MAPK signaling pathway involved in the regulation of intestinal barrier and pathogen translocation by Zonulin expression

doi: 10.1038/s41598-026-55430-y

Figure Lengend Snippet: The mRNA expression levels of mucin and TJ proteins in each treatment group after Zonulin knockdown cell line was supplemented with MAPK blocker. ( A ) Relative expression of MUC1 mRNA; ( B ) Relative expression of MUC2 mRNA; ( C ) Relative expression of ZO-1 mRNA; ( D ) Relative mRNA expression of Occludin; ( E ) Relative expression of Claudin 1 mRNA; ( F ) Relative expression of Zonulin mRNA. * p < 0.05, ** p < 0.01.

Article Snippet: Primary antibodies included p38 rabbit polyclonal antibody (A0027, Abclonal, Wuhan, China), ERK rabbit polyclonal antibody (AP1120, Abclonal, Wuhan, China), JNK rabbit polyclonal antibody (A18678, Abclonal, Wuhan, China), zonulin rabbit polyclonal antibody (A1571, Abclonal, Wuhan, China), β-Actin rabbit monoclonal antibody (highly sensitive) (AC026, Abclonal, Wuhan, China), MUC1 rabbit polyclonal antibody (A21726, Abclonal, Wuhan, China), MUC2 rabbit polyclonal antibody (A14659, Abclonal, Wuhan, China), ZO-1 rabbit polyclonal antibody (A0659, Abclonal, Wuhan, China), occludin rabbit polyclonal antibody (A12621, Abclonal, Wuhan, China), claudin 1 rabbit polyclonal antibody (A21770, Abclonal, Wuhan, China), and HRP-conjugated sheep anti-rabbit IgG (H + L) (A0208, Abclonal, Wuhan, China).

Techniques: Expressing, Knockdown

Journal: Scientific Reports

Article Title: Lactobacillus yoelii Lac-2 mediates the mechanism of MAPK signaling pathway involved in the regulation of intestinal barrier and pathogen translocation by Zonulin expression

doi: 10.1038/s41598-026-55430-y

Figure Lengend Snippet: Zonulin up-regulation and Zonulin knockdown cell lines were treated with MAPK activator and MAPK blocker mRNA expression levels. ( A ) Relative expression of MUC1 mRNA; ( B ) Relative expression of MUC2 mRNA; ( C ) Relative expression of ZO-1 mRNA; ( D ) Relative mRNA expression of Occludin; ( E ) Relative expression of Claudin 1 mRNA; ( F ) Relative expression of Zonulin mRNA. * p < 0.05, ** p < 0.01.

Article Snippet: Primary antibodies included p38 rabbit polyclonal antibody (A0027, Abclonal, Wuhan, China), ERK rabbit polyclonal antibody (AP1120, Abclonal, Wuhan, China), JNK rabbit polyclonal antibody (A18678, Abclonal, Wuhan, China), zonulin rabbit polyclonal antibody (A1571, Abclonal, Wuhan, China), β-Actin rabbit monoclonal antibody (highly sensitive) (AC026, Abclonal, Wuhan, China), MUC1 rabbit polyclonal antibody (A21726, Abclonal, Wuhan, China), MUC2 rabbit polyclonal antibody (A14659, Abclonal, Wuhan, China), ZO-1 rabbit polyclonal antibody (A0659, Abclonal, Wuhan, China), occludin rabbit polyclonal antibody (A12621, Abclonal, Wuhan, China), claudin 1 rabbit polyclonal antibody (A21770, Abclonal, Wuhan, China), and HRP-conjugated sheep anti-rabbit IgG (H + L) (A0208, Abclonal, Wuhan, China).

