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: Baicalin combined with metformin improved the blood glucose levels in metformin non-responsive mice by activating the AMPK/ACC/CPT1 pathway (A–F) The serum concentrations of (A) TC, (B) LDL-C, (C) HDL-C, (D) IL-1β, (E) IL-6, and (F) IL-10. (G) Hematoxylin-eosin (H&E) staining of the liver, pancreas, epididymal adipose tissues, and oil red O staining of the liver (magnification, 30×; scale bars, 100 μm). (H) Western blot analysis of pAMPK (Thr172), ACC, and CPT1 proteins in the livers of mice after administration. Data are expressed as the mean ± SD (A‒F, n = 6–8; H, n = 3); ∗ 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 (A, B, C, E, F, and H) and Kruskal-Wallis test (D). NCD, normal chow diet; NR, non-response; Met, metformin; BA, baicalin; TC, total cholesterol; LDL-C, low-density lipoprotein cholesterol; HDL-C, high-density lipoprotein cholesterol; IL-1β, interleukin-1β; IL-6, interleukin-6; IL-10, interleukin-10; AMPK, AMP-activated protein kinase; ACC, acetyl-CoA carboxylase; CPT1, carnitine palmitoyl transferase 1.

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: Staining, Western Blot

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 treatment reversed the metformin NR phenotype in NR mice (A) Experimental protocol for administration of R. hominis in mice. (B) Oral glucose tolerance test (OGTT) curve and its area under the curve (AUC). (C) Insulin tolerance test (ITT) curve and its AUC. (D) Homeostasis model assessment of insulin resistance (HOMA-IR) after drug R. hominis . (E–J) The serum concentrations of (E) TC, (F) LDL-C, (G) HDL-C, (H) IL-1β, (I) IL-6, and (J) IL-10. (K) Hematoxylin-eosin (H&E) staining of the liver, pancreas, epididymal adipose tissues, and oil red O staining of the liver (magnification, 30×; scale bars, 100 μm). (L) Western blot analysis of AMPK/ACC/CPT1 proteins in the livers of mice after 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; ABX, antibiotic mixed; FMT, fecal microbial transplantation; NCD, normal chow diet; R.h, Roseburia hominis ; TC, total cholesterol; LDL-C, low-density lipoprotein cholesterol; HDL-C, high-density lipoprotein cholesterol; IL-1β, interleukin-1β; IL-6, interleukin-6; IL-10, interleukin-10; AMPK, AMP-activated protein kinase; ACC, acetyl-CoA carboxylase; CPT1, carnitine palmitoyl transferase 1.

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: Staining, Western Blot, Transplantation Assay

Journal: Chinese medicine

Article Title: Salidroside protects against myocardial infarction via activating MIF-mediated mitochondrial quality control.

doi: 10.1186/s13020-025-01076-3

Figure Lengend Snippet: Fig. 2 Salidroside activated MIF signaling and decreased mitochondrial damage in myocardial infarction model mice. A Salidroside was administered at various concentrations following MI surgery, and heart tissue samples were collected. The mitochondrial ultrastructure was observed via transmission electron microscopy, and damaged mitochondria were quantified. The yellow or blue regions indicate damaged mitochondria or intact mitochondria, respectively, scale bar = 2 or 1 μm. B The ratio of mtDNA to nuclear DNA was determined via quantitative real-time PCR. C ATP content was measured. D Western blot analysis of the expression of MIF. E Representative images of MIF immunofluorescence staining and data analysis of heart sections. The cell nuclei were stained blue with DAPI, and MIF was stained red, scale bar = 100 μm. F Western blot analysis of the expression of p-AMPK, LC3I, LC3II, PINK1, BNIP3, OPA1, and MFN1 were performed. Statistical analysis was performed via one-way ANOVA followed by Bonferroni’s multiple comparisons test. n = 3. *P < 0.05 vs. the MI-Ctrl group; **P < 0.01 vs. the MI-Ctrl group; #P < 0.05 vs. the MI-Sal-L group; ##P < 0.01 vs. the MI-Sal-L group

Article Snippet: After sodium dodecyl sulfate‒polyacrylamide gel electrophoresis (SDS‒PAGE), electrophoretic transfer, and nonspecific binding blocking, the transferred proteins were incubated overnight with primary antibodies against GAPDH (1:5000, 10494-1-AP, Proteintech), Bcl-2 (1:2000, ab182858, Abcam), Bax (1:2000, ab32503, Abcam), cleaved caspase-3 (1:1000, #9661, CST), MIF (1:1000, ab187064, Abcam), AMPK (1:1500, 66536-1-Ig, Proteintech), p-AMPK (1:1000, ab23875, Abcam), microtubule-associated protein 1 light chain 3 II (LC3II, 1:1000, 14600-1-AP, Proteintech), PTEN-induced putative kinase 1 (PINK1, 1:1000, 23274-1-AP, Proteintech), BCL2interacting protein 3 (BNIP3, 1:1000, ab109362, Abcam), Optic Atrophy 1 (OPA1, 1:2000, 27733-1-AP, Proteintech), and Mitofusin 1 (MFN1, 1:1000, 2398-1-AP, Proteintech), followed by incubation with the corresponding Band intensity was analyzed using a gel documentation system (Bio-Rad, Hercules, CA, USA) and normalized to that of GAPDH.

