Journal: International Journal of Molecular Sciences

Article Title: Protective Effect of Paeoniae Radix Alba Carbonisata on Hepatic Amyloidosis by Regulating Calcium Homeostasis

doi: 10.3390/ijms27062582

Figure Lengend Snippet: Transcriptomic results of PRAC-E ameliorating HA. ( A ) Venn diagram of the DEGs from PRAC-E vs. LYSO-6 and LYSO-6 vs. Normal. ( B ) LYSO-6 vs. Normal DEGs volcano plot. ( C ) PRAC-E vs. LYSO-6 DEGs volcano plot. ( D ) Functional enrichment analysis of downregulated DEGs in LYSO-6 vs. Normal. ( E ) Function enrichment analysis of upregulated DEGs in PRAC-E vs. LYSO-6. ( F ) Pathway enrichment analysis of the downregulated DEGs in LYSO-6 vs. Normal. ( G ) Pathway enrichment analysis of upregulated DEGs in PRAC-E vs. LYSO-6. ( H ) Ca 2+ -ATPase activity in liver tissue ( n = 6). Data are presented as the mean ± standard deviation (SD) ( n = 6). ** p < 0.01, *** p < 0.001 vs. LYSO-6.

Article Snippet: ALT, AST, ALP, and ultramicro Ca 2+ -ATPase detection kits were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).

Techniques: Functional Assay, Activity Assay, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Protective Effect of Paeoniae Radix Alba Carbonisata on Hepatic Amyloidosis by Regulating Calcium Homeostasis

doi: 10.3390/ijms27062582

Figure Lengend Snippet: PRAC-E modulates calcium homeostasis via the cGMP/PKG/ATP2A1 signaling axis, thereby alleviating inflammation and amyloid deposition. ( A , B ) The dose-dependent ( A ) and time-dependent ( B ) effects of LYSO-6 on NCTC1469 cells. ( C ) Screening of the effective concentration of PRAC-E on LYSO-6-stimulated cells. ( D – F ) The levels of IL-6, TNF-α, and IL-8 in the cell culture supernatant. ( G – I ) The levels of SAA, SAP ( G ), and APO-E ( H ) in NCTC1469 cells. ( J ) CR staining of NCTC1469 cells. ( K ) The dose-dependent effects of BAPTA-AM on NCTC1469 cells. ( L , M ) Representative Fluo-4 AM fluorescence staining and semi-quantification of NCTC1469 cells. ( N ) Ca 2+ -ATPase activity in NCTC1469 cells. ( O ) Intracellular cGMP levels in NCTC1469 cells were measured by ELISA. ( P – R ) Western blotting and semi-quantitative analysis of PDE5A, PKG, and ATP2A1 in NCTC1469 cells. Data are presented as the mean ± SD ( n = 6). # p < 0.05, ## p < 0.01, ### p < 0.001 vs. solvent control group; * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. LYSO-6; ns, no significance.

Article Snippet: ALT, AST, ALP, and ultramicro Ca 2+ -ATPase detection kits were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China).

Techniques: Concentration Assay, Cell Culture, Staining, Fluorescence, Activity Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Solvent, Control

Journal: Foods

Article Title: Evolution of Physicochemical Properties and Volatile Organic Compound Profiles in Pre-Cooked Braised Chicken During Storage

doi: 10.3390/foods15010091

Figure Lengend Snippet: Changes in pH, malondialdehyde (MDA) content, and Ca 2+ -ATPase activity of pre-cooked braised chicken during storage. ( a ) pH value, ( b ) MDA content, ( c ) Ca 2+ -ATPase activity. Note: Different lowercase letters above bars indicate statistically significant differences ( p < 0.05).

Article Snippet: The diluted supernatant was combined with reagents from a commercial Ca 2+ -ATPase assay kit(Jiancheng Bioengineering Institute Co., Ltd., Nanjing, China), following the manufacturer’s instructions.