Techniques: Knockdown, Expressing

Journal: Scientific Reports

Article Title: Lactobacillus yoelii Lac-2 mediates the mechanism of MAPK signaling pathway involved in the regulation of intestinal barrier and pathogen translocation by Zonulin expression

doi: 10.1038/s41598-026-55430-y

Figure Lengend Snippet: The expression levels of mucin and TJ protein in each treatment group after the addition of MAPK activator to Zonulin up-regulation cell line. ( A ) Gray value detection results of mucin WB; ( B ) Gray value of TJ protein WB; ( C ) Relative β-actin expression level of MUC1; ( D ) Relative β-actin expression level of MUC2; ( E ) Relative β-actin expression level of ZO-1; ( F ) Relative β-actin expression level of Occludin; ( G ) Claudin 1 relative β-actin expression level; ( H ) Relative β-actin expression levels of Zonulin. * p < 0.05, ** p < 0.01.

Article Snippet: Primary antibodies included p38 rabbit polyclonal antibody (A0027, Abclonal, Wuhan, China), ERK rabbit polyclonal antibody (AP1120, Abclonal, Wuhan, China), JNK rabbit polyclonal antibody (A18678, Abclonal, Wuhan, China), zonulin rabbit polyclonal antibody (A1571, Abclonal, Wuhan, China), β-Actin rabbit monoclonal antibody (highly sensitive) (AC026, Abclonal, Wuhan, China), MUC1 rabbit polyclonal antibody (A21726, Abclonal, Wuhan, China), MUC2 rabbit polyclonal antibody (A14659, Abclonal, Wuhan, China), ZO-1 rabbit polyclonal antibody (A0659, Abclonal, Wuhan, China), occludin rabbit polyclonal antibody (A12621, Abclonal, Wuhan, China), claudin 1 rabbit polyclonal antibody (A21770, Abclonal, Wuhan, China), and HRP-conjugated sheep anti-rabbit IgG (H + L) (A0208, Abclonal, Wuhan, China).

Techniques: Expressing

Journal: Scientific Reports

Article Title: Lactobacillus yoelii Lac-2 mediates the mechanism of MAPK signaling pathway involved in the regulation of intestinal barrier and pathogen translocation by Zonulin expression

doi: 10.1038/s41598-026-55430-y

Figure Lengend Snippet: Expression levels of mucin and TJ protein in each treatment group after the addition of MAPK blocker to Zonulin up-regulation cell line. ( A ) Gray value detection results of mucin WB; ( B ) Gray value of TJ protein WB; ( C ) Relative β-actin expression level of MUC1; ( D ) Relative β-actin expression level of MUC2; ( E ) Relative β-actin expression level of ZO-1; ( F ) Relative β-actin expression level of Occludin; ( G ) Claudin 1 relative β-actin expression level; ( H ) Relative β-actin expression levels of Zonulin. * p < 0.05, ** p < 0.01.

Article Snippet: Primary antibodies included p38 rabbit polyclonal antibody (A0027, Abclonal, Wuhan, China), ERK rabbit polyclonal antibody (AP1120, Abclonal, Wuhan, China), JNK rabbit polyclonal antibody (A18678, Abclonal, Wuhan, China), zonulin rabbit polyclonal antibody (A1571, Abclonal, Wuhan, China), β-Actin rabbit monoclonal antibody (highly sensitive) (AC026, Abclonal, Wuhan, China), MUC1 rabbit polyclonal antibody (A21726, Abclonal, Wuhan, China), MUC2 rabbit polyclonal antibody (A14659, Abclonal, Wuhan, China), ZO-1 rabbit polyclonal antibody (A0659, Abclonal, Wuhan, China), occludin rabbit polyclonal antibody (A12621, Abclonal, Wuhan, China), claudin 1 rabbit polyclonal antibody (A21770, Abclonal, Wuhan, China), and HRP-conjugated sheep anti-rabbit IgG (H + L) (A0208, Abclonal, Wuhan, China).

