Journal: Cellular immunology

Article Title: Fluvastatin enhances IL-33-mediated mast cell IL-6 and TNF production.

doi: 10.1016/j.cellimm.2021.104457

Figure Lengend Snippet: Fig. 2. Fluvastatin effects are SCF-dependent. (A) BMMC were cultured in IL-3 ± SCF (50 ng/ml) for three days. Fluvastatin (20 μM) was added for 24 h prior to IL-33 activation for 16 h. Cytokines were measured via ELISA. Data are means ± SEM of 6 populations from 2 independent experiments. (B) BMMC were cultured for 24 h in media containing DMSO (vehicle control) or fluvastatin (20 μM), and surface expression of c-Kit and ST2 was measured by flow cytometry. Data are means ± SD from 17 samples of BMMC populations.

Article Snippet: Fluvastatin sodium salt was purchased from Tocris Bioscience, part of Bio-techne (Minneapolis, MN).

Techniques: Cell Culture, Activation Assay, Enzyme-linked Immunosorbent Assay, Control, Expressing, Flow Cytometry

Journal: Cellular immunology

Article Title: Fluvastatin enhances IL-33-mediated mast cell IL-6 and TNF production.

doi: 10.1016/j.cellimm.2021.104457

Figure Lengend Snippet: Fig. 1. Fluvastatin has opposing effects on IgE and IL-33-mediated cytokine. (A) BMMC were treated with anti-DNP IgE ± 20 μM Fluvastatin for 24 h prior to IgE XL or IL-33 stimulation. Supernatants were collected 16 h after activation and cytokines were measured via ELISA. Data are means ± SEM of 3 populations, representative of 2 inde pendent experiments. (B) BMMC were treated with the indicated dose of fluvastatin for 24 h prior to activation. Cells were then stimulated with IL-33 (100 ng/ml) for 16 h, and cytokines were measured via ELISA. Data are means ± SEM of 9 populations from at least 2 independent experiments. (C) BMMC were treated as described in (A), with RNA collected 4 h after IL-33 stimulation. RT-qPCR was used to measure IL-6 mRNA expression. Data are means ± SEM of 3 populations. (D) Peritoneal mast cells or (E) Skin-derived human mast cells from 3 donors were treated with 20 μM fluvastatin or DMSO for 24 h prior to stimulation with 100 ng/ml of IL-33 stimulation. Supernatants were collected 16 h after activation and cy tokines were measured by ELISA. Data are means ± SEM, with each icon representing a donor.

Article Snippet: Fluvastatin sodium salt was purchased from Tocris Bioscience, part of Bio-techne (Minneapolis, MN).

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Expressing, Derivative Assay

Journal: Cellular immunology

Article Title: Fluvastatin enhances IL-33-mediated mast cell IL-6 and TNF production.

doi: 10.1016/j.cellimm.2021.104457

Figure Lengend Snippet: Fig. 3. TNF blockade does not prevent fluvastatin effects. BMMC were cultured +/- fluvastatin for 24 h as described in Fig. 1. Cells were then activated +/- IL-33 (100 ng/ml) for 16 h in the presence of (A) anti-TNF (5 ng/ml), (B) anti-TNF receptor 1 and 2 (100 ng/ml), or isotype control antibodies. Culture supernatants were analyzed by ELISA. Data shown are means ± SEM from each of 3 BMMC populations analyzed in triplicate.

Article Snippet: Fluvastatin sodium salt was purchased from Tocris Bioscience, part of Bio-techne (Minneapolis, MN).

Techniques: Cell Culture, Control, Enzyme-linked Immunosorbent Assay

Journal: Cellular immunology

Article Title: Fluvastatin enhances IL-33-mediated mast cell IL-6 and TNF production.

doi: 10.1016/j.cellimm.2021.104457

Figure Lengend Snippet: Fig. 4. Fluvastatin-mediated increase in cytokine production depends on inhibiting the isoprenoid arm of the HMG-CoA pathway. (A) A simplified depiction of cholesterol synthesis and the inhibitors used. Dashed arrows indicate summarized pathway steps in which intermediates are not shown. In hibitors are indicated by gray font. (B) BMMC were cultured in vehicle or fluvastatin (20 μM) ± mevalonic acid (1 mM). (C) BMMC were cultured for 24 hrs in the presence of ZA or flu vastatin at 20 μM. (D) BMMC were cultured for 24 hrs in ± fluvastatin (20 μM) in the presence or absence of GGPP, ± FPP at 20 μM. Cultures were then activated with IL-33 (100 ng/ml) for 16 h, and cytokines were measured by ELISA. Data shown are means ± SEM from 6 (B, D) or 14 (C) samples from at least 2 separate experiments.

