Journal: Acta Pharmaceutica Sinica. B

Article Title: Gentiopicroside targets PAQR3 to activate the PI3K/AKT signaling pathway and ameliorate disordered glucose and lipid metabolism

doi: 10.1016/j.apsb.2021.12.023

Figure Lengend Snippet: GPS treatment promoted the activation of the PI3K/AKT axis to improve glucose and lipid metabolism in PA-treated HepG2 cells. (A) Glucose uptake assessment in GPS-treated HepG2 cells under palmitate acids conditions ( n = 3). ∗∗∗ P < 0.001 vs. control group, ## P < 0.01 vs. PA-induced group. (B) TG and TC contents in HepG2 cells ( n = 3). ∗∗ P < 0.01 vs. control group, ## P < 0.01 vs. PA-treated group. (C) Lipid droplets deposition was assessed by oil red staining and glycogen synthesis was assessed by PAS staining. (D) GCK, G6Pase, LDLR and SREBP-1c protein levels in palmitate acid-induced HepG2 cells ( n = 3). ∗∗∗ P < 0.001, ∗∗ P < 0.01 vs. control group; ### P < 0.001, ## P < 0.01, # P < 0.05 vs. PA-induced group. (E) Effect of GPS on the phosphorylation level of PI3K, AKT and GSK3 β ( n = 3). ∗∗ P < 0.01 vs. control group; ## P < 0.01 vs. insulin-induced group; ^^ P < 0.01, ^ P < 0.05 vs. PA + insulin-induced group. (F) PI3K activity in GPS co-treated HepG2 cells and (G) PIP3 levels in GPS co-treated HepG2 cells ( n = 3). ∗∗∗ P < 0.001 vs. control group; ### P < 0.001, ## P < 0.01 vs. insulin-induced group; ^^ P < 0.01, ^ P < 0.05 vs. PA + insulin-induced group, ns: no significance. (H) The subcellular distribution of FOXO1 and SREBP-1c was determined by immunofluorescent. Green fluorescence indicates FOXO1, red fluorescence indicates SREBP-1c and blue fluorescence indicates nucleus, scale bar = 20 μm. (I) Phosphorylation of FOXO1 and nuclear distribution of FOXO1 and SREBP-1c were measured by Western blot ( n = 3). ∗∗ P < 0.01, ∗ P < 0.05 vs. control group; ### P < 0.001, ## P < 0.01 vs. insulin-induced group; ^ P < 0.01, ^ P < 0.05 vs. PA + insulin-induced group. Protein level was quantified and normalized to α -tubulin in the control group cells. The data are presented as mean ± standard deviation (SD), all experiments were performed at least three times with similar results. ns, no significance.

Article Snippet: PAS staining was carried out with a commercial kit (Cat.: G1281, Solarbio, Beijing, China) according to the manufacturer's protocol, and the images were captured by EVOS FL Auto system (Life TechnologiesTM, Grand Island, NY, USA).

Techniques: Activation Assay, Control, Staining, Phospho-proteomics, Activity Assay, Fluorescence, Western Blot, Standard Deviation

Journal: Gut and Liver

Article Title: The Use of Foxa2-Overexpressing Adipose Tissue-Derived Stem Cells in a Scaffold System Attenuates Acute Liver Injury

doi: 10.5009/gnl18235

Figure Lengend Snippet: Hepatic differentiation ability in two-dimensional (2D) culture. (A) Gene expression of hepatic differentiation markers (AFP, ALB, and CK18) in transfected ADSCs after 2 weeks measured by qRT-PCR. (B) Amount of cytoplasmic glycogen after the overexpression of various gene in 2D culture measured by periodic acid-Schiff (PAS) staining (PAS staining: blue, nuclei; purple, glycogen in cytoplasm; magnification: ×400). The arrows indicate cytosol. AFP, alpha fetoprotein, ALB, albumin; CK18, cytokeratin 18; ADSCs, adipose tissue-derived stem cells; qRT-PCR, real-time quantitative polymerase chain reaction; Foxa2, forkhead box protein. *p<0.05.

Article Snippet: For liver function evaluation, periodic acid-Schiff staining (PAS Staining Kit, 1.01646.0001; Merck Millipore) was performed to analyze glycogen storage capacity.