Techniques: Expressing

Journal: Scientific Reports

Article Title: Lactobacillus yoelii Lac-2 mediates the mechanism of MAPK signaling pathway involved in the regulation of intestinal barrier and pathogen translocation by Zonulin expression

doi: 10.1038/s41598-026-55430-y

Figure Lengend Snippet: The expression levels of mucin and TJ protein in each treatment group after Zonulin knockdown cell line was supplemented with MAPK activator. ( A ) Gray value detection results of mucin WB; ( B ) Gray value of TJ protein WB; ( C ) Relative β-actin expression level of MUC1; ( D ) Relative β-actin expression level of MUC2; ( E ) Relative β-actin expression level of ZO-1; ( F ) Relative β-actin expression level of Occludin; ( G ) Claudin 1 relative β-actin expression level; ( H ) Relative β-actin expression levels of Zonulin. * p < 0.05, ** p < 0.01.

Article Snippet: Primary antibodies included p38 rabbit polyclonal antibody (A0027, Abclonal, Wuhan, China), ERK rabbit polyclonal antibody (AP1120, Abclonal, Wuhan, China), JNK rabbit polyclonal antibody (A18678, Abclonal, Wuhan, China), zonulin rabbit polyclonal antibody (A1571, Abclonal, Wuhan, China), β-Actin rabbit monoclonal antibody (highly sensitive) (AC026, Abclonal, Wuhan, China), MUC1 rabbit polyclonal antibody (A21726, Abclonal, Wuhan, China), MUC2 rabbit polyclonal antibody (A14659, Abclonal, Wuhan, China), ZO-1 rabbit polyclonal antibody (A0659, Abclonal, Wuhan, China), occludin rabbit polyclonal antibody (A12621, Abclonal, Wuhan, China), claudin 1 rabbit polyclonal antibody (A21770, Abclonal, Wuhan, China), and HRP-conjugated sheep anti-rabbit IgG (H + L) (A0208, Abclonal, Wuhan, China).

Techniques: Expressing, Knockdown

Journal: Scientific Reports

Article Title: Lactobacillus yoelii Lac-2 mediates the mechanism of MAPK signaling pathway involved in the regulation of intestinal barrier and pathogen translocation by Zonulin expression

doi: 10.1038/s41598-026-55430-y

Figure Lengend Snippet: The expression levels of mucin and TJ protein in each treatment group after Zonulin knockdown cell line was supplemented with MAPK blocker. ( A ) Gray value detection results of mucin WB; ( B ) Gray value of TJ protein WB; ( C ) Relative β-actin expression level of MUC1; ( D ) Relative β-actin expression level of MUC2; ( E ) Relative β-actin expression level of ZO-1; ( F ) Relative β-actin expression level of Occludin; ( G ) Claudin 1 relative β-actin expression level; ( H ) Relative β-actin expression levels of Zonulin. * p < 0.05, ** p < 0.01.

Article Snippet: Primary antibodies included p38 rabbit polyclonal antibody (A0027, Abclonal, Wuhan, China), ERK rabbit polyclonal antibody (AP1120, Abclonal, Wuhan, China), JNK rabbit polyclonal antibody (A18678, Abclonal, Wuhan, China), zonulin rabbit polyclonal antibody (A1571, Abclonal, Wuhan, China), β-Actin rabbit monoclonal antibody (highly sensitive) (AC026, Abclonal, Wuhan, China), MUC1 rabbit polyclonal antibody (A21726, Abclonal, Wuhan, China), MUC2 rabbit polyclonal antibody (A14659, Abclonal, Wuhan, China), ZO-1 rabbit polyclonal antibody (A0659, Abclonal, Wuhan, China), occludin rabbit polyclonal antibody (A12621, Abclonal, Wuhan, China), claudin 1 rabbit polyclonal antibody (A21770, Abclonal, Wuhan, China), and HRP-conjugated sheep anti-rabbit IgG (H + L) (A0208, Abclonal, Wuhan, China).