Article Snippet: Fluvastatin sodium salt was purchased from Tocris Bioscience, part of Bio-techne (Minneapolis, MN).

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Cellular immunology

Article Title: Fluvastatin enhances IL-33-mediated mast cell IL-6 and TNF production.

doi: 10.1016/j.cellimm.2021.104457

Figure Lengend Snippet: Fig. 5. Fluvastatin alters IL-33 induced signaling. (A) BMMC were cultured for 24 hrs in the presence of vehicle or fluvastatin (20 μM) followed by activation with IL- 33 (100 ng/ml) for 0–15 min. Samples were fixed and stained with the indicated antibodies and analyzed via flow cytometry. MFI of phosphoprotein was normalized to vehicle 0′ time point. Data shown are means ± SEM from 3 populations, representative of 2 experiments. (B) BMMC were cultured for 24 h with the indicated chemicals prior to IL-33 stimulation for 16 h. Culture supernatants were analyzed by ELISA. The Akt inhibitors MK-2206 and GDC-0068 were used at 3 μM and 5 μM, respectively. Data shown are means ± SEM from 3 populations. (C) Samples were lysed and analyzed by western blot. Data shown are from 1 of 3 experiments that yielded similar outcomes. (D) BMMC were transfected with vectors encoding luciferase genes from Renilla reniformis under HSV-TK promoter and Firefly under NF- κB response elements. Cultures were then treated vehicle or fluvastatin for 24 h prior to activation IL-33 for 2 h. Ratios of Firefly to Renilla luciferase were normalized to DMSO unstimulated. (E) BMMC were cultured in the presence of vehicle or fluvastatin ± bay11-7082. Cells were activated with IL-33 for 16 h, and cytokines were measured by ELISA. Data are means ± SEM of 9 (D) or 8 (E) populations from 3 independent experiments.

Article Snippet: Fluvastatin sodium salt was purchased from Tocris Bioscience, part of Bio-techne (Minneapolis, MN).

Techniques: Cell Culture, Activation Assay, Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Western Blot, Transfection, Luciferase

Journal: Cellular immunology

Article Title: Fluvastatin enhances IL-33-mediated mast cell IL-6 and TNF production.

doi: 10.1016/j.cellimm.2021.104457

Figure Lengend Snippet: Fig. 6. Fluvastatin enhances IL-33-mediated cytokine production and neutrophil recruitment in vivo. (A) C57BL/6J mice were injected i.p. with vehicle or fluvastatin (0.5 mg/mouse) 24 h and again 1 h prior to IL-33 (1 µg/mouse) or PBS. Schematic created with BioRender. Peritoneal lavage and cardiac puncture were performed 4 h after IL-33 injection. (B) Cells from the peritoneal lavage were stained with anti-CD45 and the percentage positive cells was used to calculate total cell numbers in peritoneal lavage fluid. (C) The indicated innate immune lineages were assessed by flow cytometry. (D) T cell and B cell populations were assessed by flow cytometry. CD11b, Ly6C, and Ly6G and analyzed using flow cytometry. (E) Plasma and peritoneal lavage fluid were collected and assessed for IL-6 via ELISA. Data are means ± SEM of n = 5 mice, analyzed by ANOVA, and representative of 3 independent experiments.

Article Snippet: Fluvastatin sodium salt was purchased from Tocris Bioscience, part of Bio-techne (Minneapolis, MN).

Techniques: In Vivo, Injection, Staining, Flow Cytometry, Clinical Proteomics, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Highly variable biological effects of statins on cancer, non-cancer, and stem cells in vitro

doi: 10.1038/s41598-024-62615-w

Figure Lengend Snippet: Effect of statins on size and compactness of spheroids. ( a ) ADMSC stem cells, ( b ) pancreatic cancer MiaPaCa-2 cells, concentration of statins—20 µM, Ctr—methanol treated spheroids, P—pravastatin, R—rosuvastatin, L—lovastatin, F—fluvastatin, A—atorvastatin, Pi—pitavastatin, C—cerivastatin, S—simvastatin. Statins were added once, after spheroid formation, 10 weeks ( a ) or 3.5 weeks ( b ) after inoculation. Experiment was carried out in biological dodecaplicates.