Techniques: Activity Assay

Journal: Journal of Pharmaceutical Analysis

Article Title: Unlocking the potential of atractylenolide II: Mitigating non-alcoholic fatty liver disease through farnesoid X receptor-endoplasmic reticulum stress interplay

doi: 10.1016/j.jpha.2025.101318

Figure Lengend Snippet: Atractylenolide II (AT-II) activates farnesoid X receptor (FXR)-sarco/endoplasmic reticulum Ca 2+ ATPase 2 (SERCA2) axis in hepatocytes. (A–D) HepG2 or AML-12 cells were treated with AT-II or GW4064 (5 μM) for 24 h before harvesting, with dimethyl sulfoxide (DMSO) treatment as negative control. n = 3. (A) Relative messenger RNA (mRNA) expression of SERCA2 in HepG2 cells. (B, C) Protein levels of SERCA2 in HepG2 cells were analyzed by Western blotting (B), and quantified (C). (D) Immunofluorescence detection of SERCA2 (red) in HepG2 or AML-12 cells. (E–G) HepG2 cells receiving a 48-h FXR siRNA transfection were subjected to the detection of relative mRNA (E), and protein (F, G) expression of SERCA2. n = 3. (H) HepG2 cells were treated with AT-II or GW4064 (5 μM) for 24 h before harvesting. Ca 2+ ATPase activity was measured. n = 5. (I) Predicted FXR binding motifs in the −3000 SERCA2 promoter region. (J) An illustration of constructed pGL3-SERCA2 promoter plasmids and its mutants for luciferase reporter assays. (K) HepG2 cells were transfected with pGL3-SERCA2 promoter and its mutants, with or without hFXR overexpression. The activities of corresponding promoters were assessed via luciferase reporter assays. n = 5. (L) HepG2 cells were overexpressed with or without hFXR. Chromatin immunoprecipitation (ChIP) analysis of FXR binding to the SERCA2 promoter region in HepG2 cells were analyzed. n = 5. (M, N) HepG2 cells were over expressed with pGL3-SERCA2 and hFXR plasmids, and then treated with 0.1 % DMSO, 10 μM of chenodeoxycholic acid (CDCA), 10 μM of obeticholic acid (OCA), GW4064, and AT-II. pGL3-SERCA2 reporter analysis were conducted using resuling cells. n = 5. Data were indicated as the means ± standard error of the mean (SEM). ∗ P < 0.05, vs. control group or si-control group, NS: no significance. RLU: relative light unit; IgG: immunoglobulin G.

Article Snippet: The intracellular Ca 2+ ATPase activity was measured using an Ultramicro Ca 2+ ATPase Diagnostic Kit (Cat# A070-4-2, Nanjing Jiancheng, Nanjing, China).

Techniques: Negative Control, Expressing, Western Blot, Immunofluorescence, Transfection, Activity Assay, Binding Assay, Construct, Luciferase, Over Expression, Chromatin Immunoprecipitation, Control

Journal: Journal of Pharmaceutical Analysis

Article Title: Unlocking the potential of atractylenolide II: Mitigating non-alcoholic fatty liver disease through farnesoid X receptor-endoplasmic reticulum stress interplay

doi: 10.1016/j.jpha.2025.101318

Figure Lengend Snippet: Activation of the farnesoid X receptor (FXR)-sarco/endoplasmic reticulum Ca 2+ ATPase 2 (SERCA2) axis in hepatic cells by atractylenolide II (AT-II) in diet-induced obese (DIO) mice. A high-fat diets (HFD) with or without AT-II (100 mg/100 g diet) was administered to obese male C57BL/6 mice for six weeks. (A) Hepatic total bile acid (TBA) levels. n = 7. (B) Hepatic expression of FXR signaling genes were measured by quantitative real-time polymerase chain reaction (qRT-PCR). n = 5. (C, D) Nuclear translocation of liver FXR was analyzed by Western blotting (C), and quantified (D). n = 6. Histone H3 (H3) and actin beta (ACTB) were used as internal references for nuclear and cytoplasmic protein expression, respectively. (E, F) Hepatic small heterodimer partner (SHP) protein levels were imaged by Western blotting (E), and quantified (F). n = 6. (G) Immunofluorescence detection of SERCA2 in the liver. (H–J) Hepatic messenger RNA (mRNA) levels of Serca2 (H), C/EBP homologous protein ( Chop ) (I), and glucose-regulated protein 78 ( Bip ) were detected by quantitative real-time polymerase chain reaction (qRT-PCR). n = 5. (K, L) Protein expression of heptic SERCA2 and endoplasmic reticulum (ER) stress signaling were detected by Western blotting (K), and quantified (L). n = 6. (M) Hepatic lipid metabolism gene expression. n = 5. (N) Hepatic total malondialdehyde (MDA) levels. n = 7. (O) Relative mRNA expression of oxidative stress-related genes in the liver. n = 5. All data were indicated as the means ± standard error of the mean (SEM). ∗ P < 0.05, HFD vs. Chow; # P < 0.05, HFD vs. HFD + AT-II. ABCC2: ATP-binding cassette sub-family C member 2; ABCB11: ATP-binding cassette sub-family B member 11; ABCB4: ATP-binding cassette sub-family B member 4; CYP7A1: cytochrome p450 family 7 subfamily a member 1; cle-CASP3: cleaved Caspase-3; CPT1α: carnitine palmitoyltransferase 1 α; CEBP: CCAAT/enhancer binding protein; SCD1: stearoyl-CoA desaturase 1; GPX: glutathione peroxidase; GCLC: glutamate-cysteine ligase catalytic subunit; SOD2: superoxide dismutase 2.