Techniques: Expressing, Knockdown

Journal: Scientific Reports

Article Title: Lactobacillus yoelii Lac-2 mediates the mechanism of MAPK signaling pathway involved in the regulation of intestinal barrier and pathogen translocation by Zonulin expression

doi: 10.1038/s41598-026-55430-y

Figure Lengend Snippet: Zonulin up-regulation and Zonulin knockdown cell lines were treated with MAPK activator and MAPK blocker. ( A ) Gray value detection results of mucin WB; ( B ) Gray value of TJ protein WB; ( C ) Relative β-actin expression level of MUC1; ( D ) Relative β-actin expression level of MUC2; ( E ) Relative β-actin expression level of ZO-1; ( F ) Relative β-actin expression level of Occludin; ( G ) Claudin 1 relative β-actin expression level; ( H ) Relative β-actin expression levels of Zonulin. * p < 0.05, ** p < 0.01.

Article Snippet: Primary antibodies included p38 rabbit polyclonal antibody (A0027, Abclonal, Wuhan, China), ERK rabbit polyclonal antibody (AP1120, Abclonal, Wuhan, China), JNK rabbit polyclonal antibody (A18678, Abclonal, Wuhan, China), zonulin rabbit polyclonal antibody (A1571, Abclonal, Wuhan, China), β-Actin rabbit monoclonal antibody (highly sensitive) (AC026, Abclonal, Wuhan, China), MUC1 rabbit polyclonal antibody (A21726, Abclonal, Wuhan, China), MUC2 rabbit polyclonal antibody (A14659, Abclonal, Wuhan, China), ZO-1 rabbit polyclonal antibody (A0659, Abclonal, Wuhan, China), occludin rabbit polyclonal antibody (A12621, Abclonal, Wuhan, China), claudin 1 rabbit polyclonal antibody (A21770, Abclonal, Wuhan, China), and HRP-conjugated sheep anti-rabbit IgG (H + L) (A0208, Abclonal, Wuhan, China).

Techniques: Knockdown, Expressing

Journal: Frontiers in Toxicology

Article Title: Oridonin attenuates diclofenac-induced organ damage by suppressing endoplasmic reticulum stress and inflammasome activation

doi: 10.3389/ftox.2026.1821778

Figure Lengend Snippet: Influence of oridonin on the alterations in stomach occludin expression induced by diclofenac. (A) Representative microscopic images of glandular stomach sections immunostained for occludin. (B) Histogram quantifies the percentage of the stained area for occludin expression. Normal control group showing strong positive brown staining in many glandular cells (black arrows), Diclofenac group revealing weak positive staining. Oridonin 10 mg/kg group showing moderate positive staining, and Oridonin 20 mg/kg group showing strong positive staining. IHC was counterstained with Mayer’s hematoxylin; magnifications are at ×100 (100 μm bar) and 400X (25 μm bar). Data are expressed as mean ± SD; n = 6. Rats received oridonin (10 mg/kg and 20 mg/kg) for 7 days by gastric gavage followed by diclofenac (100 mg/kg, single IP dose) on 8 th day; **** denotes p-value <0.0001; This was determined using one-way ANOVA followed by Tukey multiple comparison post hoc tests.

Article Snippet: Occludin: Anti-Occludin Rabbit Polyclonal Antibody (Cat# A2601; ABclonal, Woburn, MA, USA).

Techniques: Expressing, Staining, Control, Comparison

Journal: Neural Regeneration Research

Article Title: Chitosan alleviates symptoms of Parkinson’s disease by reducing acetate levels, which decreases inflammation and promotes repair of the intestinal barrier and blood–brain barrier

