Journal: Journal of enzyme inhibition and medicinal chemistry

Article Title: Synthesis and evaluation of 5, 6-dihydro-8 H -isoquinolino[1, 2- b ]quinazolin-8-one derivatives as novel non-lipogenic ABCA1 up-regulators with inhibitory effects on macrophage-derived foam cell formation.

doi: 10.1080/14756366.2025.2470310

Figure Lengend Snippet: Figure 1. (A) The chemical structure of LXR agonist T0901317; (B) The chemical structures of quinazolinone-containing compounds with ABCA1 up-regulating activity and the chemical structure of 5, 6-dihydro-8H-isoquinolino[1, 2-b]quinazolin-8-one in the present study.

Article Snippet: Mouse-derived macrophage RaW264.7, human cervical cancer cell hela and human hepatocellular carcinoma cell hepG2 were obtained from the typical culture collection centre of china. the human oxidised low density lipoprotein (ox-lDl) and Dii-ox-lDl (ox-lDl labelled with 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate) were purchased from Yiyuan Biotechnologies (GuangZhou, china). the rabbit polyclonal aBca1 antibody was obtained from Novus Biological inc. (Usa). the anti-β-actin antibody and the secondary antibodies (anti-rabbit and anti-mouse igG-hRP) were obtained from santa cruz Biotech corp (Usa).

Techniques: Activity Assay

Journal: Journal of enzyme inhibition and medicinal chemistry

Article Title: Synthesis and evaluation of 5, 6-dihydro-8 H -isoquinolino[1, 2- b ]quinazolin-8-one derivatives as novel non-lipogenic ABCA1 up-regulators with inhibitory effects on macrophage-derived foam cell formation.

doi: 10.1080/14756366.2025.2470310

Figure Lengend Snippet: Figure 2. The SAR of 5, 6-dihydro-8H-isoquinolino[1, 2-b]quinazolin-8-one compounds on ABCA1 promoter activation.

Article Snippet: Mouse-derived macrophage RaW264.7, human cervical cancer cell hela and human hepatocellular carcinoma cell hepG2 were obtained from the typical culture collection centre of china. the human oxidised low density lipoprotein (ox-lDl) and Dii-ox-lDl (ox-lDl labelled with 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate) were purchased from Yiyuan Biotechnologies (GuangZhou, china). the rabbit polyclonal aBca1 antibody was obtained from Novus Biological inc. (Usa). the anti-β-actin antibody and the secondary antibodies (anti-rabbit and anti-mouse igG-hRP) were obtained from santa cruz Biotech corp (Usa).

Techniques: Activation Assay

Journal: Journal of enzyme inhibition and medicinal chemistry

Article Title: Synthesis and evaluation of 5, 6-dihydro-8 H -isoquinolino[1, 2- b ]quinazolin-8-one derivatives as novel non-lipogenic ABCA1 up-regulators with inhibitory effects on macrophage-derived foam cell formation.

doi: 10.1080/14756366.2025.2470310

Figure Lengend Snippet: Figure 3. Effects of compound 3 on the expression of ABCA1 in Hela and RAW264.7 cells. (A), Dose-dependent effects on the activation of ABCA1 promoter upon treatment with 3 in Hela cells; (B), Dose-dependent effects of compound 3 on the mRNA expression of ABCA1 in RAW264.7 cells; (C), Time-dependent effects of compound 3 on the protein expression of ABCA1 in RAW264.7 cells after treatment with compound 3 at 10 μM for the indicated time; (D) Dose-dependent effects of compound 3 on the protein expression of ABCA1 in RAW264.7 cells after treatment with compound 3 at various concentrations for 24 h. Bands from time-dependent blots and dose-response blots were quantified by densitometry, normalised to β-actin, and expressed as fold of vehicle treatment. Data are represented as mean ± SD from at least two independent experiments. Veh, 0.1% DMSO. Significance: *p < 0.05, **p < 0.01, ****< 0.0001, vs. vehicle group.

Article Snippet: Mouse-derived macrophage RaW264.7, human cervical cancer cell hela and human hepatocellular carcinoma cell hepG2 were obtained from the typical culture collection centre of china. the human oxidised low density lipoprotein (ox-lDl) and Dii-ox-lDl (ox-lDl labelled with 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate) were purchased from Yiyuan Biotechnologies (GuangZhou, china). the rabbit polyclonal aBca1 antibody was obtained from Novus Biological inc. (Usa). the anti-β-actin antibody and the secondary antibodies (anti-rabbit and anti-mouse igG-hRP) were obtained from santa cruz Biotech corp (Usa).

