Journal: Journal of Pharmaceutical Analysis

Article Title: Chemical characterization of aconite polysaccharide and its ameliorative effect on ulcerative colitis by regulating gut microbiota and THDCA-driven TGR5/cAMP/PKA/NLRP3 pathway

doi: 10.1016/j.jpha.2026.101642

Figure Lengend Snippet: Impact of taurohyodeoxycholic acid (THDCA) and aconite polysaccharide A-1 (APA-1) on Takeda G protein-coupled receptor 5 (TGR5)/cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) signaling and nucleotide-binding oligomerization domain (NOD), leucine-rich repeat and pyrin domain containing 3 (NLRP3) inflammasome in ulcerative colitis (UC) mice. (A) Representative Western blotting (WB) images of TGR5, cAMP-dependent protein kinase catalytic subunit, apoptosis-associated speck-like protein containing a CARD (ASC), NLRP3, and interleukin-1β (IL-1β) P17 of Normal, dextran sulfate sodium (DSS), and THDCA group mice. (B) Relative protein expression levels of colonic TGR5, cAMP-dependent protein kinase catalytic subunit, ASC, NLRP3, and IL-1β P17 of Normal, DSS, and THDCA group mice. (C) Representative WB images of TGR5, cAMP-dependent protein kinase catalytic subunit, NLRP3, ASC, and IL-1β P17 of Control, Model, APA-1-low dose (APA-1-L), and APA-1-high dose (APA-1-H) group mice. (D) Relative protein expression levels of colonic TGR5, cAMP-dependent protein kinase catalytic subunit, NLRP3, ASC, and IL-1β P17 of Control, Model, APA-1-L, and APA-1-H group mice. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001. GAPDH: glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: The membranes were then incubated overnight with the following primary antibodies: anti-claudin1 (1:5000; Proteintech, Wuhan, China), anti-occludin (1:15,000; Proteintech), anti-interleukin-6 (IL-6) (1:5000; Servicebio Technology Co., Ltd.), anti-tumor necrosis factor-alpha (TNF-α) (1:2000; Abmart, Shanghai, China), anti-Takeda G protein-coupled receptor 5 (TGR5) (1:2000; Abclonal, Wuhan, China), anti-cyclic adenosine monophosphate (cAMP)-dependent protein kinase catalytic subunit (1:20,000; Abcam, Cambridge, UK), anti-nucleotide-binding domain, leucine-rich-repeat-containing family, pyrin domain-containing 3 (NLRP3) (1:5000; Abclonal), anti-apoptosis-associated speck-like protein containing a CARD (ASC) (1:10,000; Abclonal), anti-IL-1β P17 (1:2000; Abcam), anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:5000; Servicebio Technology Co., Ltd.), and anti-tubulin (1:10,000; Abclonal).

Techniques: Binding Assay, Western Blot, Expressing, Control