Journal: iScience
Article Title: Roseburia hominis enriched by baicalin reverses the non-response to metformin via upregulating linolenic acid metabolism
doi: 10.1016/j.isci.2025.113892
Figure Lengend Snippet: R. hominis intervention promoted LA metabolism in mice, and its effect on reversing the NR in mice depended on PLA2 (A) The PCA score plot in negative and positive ion modes. In the negative ion mode, the prediction rate (Q2) of the PCA-X model is 0.599, and the interpretation rate (R2X) is 0.771. The prediction rate (Q2) of the PCA-X model in positive ion mode is 0.487, and the interpretation rate (R2X) is 0.712. (B) Enrichment analysis of differential metabolites. (C–E) The serum abundance of (C) PC (18:3/16:0), (D) LA, and (E) SDA. (F) Schematic diagram of LA metabolism. The blue background represents metabolites, while the green background represents enzymes. The red font represents the substances detected in the study. (G) Western blot analysis of PLA2G and FADS2 proteins in the livers of mice after R. hominis administration. (H) The activity of PLA2 after darapladib intervention. (I) Oral glucose tolerance test (OGTT) curve and its area under the curve (AUC). (J) Insulin tolerance test (ITT) curve and its AUC. (K) Homeostatic model assessment for insulin resistance (HOMA-IR) after drug administration. Data are expressed as the mean ± SD ( n = 7–8); ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, as determined by one-way ANOVA with Holm-Sidak’s post hoc test. ANOVA, analysis of variance; PC, phosphatidylcholine; NR, non-response; NCD, normal chow diet; R.h, Roseburia hominis ; PLA2, phospholipase A2; FADS2, fatty acid desaturase 2; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; PCA, principal-component analysis.
Article Snippet: These membranes were then incubated at 4°C overnight with primary antibodies against AMP-activated protein kinase (AMPK) (Proteintech, Cat# 10929-2-AP, RRID: AB_2169568 , 1:4,000), insulin receptor substrate 1 (IRS1) (Proteintech, Cat# 17509-1-AP, RRID: AB_10596914 , 1:1,000), phospho-AMPKα (Thr172) (40H9) (Cell Signaling Technology, Cat# 2535, RRID: AB_331250 , 1:1,000), phospho-IRS-1 (Ser636/639) (Cell Signaling Technology, Cat# 2388, RRID: AB_330339 , 1:1,000), phosphoenolpyruvate carboxykinase (PCK) (Proteintech, Cat# 16754-1-AP, RRID: AB_2160031 , 1:30,000), glucose-6-phosphatase (G6PC) (Proteintech, Cat# 66860-1-Ig, RRID: AB_2882199 , 1:5,000), mammalian target of rapamycin (mTOR) (Proteintech, Cat# 66888-1-Ig, RRID: AB_2882219 , 1:30,000), fatty acid desaturase 2 (FADS2) (Proteintech, Cat# 28034-1-AP, RRID: AB_2918142 , 1:4,000), phospholipase A2 group (PLA2G) (Proteintech, Cat# 18088-1-AP, RRID: AB_10859777 , 1:1,000), acetyl-CoA Carboxylase 1 (ACC1) (Proteintech, Cat# 21923-1-AP, RRID: AB_11042445 , 1:4,000), carnitine palmitoyl transferase 1 (CPT1) (Proteintech, Cat# 15184-1-AP, RRID: AB_2084676 , 1:50,000), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Proteintech, Cat# 10494-1-AP, RRID: AB_2263076 , 1:20,000), β-tubulin (Proteintech, Cat# 10094-1-AP, RRID: AB_2210695 , 1:10,000).
Techniques: Western Blot, Activity Assay