Techniques: Expressing, Activation Assay

Journal: Journal of enzyme inhibition and medicinal chemistry

Article Title: Synthesis and evaluation of 5, 6-dihydro-8 H -isoquinolino[1, 2- b ]quinazolin-8-one derivatives as novel non-lipogenic ABCA1 up-regulators with inhibitory effects on macrophage-derived foam cell formation.

doi: 10.1080/14756366.2025.2470310

Figure Lengend Snippet: Figure 4. Suppressing effects of GSK2033 on the up-regulation of ABCA1 induced by compound 3. (A) Suppressing effects on the activation of ABCA1 promoter upon treatment with T0901317 (0.1 μM), 3 (10 μM), GSK2033 (1 μM) and 3 together with GSK2033 in Hela cells; (B) Suppressing effects on the expression of ABCA1 protein upon treatment with T0901317 (0.1 μM), 3 (10 μM), GSK2033 (1 μM) and 3 together with GSK2033 in RAW264.7 cells. Bands from western blots were quantified by densitometry, normalised to β-actin, and expressed as fold of vehicle treatment. Data are represented as mean ± SD from at least two independent experiments. Veh, 0.1% DMSO; T0, T0901317; GSK, GSK2033. Significance: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, vs. vehicle group; #p < 0.01, vs. compound 3 treated-group.

Article Snippet: Mouse-derived macrophage RaW264.7, human cervical cancer cell hela and human hepatocellular carcinoma cell hepG2 were obtained from the typical culture collection centre of china. the human oxidised low density lipoprotein (ox-lDl) and Dii-ox-lDl (ox-lDl labelled with 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate) were purchased from Yiyuan Biotechnologies (GuangZhou, china). the rabbit polyclonal aBca1 antibody was obtained from Novus Biological inc. (Usa). the anti-β-actin antibody and the secondary antibodies (anti-rabbit and anti-mouse igG-hRP) were obtained from santa cruz Biotech corp (Usa).

Techniques: Activation Assay, Expressing, Western Blot

Journal: Biomedicines

Article Title: Small GTPase ARL4C Associated with Various Cancers Affects Microtubule Nucleation.

doi: 10.3390/biomedicines12122872

Figure Lengend Snippet: Figure 2. Treatment using LXR/RXR ligands does not lead to an increase in the endogenous ABCA1 or ARL4C level in many cell lines: (A) Titration of BS-C-1 lysate demonstrates that the ratio of levels of ARL4C in Vero and BS-C-1 cells is 1:4.5. (B) The level of ARL4C is not increased in both Vero and BS-C-1 cells even after 7 days of treatment with T0901317 and bexarotene. (C) LXR/RXR-dependent pathway is activated in HeLa cells and weakly activated in MCF7 cells after 48h exposure to 2.5 µM T0901317/bexarotene; however, not in Vero cells even after prolonged treatment. (D) Neither ABCA1 nor ARL4C levels increase after treatment of COS7 and U2OS cells with RXR/LXR ligands. Long-term treatment of the MCF7 cells results in only a slight increase in the ARL4C level.

Article Snippet: Then, the membrane was blocked with 5% skimmed milk in TBST for 1 h. Rabbit polyclonal antibody against ARL4C (Novus Biologicals, Centennial, CO, USA), mouse monoclonal antibody against GAPDH (ThermoFisher Scientific, Waltham, MA, USA), rabbit polyclonal antibody against ABCA1 (Novus Biologicals, Centennial, CO, USA), mouse monoclonal antibody against α-tubulin, and clone DM1A (Santa Cruz Biotechnology, Dallas, TX, USA) were used as primary antibodies.

Techniques: Titration

Journal: Acta Pharmaceutica Sinica. B

Article Title: PPAR α affects hepatic lipid homeostasis by perturbing necroptosis signals in the intestinal epithelium

doi: 10.1016/j.apsb.2024.08.021

Figure Lengend Snippet: Pparα deficiency in intestinal epithelium increases the translocation of gut-derived antigens into the liver. (A) Intestinal permeability assessment (FITC-dextran, 4 kD) in 8-week-old mice ( n = 10). (B) Relative mRNA levels of Zo-1 and Cldn8 in the ileum from 8-week-old mice ( n = 5). (C) The relative proportion of bacterial species in the cecum content by 16S rRNA gene sequencing ( n = 6). (D) Bugbase phenotypic prediction of gut microbiota in 8-week-old mice ( n = 6). (E) The mRNA and protein levels of PV1 in the ileum of 8-week-old mice ( n = 5). (F) Transmission electron microscopy images of the diaphragm (red star) in the capillaries from ileum sections in 24-week-old mice ( n = 3). (G) Representative images of fluorescence microscopy and transmission electron in 8-week-old mice treated with FITC-LPS (green) or Au-LPS ( n = 3–5). (H) Portal HDL-C levels in 8-week-old mice ( n = 10). (I) The protein levels of APOA1 and ABCA1 in the ileum of 8-week-old mice ( n = 5). (J) Relative mRNA levels of Apoa1 , Pon1 , and Pon3 in the ileum of 8-week-old mice ( n = 5). (K) Serum APOA1 levels in 8-, 16- and 32-week-old mice ( n = 8–10). (L) Serum APOA1 levels in 8-week-old mice exposed to HFCS for 14 days ( n = 8). (M) Serum APOA1 levels in 16-week-old Pparα Δhep mice ( n = 10). (N) Schematic representation of a cocktail of broad-spectrum antibiotics (Abx) experimental design. (O) Representative images stained with H&E and Oil Red O, and immunofluorescent staining for F4/80 (red) in the liver from 8-week-old mice treated with Abx ( n = 5). (P) Triglyceride in serum and liver treated with Abx ( n = 10). (Q) Relative mRNA levels of genes related to triglyceride accumulation in the liver from 8-week-old mice treated with Abx ( n = 5). (R) Hepatic levels of cytokines from 8-week-old mice treated with Abx ( n = 5). (S) Protein levels of F4/80 in the liver of 8-week-old mice treated with Abx ( n = 3). (T) Relative mRNA levels of F4/80, Clec4f , and Cd14 in the liver of 8-week-old mice treated with Abx ( n = 5). LD: lipid droplet; M: mitochondria; FITC-LPS: fluorescein isothiocyanate (FITC)-LPS; Au-LPS: LPS-gold-complexes. Data are shown as the mean ± SD. An unpaired two-tailed Student's t -test; ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: These samples were incubated overnight with a polyclonal rabbit anti-mouse F4/80 (1:1000, #A1256, ABclonal), polyclonal rabbit anti-mouse APOA1 (1:2000, #A14211, ABclonal), polyclonal rabbit anti-mouse ABCA1 (1:1000, #NB400-105, Novus Biological, Centennial), monoclonal rabbit anti-mouse CD14 (1:1000, #A19011, ABclonal), or polyclonal rabbit anti-mouse PV1 (1:2000, #A15906, ABclonal).