doi: 10.4103/NRR.NRR-D-23-01511

Figure Lengend Snippet: Chitosan treatment repairs damaged intestinal and blood–brain barriers in an MPTP-induced mouse model of PD. (A) Chitosan administration significantly increased ZO-1 and occludin expression levels, as detected by western blot. GAPDH was used as a loading control. (B) Chitosan treatment significantly increased the fluorescence intensity of ZO-1 (green, Alexa Fluor 488) and occludin (red, Alexa Fluor 594) in mouse colon tissue compared with the MPTP-induced PD group, and the fluorescence intensities of ZO-1 and occludin in PD group were lower than those in the control group. Scale bars: 10 μm. (C) Compared with MPTP-induced PD mice, chitosan treatment significantly decreased serum FITC-dextran levels, which are a measure of intestinal barrier integrity. (D) EB was used to monitor BBB permeability, and results were normalized to the control group. Chitosan treatment significantly reduced BBB damage. (E) EB measured by fluorescence microscopy imaging. Chitosan significantly restored BBB compared with MPTP-induced PD mice. Scale bars: 500 μm (upper) and 50 μm (lower). (F) EB of brain in mice measured by microplate reader. Chitosan treatment significantly decreased EB content compared with MPTP mice. All data are presented as the mean ± SD ( n = 3/group). * P < 0.05 (one-way analysis of variance followed by Tukey’s multiple comparisons test). BBB: Blood–brain barrier; DAPI: 4′,6-diamidino-2-phenylindole; EB: Evans blue; FITC-Dextran: Fluorescein isothiocyanate dextran; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; PD: Parkinson’s disease; ZO-1: Zonula occludens-1.

Article Snippet: The primary antibodies used were as follows: rabbit anti-glyceraldehyde-3-phosphate dehydrogenase polyclonal antibody (GAPDH; 1:10,000, Proteintech, Wuhan, Hubei, China, Cat# 10494-1-AP, RRID: AB_2263076), rabbit anti-TH polyclonal antibody (1:5000, Proteintech, Cat# 25859-1-AP, RRID: AB_2716568), rabbit anti-zonula occludens-1 polyclonal antibody (ZO-1; 1:5000, Proteintech, Cat# 21773-1-AP, RRID: AB_10733242), rabbit anti-occludin polyclonal antibody (1:15,000, Proteintech, Cat# 27260-1-AP, RRID: AB_2880820), rabbit anti-AMPKα polyclonal antibody (1:1000, Cell Signaling Technology, Danvers, Massachusetts, USA, Cat# 2532, RRID: AB_330331), rabbit anti-phospho-AMPKα monoclonal antibody (1:1000, Cell Signaling Technology, Cat# 2535, RRID: AB_331250), and rabbit anti-PPARD polyclonal antibody (1:1000, Abcam, Cambridge, UK, Cat# ab23673, RRID: AB_2165902).

Techniques: Expressing, Western Blot, Control, Fluorescence, Permeability, Microscopy, Imaging

Journal: Neural Regeneration Research

Article Title: Chitosan alleviates symptoms of Parkinson’s disease by reducing acetate levels, which decreases inflammation and promotes repair of the intestinal barrier and blood–brain barrier