Article Snippet: The intracellular Ca 2+ ATPase activity was measured using an Ultramicro Ca 2+ ATPase Diagnostic Kit (Cat# A070-4-2, Nanjing Jiancheng, Nanjing, China).

Techniques: Activation Assay, Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Translocation Assay, Western Blot, Immunofluorescence, Gene Expression, Binding Assay

Journal: Journal of Pharmaceutical Analysis

Article Title: Unlocking the potential of atractylenolide II: Mitigating non-alcoholic fatty liver disease through farnesoid X receptor-endoplasmic reticulum stress interplay

doi: 10.1016/j.jpha.2025.101318

Figure Lengend Snippet: Adeno-associated virus 8 (AAV8)-mediated sarco/endoplasmic reticulum Ca 2+ ATPase 2 ( Serca2 ) silencing in the liver diminishes the efficacy of atractylenolide II (AT-II) against insulin resistance and fatty liver in diet-induced obese (DIO) mice. (A) Schematic of the animal model design. (B–E) Obese male C57BL/6 mice received tail vein injections of AAV8-shNC or AAV8-sh Serca2 and were then fed a high-fat diets (HFD) with or without AT-II (100 mg/100 g diet) for six weeks. n = 6. (B) Body weight gain. (C) Fasting blood glucose. (D) Intraperitoneal insulin tolerance test (IPITT). (E) Enzyme-linked immunosorbent assay (ELISA) measurement of serum insulin levels. (F–K) The liver tissues from mice with AAV8 interference were subjected to hematoxylin and eosin (H&E) and Oil-Red O (ORO) staining (F), scoring of steatosis (G), lobular inflammation (H), hepatocyte ballooning (I), and non-alcoholic fatty liver disease activity score (NAS) (J), and ORO quantification (K). n = 6. (L) Total liver triglyceride (TG) content. n = 6. (M, N) Apoptosis in the liver tissues was analyzed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) (M), and the TUNEL positive cells were quantified (N). n = 6. (O, P) The serum from mice with AAV8 interference were subjected to analysis of alanine aminotransferase (ALT) (O), and aspartate aminotransferase (AST) (P). n = 6. All data were shown as the means ± standard error of the mean (SEM). ∗ P < 0.05, AAV8-shNC HFD vs. AAV8-shNC HFD + AT-II; # P < 0.05, AAV8-sh Serca2 HFD vs. AAV8-sh Serca2 HFD + AT-II; NS: no significance. ND: not detected; NC: negative control; FBG: fasting blood glucose.

Article Snippet: The intracellular Ca 2+ ATPase activity was measured using an Ultramicro Ca 2+ ATPase Diagnostic Kit (Cat# A070-4-2, Nanjing Jiancheng, Nanjing, China).

Techniques: Virus, Animal Model, Enzyme-linked Immunosorbent Assay, Staining, Activity Assay, TUNEL Assay, Negative Control

Journal: Journal of Pharmaceutical Analysis

Article Title: Unlocking the potential of atractylenolide II: Mitigating non-alcoholic fatty liver disease through farnesoid X receptor-endoplasmic reticulum stress interplay