Techniques: Gene Expression, Transfection, Quantitative RT-PCR, Over Expression, Staining, Derivative Assay, Real-time Polymerase Chain Reaction

Journal: Gut and Liver

Article Title: The Use of Foxa2-Overexpressing Adipose Tissue-Derived Stem Cells in a Scaffold System Attenuates Acute Liver Injury

doi: 10.5009/gnl18235

Figure Lengend Snippet: Comparison of hepatic differentiation ability in three-dimensional (3D) culture using a scaffold. (A) qRT-PCR expression of hepatic differentiation markers (AFP, CK18, CD26, and CX32) in Foxa2-transfected ADSCs after 2 weeks in 3D culture. (B) H&E staining, periodic acid-Schiff (PAS) and cytochrome P450 immunofluorescence in Foxa2-overexpressing ADSCs in 3D culture (a and b: H&E staining, blue - nuclei, pink - cytosol, ×400; c and d: PAS staining, blue - nuclei, purple - glycogen in cytoplasm, ×1,000; e and f: blue - nuclei, green - cytochrome 450 in cytosol, ×1,000). (C) Quantification of cytochrome P450 intensity and glucose expression. qRT-PCR, real-time quantitative polymerase chain reaction; AFP, alpha fetoprotein; CK18, cytokeratin 18; CD26, dipeptidyl peptidase 4; CX32, connexin 32; ADSCs, adipose tissue-derived stem cells; Foxa2, forkhead box protein. *p<0.05.

Article Snippet: For liver function evaluation, periodic acid-Schiff staining (PAS Staining Kit, 1.01646.0001; Merck Millipore) was performed to analyze glycogen storage capacity.

Techniques: Comparison, Quantitative RT-PCR, Expressing, Transfection, Staining, Immunofluorescence, Real-time Polymerase Chain Reaction, Derivative Assay

Journal: Gut and Liver

Article Title: The Use of Foxa2-Overexpressing Adipose Tissue-Derived Stem Cells in a Scaffold System Attenuates Acute Liver Injury

doi: 10.5009/gnl18235

Figure Lengend Snippet: Comparison of hepatocyte differentiation in a scaffold during in vivo culture. (A) In vivo culture design of scaffold preparation and implantation into nude mice. (B) Scanning electron microscopy of ADSCs in the PLGA scaffold. (C) qRT-PCR expression of hepatic differentiation markers (AFP, CK18, CD26 and CX32) in Foxa2-transfected ADSCs after 2 weeks in nude mice. (D) H&E staining, periodic acid-Schiff (PAS) staining and immunofluorescent staining for cytochrome P450 activity after Foxa2 gene overexpression in in vivo culture (a and b: H&E staining, blue - nuclei, pink - cytosol, ×400; c and d: PAS staining, blue - nuclei, purple - glycogen in cytoplasm, ×1,000; e and f: blue - nuclei, green - cytochrome 450 in cytosol, ×1,000). (E) Quantification of cytochrome P450 intensity and glucose expression. ADSCs, adipose tissue-derived stem cells; PLGA, poly(lactic-co-glycolic acid); qRT-PCR, real-time quantitative polymerase chain reaction; AFP, alpha fetoprotein; CK18, cytokeratin 18; CD26, dipeptidyl peptidase 4; CX32, connexin 32; Foxa2, forkhead box protein. *p<0.05.

Article Snippet: For liver function evaluation, periodic acid-Schiff staining (PAS Staining Kit, 1.01646.0001; Merck Millipore) was performed to analyze glycogen storage capacity.

Techniques: Comparison, In Vivo, Electron Microscopy, Quantitative RT-PCR, Expressing, Transfection, Staining, Activity Assay, Over Expression, Derivative Assay, Real-time Polymerase Chain Reaction

Journal: Gut and Liver

Article Title: The Use of Foxa2-Overexpressing Adipose Tissue-Derived Stem Cells in a Scaffold System Attenuates Acute Liver Injury

doi: 10.5009/gnl18235

Figure Lengend Snippet: Stem cell homing. (A) Mouse and human β2-microglobulin mRNA expression was compared between the TAA-only group and the ADSCs scaffold group. (B) Immunofluorescence stain using human anti-mitochondria antibody was performed in mouse liver (×200) and implanted scaffold (×630) (immunofluorescence: blue - nuclei, green - human mitochondria). H&E staining (blue - nuclei, pink - cytosol, ×400) and periodic acid-Schiff (PAS) staining (blue - nuclei, purple - glycogen in cytoplasm, ×630) of implanted scaffold. TAA, thioacetamide; ADSCs, adipose tissue-derived stem cells; Foxa2, forkhead box protein.

Article Snippet: For liver function evaluation, periodic acid-Schiff staining (PAS Staining Kit, 1.01646.0001; Merck Millipore) was performed to analyze glycogen storage capacity.

Techniques: Expressing, Immunofluorescence, Staining, Derivative Assay