doi: 10.4103/NRR.NRR-D-23-01511

Figure Lengend Snippet: Acetate reverses chitosan-mediated repair of the intestinal barrier, increased inflammation in the colon, plasma, and SN, and promotes microglia activation in an MPTP-induced mouse model of PD. (A) Colon length ( n = 5/group). (B) ZO-1 and occludin expression, as assessed by western blot ( n = 3/group). All target proteins were normalized to the reference protein GAPDH. Compared with the chitosan group, acetate supplementation reduced ZO-1 and occludin expression levels. (C) Immunofluorescence staining for ZO-1 (green, Alexa Fluor 488) and occludin (red, Alexa Fluor 594) in mouse colon tissue ( n = 3/group). The immunofluorescence results were consistent with the western blot results. Scale bars: 10 μm. (D) The relative mRNA levels of IL-1β, IL-6, IL-8, IL-10, TNF-α, and iNOS in mouse colon tissue were measured by QPCR ( n = 3/group). Compared with the chitosan group, acetate supplementation resulted in an increase in IL-1β, IL-6, IL-10, TNF-α, and iNOS expression levels in the colon. The data shown in B-D were normalized to the control group. (E) The expression levels of inflammatory cytokines, including IL-1β, IL-6, IL-10, and TNF-α, in mouse plasma were measured by ELISA ( n = 5/group). Compared with the chitosan group, IL-1β and TNF-α levels were significantly increased in the plasma of the acetate group, while IL-10 expression was significantly decreased. (F) The mRNA levels of IL-1β, IL-6, IL-8, IL-10, TNF-α, and iNOS (normalized to the control group) in mouse SN tissue were determined via QPCR ( n = 3/group). Treatment with acetate enhanced TNF-α expression and decreased IL-6 and IL-10 expression in the SN. (G) Representative images of immunofluorescence staining for Iba1 (green, Alexa Fluor 488) and TH (red, Alexa Fluor 594) in the SN ( n = 3/group). The chitosan group exhibited fewer microglia than the MPTP group, while the chitosan + acetate group exhibited more microglia than the chitosan-only group. Scale bars: 50 μm. All data are presented as the mean ± SD. All experiments were repeated at least three times. * P < 0.05 (one-way analysis of variance followed by Tukey’s multiple comparisons test [A–C, G] or unpaired t -test [D–F]). DAPI: 4′,6-Diamidino-2-phenylindole; ELISA: enzyme-linked immunosorbent assay; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; Iba1: ionized calcium-binding adapter molecule 1; IL-1β: interleukin-1 beta; IL-6: interleukin-6; IL-8: interleukin-8; IL-10: interleukin-10; iNOS: inductible nitric oxide synthase; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; n.s.: no significance; NaA: sodium acetate; PD: Parkinson’s disease; QPCR: quantitative polymerase chain reaction; SN: substantia nigra; TH: tyrosine hydroxylase; TNF-α: tumor necrosis factor alpha; ZO-1: Zonula occludens-1.

Article Snippet: The primary antibodies used were as follows: rabbit anti-glyceraldehyde-3-phosphate dehydrogenase polyclonal antibody (GAPDH; 1:10,000, Proteintech, Wuhan, Hubei, China, Cat# 10494-1-AP, RRID: AB_2263076), rabbit anti-TH polyclonal antibody (1:5000, Proteintech, Cat# 25859-1-AP, RRID: AB_2716568), rabbit anti-zonula occludens-1 polyclonal antibody (ZO-1; 1:5000, Proteintech, Cat# 21773-1-AP, RRID: AB_10733242), rabbit anti-occludin polyclonal antibody (1:15,000, Proteintech, Cat# 27260-1-AP, RRID: AB_2880820), rabbit anti-AMPKα polyclonal antibody (1:1000, Cell Signaling Technology, Danvers, Massachusetts, USA, Cat# 2532, RRID: AB_330331), rabbit anti-phospho-AMPKα monoclonal antibody (1:1000, Cell Signaling Technology, Cat# 2535, RRID: AB_331250), and rabbit anti-PPARD polyclonal antibody (1:1000, Abcam, Cambridge, UK, Cat# ab23673, RRID: AB_2165902).

Techniques: Clinical Proteomics, Activation Assay, Expressing, Western Blot, Immunofluorescence, Staining, Control, Enzyme-linked Immunosorbent Assay, Binding Assay, Real-time Polymerase Chain Reaction

Journal: Neural Regeneration Research

Article Title: Chitosan alleviates symptoms of Parkinson’s disease by reducing acetate levels, which decreases inflammation and promotes repair of the intestinal barrier and blood–brain barrier