doi: 10.1016/j.jpha.2025.101318

Figure Lengend Snippet: Sarco/endoplasmic reticulum Ca 2+ ATPase 2 (SERCA2)-mediated eukaryotic translation initiation factor 2 subunit α (eIF2α) dephosphorylation in hepatocytes contributes to the anti-non-alcoholic fatty liver disease (NAFLD) effects of atractylenolide II (AT-II). (A–C) Obese male C57BL/6 mice injected with adeno-associated virus 8 (AAV8)-shNC and AAV8-sh Serca2 via the tail vein injection were fed a high-fat diets (HFD) with or without AT-II (100 mg/100 g diet) for 6 weeks. n = 6. The liver tissues were collected for determination of protein expression by Western blotting (A), and quantified (B), and periodic acid-schiff (PAS) staining (C). (D, E) AML-12 cells were transfected with si- Serca2 or si-Control for 72 h, and then subjected to intracellular eukaryotic translation initiation factor 2 subunit α (eIF2α) phosphorylation assessment by Western blotting (D), and quantified (E). n = 3. (F, G) AML-12 cells receiving si- Serca2 or si-Control transfection were incubated with palmitic acid (PA) (400 μM) in the presence or absence of AT-II (50 μM) treatment for 24 h. The cells were collected for Western blotting analysis (F), and quantified (G). n = 3. (H, I) HepG2 cells with farnesoid X receptor ( FXR )-RNAi were cultured with 400 μM of PA and PA + AT-II (50 μM) for 24 h n = 3. The cells were then subjected to Western blotting analysis (H), and quantified (I). (J) Representative images of lipid droplet boron-dipyrromethene (BODIPY) staining in HepG2 cells with or without FXR silencing after 24-h PA (400 μM) and PA + AT-II (50 μM) treatment. Image taken at ×400 magnification. All data were shown as the means ± standard error of the mean (SEM). ∗ P < 0.05; NS: no significance. NC: negative control; AKT: protein kinase B; JNK: c-jun N -terminal kinase; ACTB: actin beta; DAPI: 4′,6-diamidino-2-phenylindole.

Article Snippet: The intracellular Ca 2+ ATPase activity was measured using an Ultramicro Ca 2+ ATPase Diagnostic Kit (Cat# A070-4-2, Nanjing Jiancheng, Nanjing, China).

Techniques: De-Phosphorylation Assay, Injection, Virus, Expressing, Western Blot, Staining, Transfection, Control, Phospho-proteomics, Incubation, Cell Culture, Negative Control

Journal: Advanced Science

Article Title: HUWE1 in Skeletal Muscle Prevents Muscle Fatigue via Maintaining Iron and Calcium Homeostasis

doi: 10.1002/advs.202516719

Figure Lengend Snippet: Pharmacological activation of SERCA or silencing of Sln ameliorates exercise performance in Huwe1 deficient mice. A,B) Time courses of Fura‐2 Ca 2+ signals (F340/F380) in WT and Huwe1 KO C2C12 cells treated by CDN1163. WT and Huwe1 KO C2C12 cells were incubated with/without CDN1163 (10 µM) for 24 h. Afterward s, cells were subjected to recording the baseline of cytoplasmic Ca 2+ by the Fura‐2 imaging approach under the microscope. And then ionomycin was added to monitor the total Ca 2+ storage content. Individual cell trace (gray) and average trace (red) were shown for each group (A); SERCA activity was evaluated (B, n = 3); C–E) WT and Huwe1 cKO mice were intraperitoneally injected with saline or CDN1163 (40 mg kg −1 ) for 7 consecutive days, followed by one bout of exhausted exercise; SERCA activity (C), running time (D) and running distance (E) of each mouse were recorded. n = 5–6 in each group; F–L) Adeno‐associated virus 2/9 (AAV2/9) packaged‐sh Sln and AAV2/9‐EGFP (shNC) were injected into the soleus and gastrocnemius muscles of each leg in WT and Huwe1 cKO mice to knock down Sln, these mice were subjected to one bout of exhausted exercise 4 weeks after injection (n = 3–5 in each group). The expression of Huwe1 (F) and Sln (G) were detected by qPCR; Immunochemistry staining of Sln in indicated groups were shown and hematoxylin staining indicated the nuclei (H, scale bar, 100 µm); SERCA activities were assayed (I); Running time (J), running distance (K) and maximum speed (L) of each mouse in the indicated groups were recorded. Data are presented as Mean ± SEM. Student's t‐test was used to calculate the statistical probability ( p ) values shown between the indicated groups.

Article Snippet: Supernatants were collected after centrifugation at 12 000 g for 10 min. SERCA activity and iron content were measured by using the Ca 2+ ‐ATPase Assay Kit (Nanjing Jiancheng Bioengineering Institute, China) and the Tissue Iron Assay Kit (Nanjing Jiancheng Bioengineering Institute) according to the manufacturer's instructions.

Techniques: Activation Assay, Incubation, Imaging, Microscopy, Activity Assay, Injection, Saline, Virus, Muscles, Knockdown, Expressing, Staining