Techniques: Transmission Assay, Electron Microscopy, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Immunofluorescence, Staining

Journal: Chinese medicine

Article Title: Salidroside protects against myocardial infarction via activating MIF-mediated mitochondrial quality control.

doi: 10.1186/s13020-025-01076-3

Figure Lengend Snippet: Fig. 5 Effects of MIF on oxygen‒glucose deprivation (OGD)-induced apoptosis and mitochondrial dysfunction in cardiomyocytes. A Cardiomyocytes were subjected to oxygen‒glucose deprivation (OGD) or control conditions and transfected with siMif or incubated with recombinant MIF (rMIF) for 24 h. Then, western blot analysis of the expression of MIF was performed. B Western blot analysis of the expression of Bax, Bcl-2, and caspase-3 was performed. C Cardiomyocyte apoptosis was detected and quantified by flow cytometry. D Mitochondria were quantified by mitochondrial labeling, and the data were analyzed. Scale bar = 100 μm. E ATP content was measured. F Western blot analysis of p-AMPK, LC3-I, LC3-II, PINK1, BNIP3, OPA1, and MFN1 was performed. The data are presented as the means ± SDs. Statistical analysis was performed via one-way ANOVA followed by Bonferroni’s multiple comparisons test. n = 3. *P < 0.05 vs. the Ctrl group, **P < 0.01 vs. the Ctrl group, #P < 0.05 vs. the OGD group, ##P < 0.01 vs. the OGD group

Article Snippet: After sodium dodecyl sulfate‒polyacrylamide gel electrophoresis (SDS‒PAGE), electrophoretic transfer, and nonspecific binding blocking, the transferred proteins were incubated overnight with primary antibodies against GAPDH (1:5000, 10494-1-AP, Proteintech), Bcl-2 (1:2000, ab182858, Abcam), Bax (1:2000, ab32503, Abcam), cleaved caspase-3 (1:1000, #9661, CST), MIF (1:1000, ab187064, Abcam), AMPK (1:1500, 66536-1-Ig, Proteintech), p-AMPK (1:1000, ab23875, Abcam), microtubule-associated protein 1 light chain 3 II (LC3II, 1:1000, 14600-1-AP, Proteintech), PTEN-induced putative kinase 1 (PINK1, 1:1000, 23274-1-AP, Proteintech), BCL2interacting protein 3 (BNIP3, 1:1000, ab109362, Abcam), Optic Atrophy 1 (OPA1, 1:2000, 27733-1-AP, Proteintech), and Mitofusin 1 (MFN1, 1:1000, 2398-1-AP, Proteintech), followed by incubation with the corresponding Band intensity was analyzed using a gel documentation system (Bio-Rad, Hercules, CA, USA) and normalized to that of GAPDH.

Techniques: Control, Transfection, Incubation, Recombinant, Western Blot, Expressing, Flow Cytometry, Labeling

Journal: Chinese medicine

Article Title: Salidroside protects against myocardial infarction via activating MIF-mediated mitochondrial quality control.

doi: 10.1186/s13020-025-01076-3

Figure Lengend Snippet: Fig. 6 Salidroside alleviates mitochondrial dysfunction and apoptosis in oxygen‒glucose deprivation (OGD)-induced cardiomyocytes via MIF signaling. A Cardiomyocytes were transfected with siMif or treated with recombinant MIF and then treated with salidroside or PBS with or without oxygen‒glucose deprivation (OGD) for 24 h. Proteins were subsequently harvested for western blotting to determine the level of MIF. B Western blotting was performed to determine the expression levels of Bax, Bcl-2, and cleaved caspase 3. C Cardiomyocyte apoptosis was detected and quantified by flow cytometry. D ATP content was measured. E Mitochondria were quantified by mitochondrial labeling. Scale bar = 100 μm. F, G Western blot analysis of the levels of p-AMPK, LC3I, LC3II, PINK1, BNIP3, OPA1, and MFN1 was performed. The data are presented as the means ± SDs. Statistical analysis was performed via one-way ANOVA followed by Bonferroni’s multiple comparisons test. n = 3. *P < 0.05 vs. the Ctrl group, **P < 0.01 vs. the Ctrl group, #P < 0.05 vs. the OGD group, ##P < 0.01 vs. the OGD group, @P < 0.05 vs. the Sal group, @@P < 0.01 vs. the Sal group

Article Snippet: After sodium dodecyl sulfate‒polyacrylamide gel electrophoresis (SDS‒PAGE), electrophoretic transfer, and nonspecific binding blocking, the transferred proteins were incubated overnight with primary antibodies against GAPDH (1:5000, 10494-1-AP, Proteintech), Bcl-2 (1:2000, ab182858, Abcam), Bax (1:2000, ab32503, Abcam), cleaved caspase-3 (1:1000, #9661, CST), MIF (1:1000, ab187064, Abcam), AMPK (1:1500, 66536-1-Ig, Proteintech), p-AMPK (1:1000, ab23875, Abcam), microtubule-associated protein 1 light chain 3 II (LC3II, 1:1000, 14600-1-AP, Proteintech), PTEN-induced putative kinase 1 (PINK1, 1:1000, 23274-1-AP, Proteintech), BCL2interacting protein 3 (BNIP3, 1:1000, ab109362, Abcam), Optic Atrophy 1 (OPA1, 1:2000, 27733-1-AP, Proteintech), and Mitofusin 1 (MFN1, 1:1000, 2398-1-AP, Proteintech), followed by incubation with the corresponding Band intensity was analyzed using a gel documentation system (Bio-Rad, Hercules, CA, USA) and normalized to that of GAPDH.

Techniques: Transfection, Recombinant, Western Blot, Expressing, Flow Cytometry, Labeling