doi: 10.4103/NRR.NRR-D-23-01511

Figure Lengend Snippet: Chitosan may reduce acetate levels, thereby activating the PPARD-AMPK signaling pathway, which promotes repair of the intestinal barrier and reduces neuroinflammation in an MPTP-induced mouse model of PD. (A, B) Western blot analysis of p-AMPK, AMPK, and PPARD levels in mouse colon tissue ( n = 3/group). Treatment with acetate significantly increased p-AMPK and PPARD expression. (C) Treatment with a PPARD antagonist significantly decreased mouse body weight ( n = 6/group). (D) There were no significant differences in fall latency among the groups in the rotarod test, which was used to assess motor dysfunction ( n = 6/group). (E–G) PPARD antagonist treatment significantly decreased PPARD, TH, ZO-1, and occludin expression, as determined by western blot ( n = 3/group). (H) Immunofluorescence staining for ZO-1 (green, Alexa Fluor 488) and occludin (red, Alexa Fluor 594) in mouse colon tissue ( n = 3/group). The PPARD antagonist treatment group exhibited markedly reduced ZO-1 and occludin mRNA expression levels in colon tissue. Scale bars: 10 μm. (I) QPCR was used to measure the mRNA levels of IL-1β, IL-6, IL-8, IL-10, TNF-α, and iNOS in mouse colon tissue ( n = 3/group). Treatment with the PPARD antagonist increased IL-6 and TNF-α mRNA levels, while IL-8 and iNOS levels were reduced. (J) ELISA was used to detect IL-1β, IL-6, IL-10, and TNF-α expression levels in mouse plasma ( n = 5/group). IL-1β, IL-6, and TNF-α expression levels were significantly increased in the PPARD antagonist treatment group. (K) QPCR was used to measure mRNA levels of IL-1β, IL-6, IL-8, IL-10, TNF-α, and iNOS in the SN ( n = 3/group). Treatment with the PPARD antagonist significantly increased the mRNA levels of IL-1β, IL-6, and IL-8. (L) Treatment with the PPARD antagonist reduced p-AMPK, but not AMPK, expression ( n = 3/group). GAPDH was used as the internal reference. All data are presented as the mean ± SD. All experiments were repeated at least three times. * P < 0.05 (one-way analysis of variance followed by Tukey’s multiple comparisons test (A, B) or unpaired t -test (C–L)). AMPK: Adenosine 5′-monophosphate-activated protein kinase; DAPI: 4′,6-diamidino-2-phenylindole; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; IL-1β: interleukin-1 Beta; IL-6: interleukin-6; IL-8: interleukin-8; IL-10: interleukin-10; iNOS: inductible nitric oxide synthase; n.s.: not significant; NaA: sodium acetate; p-AMPK: phosphorylation adenosine 5′-monophosphate-activated protein kinase; PD: Parkinson’s disease; PPARD: peroxisome proliferator-activated receptor delta; QPCR: quantitative polymerase chain reaction; SN: substantia nigra; TH: tyrosine hydroxylase; TNF-α: tumor necrosis factor alpha; ZO-1: Zonula occludens-1.

Article Snippet: The primary antibodies used were as follows: rabbit anti-glyceraldehyde-3-phosphate dehydrogenase polyclonal antibody (GAPDH; 1:10,000, Proteintech, Wuhan, Hubei, China, Cat# 10494-1-AP, RRID: AB_2263076), rabbit anti-TH polyclonal antibody (1:5000, Proteintech, Cat# 25859-1-AP, RRID: AB_2716568), rabbit anti-zonula occludens-1 polyclonal antibody (ZO-1; 1:5000, Proteintech, Cat# 21773-1-AP, RRID: AB_10733242), rabbit anti-occludin polyclonal antibody (1:15,000, Proteintech, Cat# 27260-1-AP, RRID: AB_2880820), rabbit anti-AMPKα polyclonal antibody (1:1000, Cell Signaling Technology, Danvers, Massachusetts, USA, Cat# 2532, RRID: AB_330331), rabbit anti-phospho-AMPKα monoclonal antibody (1:1000, Cell Signaling Technology, Cat# 2535, RRID: AB_331250), and rabbit anti-PPARD polyclonal antibody (1:1000, Abcam, Cambridge, UK, Cat# ab23673, RRID: AB_2165902).

Techniques: Western Blot, Expressing, Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Phospho-proteomics, Real-time Polymerase Chain Reaction