Article Snippet: In all experiments, pure forms (≥ 98%) of the following statins were used: atorvastatin (A7658), lovastatin (M1687), simvastatin (S3449), fluvastatin (F4482), cerivastatin (C1668), pravastatin (P6801), rosuvastatin (R5974), and pitavastatin (P3576) (LKT Laboratories, USA).

Techniques: Concentration Assay

Journal: Scientific Reports

Article Title: Highly variable biological effects of statins on cancer, non-cancer, and stem cells in vitro

doi: 10.1038/s41598-024-62615-w

Figure Lengend Snippet: Effect of statins on the spheroid formation. ( a ) ADMSC stem cells, ( b ) pancreatic cancer MiaPaCa-2 cells, concentration of statins—20 µM, Ctr methanol treated spheroids, P —pravastatin, R —rosuvastatin, L —lovastatin, F —fluvastatin, A —atorvastatin, Pi —pitavastatin, C —cerivastatin, S —simvastatin. Statins were added once, 24 h after cell inoculation. Experiment was carried out in biological dodecaplicates.

Article Snippet: In all experiments, pure forms (≥ 98%) of the following statins were used: atorvastatin (A7658), lovastatin (M1687), simvastatin (S3449), fluvastatin (F4482), cerivastatin (C1668), pravastatin (P6801), rosuvastatin (R5974), and pitavastatin (P3576) (LKT Laboratories, USA).

Techniques: Concentration Assay

Journal: FEBS Open Bio

Article Title: Statins induce monocytic differentiation in acute myeloid leukemia cells through the KLF4 / DPYSL2A axis

doi: 10.1002/2211-5463.70104

Figure Lengend Snippet: Statins induced monocytic differentiation and suppressed cell proliferation in THP‐1 cells by stimulating the KLF4/DPYSL2A axis. (A) Dose–response curves of the five statins in THP‐1 cells, along with the corresponding IC 50 values for each statin. The cells were treated with various statin concentrations for 48 h ( n = 3). (B) Growth inhibition following statin treatment of THP‐1 cells. To assess cell proliferation, 1 × 10 4 cells were seeded in a six‐well plate and cultured with DMSO or statins (2 or 4 μ m ). Trypan blue dye exclusion assays were performed after 6 days of treatment ( n = 4). Abbreviations: Ator, atorvastatin; Flu, fluvastatin; Lo, lovastatin; Me, mevastatin; Pra, pravastatin. (C) Relative mRNA expression levels of KLF4 and DPYSL2A in THP‐1 cells treated as described in (B). Total RNA was then extracted and analyzed by RT‐qPCR. Values were normalized to the GAPDH expression levels ( n = 3). (D) Cell surface expression levels of CD11b and CD14 in THP‐1 cells treated as described in (B) ( n = 3). (E) Morphological changes in THP‐1 cells treated with DMSO or statins (4 μ m ) for 6 days. (F) Apoptosis of THP‐1 cells treated as described in (B). The annexin V‐positive cells were scored using flow cytometric analysis ( n = 3). Data are presented as the mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001; two‐tailed Student's t ‐test.

Article Snippet: Atorvastatin (HY‐B0589), fluvastatin sodium (HY‐14664A), lovastatin (HY‐N0504), mevastatin (HY‐17408), pravastatin sodium (HY‐B0165A), and FTI‐277 (HY‐15872A) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Inhibition, Cell Culture, Expressing, Quantitative RT-PCR, Two Tailed Test