Techniques: Translocation Assay, Derivative Assay, Permeability, Sequencing, Transmission Assay, Electron Microscopy, Fluorescence, Microscopy, Staining, Two Tailed Test

Journal: iScience

Article Title: Alphaherpesvirus manipulates retinoic acid metabolism for optimal replication

doi: 10.1016/j.isci.2024.110144

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-ABCA1 , Cell Signaling Technology , Cat# 96292.

Techniques: Recombinant, Transfection, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Software

Journal: Chinese Medicine

Article Title: Combination of mangiferin and T0901317 targeting autophagy promotes cholesterol efflux from macrophage foam cell in atherosclerosis

doi: 10.1186/s13020-023-00876-9

Figure Lengend Snippet: Treatment of T0 and MGF activates expression of LXR target molecules in macrophage and inhibits foam cell formation. A Peritoneal macrophages collected from mice were stained with Oil Red O or anti-bodipy antibody (green) to evaluate formation of foam cells (> 10 lipid droplets per cell, > 10 fields per sample), scale bar, 20 μm; and cholesterol contents were measured. *** p < 0.001, n = 3. Expression of ( B ) ABCA1 and ( C ) ABCG1 (stained with red fluorescent color) was determined by immunofluorescent staining of aortic root cross sections from Apoe −/− mice and quantified by a computer-assisted image analysis protocol. Scale bar, 100 μm. ** p < 0.01, n = 3. Expression of ABCA1, ABCG1, LXRα and CD36 in cultured peritoneal macrophages ( D ) and RAW264.7 cells ( E ) was determined by western blot with total proteins extracted from cell samples. *** p < 0.001, ** p < 0.01, * p < 0.05 (n = 3)

Article Snippet: Primary rabbit polyclonal antibodies against ABCA1 (ab18180), ABCG1 (ab218528), LC3 (ab192890), SREBP-1c (ab28481), FAS (ab133619), and β-actin (ab8227) were purchased from Abcam (Cambridge, MA, USA).

Techniques: Expressing, Staining, Cell Culture, Western Blot

Journal: Chinese Medicine

Article Title: Combination of mangiferin and T0901317 targeting autophagy promotes cholesterol efflux from macrophage foam cell in atherosclerosis

doi: 10.1186/s13020-023-00876-9

Figure Lengend Snippet: Model for the function of combined treatment of T0 and MGF in the AS related dyslipidemia targeting autophagy in cholesterol efflux from macrophage foam cells. On the one hand, T0 alone or T0 and MGF promote macrophage cholesterol efflux by activating LXRα to augment the expression of ABCA1 and ABCG1 and also enhance lipophagy in atherosclerotic lesion. Of note, in mammalian macrophage foam cells, lipid droplets are tagged for autophagic fusion, possibly beginning with mTOR-AMPK signaling to initiate lipid droplet degradation for further cholesterol depletion. On the other hand, T0-induced hepatic lipid abnormal accumulation is attenuated by MGF. In this scenario, MGF may activate AMPK signaling to suppress lipid synthesis and accelerate lipolysis for β-oxidation of free fatty acid. Alternatively, hepatic lipid droplets may be degraded through autophagy mechanism mediated by AMPK, and relative classical factors LC3, ATGs and p38 to facilitate hepatic lipophagy

Article Snippet: Primary rabbit polyclonal antibodies against ABCA1 (ab18180), ABCG1 (ab218528), LC3 (ab192890), SREBP-1c (ab28481), FAS (ab133619), and β-actin (ab8227) were purchased from Abcam (Cambridge, MA, USA).

Techniques: Expressing