Journal: FEBS Open Bio

Article Title: Statins induce monocytic differentiation in acute myeloid leukemia cells through the KLF4 / DPYSL2A axis

doi: 10.1002/2211-5463.70104

Figure Lengend Snippet: Fluvastatin induces monocytic differentiation in HL‐60 cells via the KLF4/DPYSL2A axis. (A) Growth inhibition by fluvastatin in HL‐60 cells. To assess cell proliferation, 1 × 10 4 cells were seeded in a six‐well plate and cultured with DMSO or fluvastatin (2 or 4 μ m ). Trypan blue dye exclusion assays were performed after 6 days of treatment ( n = 3). (B) Relative mRNA expression levels of KLF4 and DPYSL2A in HL‐60 cells treated with DMSO or 4 μ m fluvastatin. Total RNA was then extracted and analyzed by RT‐qPCR. Values were normalized to the GAPDH expression levels ( n = 3). (C) Cell surface expression levels of CD11b and CD14 in HL‐60 cells treated as described in (B) ( n = 3). (D) Apoptosis in HL‐60 cells treated as described in (B). The annexin V‐positive cells were scored using flow cytometric analysis ( n = 3). Data are presented as the mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001; two‐tailed Student's t ‐test was used.

Article Snippet: Atorvastatin (HY‐B0589), fluvastatin sodium (HY‐14664A), lovastatin (HY‐N0504), mevastatin (HY‐17408), pravastatin sodium (HY‐B0165A), and FTI‐277 (HY‐15872A) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Inhibition, Cell Culture, Expressing, Quantitative RT-PCR, Two Tailed Test

Journal: FEBS Open Bio

Article Title: Statins induce monocytic differentiation in acute myeloid leukemia cells through the KLF4 / DPYSL2A axis

doi: 10.1002/2211-5463.70104

Figure Lengend Snippet: Monocytic differentiation induced by fluvastatin depends on the inhibition of the MVA pathway. (A) Relative mRNA expression levels of KLF4 in THP‐1 cells. The cells were treated with DMSO, 4 μ m fluvastatin alone, or 4 μ m fluvastatin in combination with 100 μ m MVA for 6 days. Total RNA was then extracted and analyzed by RT‐qPCR. Values were normalized to the GAPDH expression levels ( n = 3). (B) Relative mRNA expression levels of DPYSL2A in THP‐1 cells. The cells were treated as described in (A), and then total RNA was extracted and analyzed by RT‐qPCR. Values were normalized to the GAPDH expression levels ( n = 3). (C) Cell surface expression levels of CD11b in THP‐1 cells. The cells were treated as described in (A) and then harvested for flow cytometric analysis ( n = 5). (D) Cell surface expression levels of CD14 in THP‐1 cells. The cells were treated as described in (A) and then harvested for flow cytometric analysis ( n = 5). (E) Rescue of THP‐1 cells from fluvastatin‐induced apoptosis following MVA treatment. The cells were treated as described in (A). The annexin V‐positive cells were scored using flow cytometric analysis ( n = 5). (F) Rescue of THP1 cells from fluvastatin following MVA treatment. To assess cell proliferation, 1 × 10 4 cells were seeded in a six‐well plate and treated as described in (A). Trypan blue dye exclusion assays were performed every other day ( n = 5). Data are presented as the mean ± SEM. * P < 0.05, ** P < 0.01, N.S., not significant; one‐way analysis of variance (ANOVA), followed by Tukey's post hoc test, was used.

Article Snippet: Atorvastatin (HY‐B0589), fluvastatin sodium (HY‐14664A), lovastatin (HY‐N0504), mevastatin (HY‐17408), pravastatin sodium (HY‐B0165A), and FTI‐277 (HY‐15872A) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Inhibition, Expressing, Quantitative RT-PCR

Journal: Drug metabolism and disposition: the biological fate of chemicals

Article Title: Comparative Hepatic and Intestinal Efflux Transport of Statins.

doi: 10.1124/dmd.121.000430

Figure Lengend Snippet: Fig. 3. Concentration-dependent transport of fluvastatin in BCRP, MRP2, MRP3, MRP4, MRP8, P-gp, and control vesicles. The time of incubation and vesicle amount were 5 minutes and 7.5 mg, respectively. Results are presented as mean ± S.D. transport, which is calculated from the means of each separate experiment. In total, three separate experiments were performed for each transporter with triplicate samples in each experiment.

Article Snippet: Racemic fluvastatin, 3R,5S-fluvastatin, 3S,5R-fluvastatin, and pitavastatin were obtained from Santa Cruz Biotechnology (Dallas, TX).

Techniques: Concentration Assay, Control, Incubation