Journal: The American Journal of Pathology

Article Title: AP39, a Modulator of Mitochondrial Bioenergetics, Reduces Antiangiogenic Response and Oxidative Stress in Hypoxia-Exposed Trophoblasts

doi: 10.1016/j.ajpath.2018.09.007

Figure Lengend Snippet: AP39 pretreatment reduces production of mitochondrial-derived reactive oxygen species and preserves active mitochondria in hypoxia-exposed primary trophoblasts. Immunofluorescence analysis of MitoTracker Green (active mitochondria; A–D) and MitoSOX Red (superoxide production; F–I) in primary trophoblasts treated with 50 μmol/L AP39 for 30 minutes, then exposed to 18 hours of 5% hypoxia. Quantitation of MitoTracker Green (E) and MitoSOX Red (J) immunofluorescence in trophoblasts: OD per area (pixels2) of cell surface area was calculated in four high-power fields per sample. n = 4 per group (E and J). ∗P < 0.05 by Kruskal Wallis test. Scale bar = 50 μm (A–D and F–I).

Article Snippet: AP39 (10-oxo-10-[4-(3-thioxo-3H-1,2-dithiol-5yl)phenoxy]decyl triphenyl-phosphonium bromide) was obtained from Cayman Chemical (Ann Arbor, MI).

Techniques: Derivative Assay, Immunofluorescence, Quantitation Assay

Journal: The American Journal of Pathology

Article Title: AP39, a Modulator of Mitochondrial Bioenergetics, Reduces Antiangiogenic Response and Oxidative Stress in Hypoxia-Exposed Trophoblasts

doi: 10.1016/j.ajpath.2018.09.007

Figure Lengend Snippet: AP39 pretreatment reduces hypoxia-inducible factor (HIF)-1α and sFLT1 protein expression in hypoxia-exposed human primary trophoblasts and maintains cell viability. A: Representative Western blot analysis of HIF-1α (one of three experiments) in primary trophoblasts pretreated with AP39 (10, 25, and 50 μmol/L) for 30 minutes, then exposed to normoxia (21% O2) or 18 hours of 5% hypoxia. B: Summary densitometry of three HIF-1α Western blot analyses. C: sFLT1 enzyme-linked immunosorbent assay in primary trophoblasts with same treatment as in A and B. D: Cell viability in primary trophoblasts assessed by Dojindo's Cell Counting Kit-8 colorimetric assay. ∗P < 0.05 compared with control; ∗∗P < 0.01 compared with control by Kruskal-Wallis test, followed by Dunn's multiple comparison post hoc test.

Article Snippet: AP39 (10-oxo-10-[4-(3-thioxo-3H-1,2-dithiol-5yl)phenoxy]decyl triphenyl-phosphonium bromide) was obtained from Cayman Chemical (Ann Arbor, MI).

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Cell Counting, Colorimetric Assay, Control, Comparison

Journal: The American Journal of Pathology

Article Title: AP39, a Modulator of Mitochondrial Bioenergetics, Reduces Antiangiogenic Response and Oxidative Stress in Hypoxia-Exposed Trophoblasts

doi: 10.1016/j.ajpath.2018.09.007

Figure Lengend Snippet: AP39 pretreatment induces mitochondrial COX IV activity and expression in hypoxia-exposed human primary trophoblasts. Representative COX enzyme histochemistry and summary data (A and B) and representative Western blot analysis of COX IV and summary data (C and D) in primary trophoblasts pretreated with AP39 (10, 25, and 50 μmol/L) for 30 minutes, then exposed to normoxia or 18 hours of 5% hypoxia. Quantitation is shown. ∗P < 0.05 compared with control by Kruskal Wallis test, followed by Dunn's multiple comparison post hoc test. Scale bar = 50 μm (A).

Article Snippet: AP39 (10-oxo-10-[4-(3-thioxo-3H-1,2-dithiol-5yl)phenoxy]decyl triphenyl-phosphonium bromide) was obtained from Cayman Chemical (Ann Arbor, MI).

Techniques: Activity Assay, Expressing, Western Blot, Quantitation Assay, Control, Comparison

Journal: The American Journal of Pathology

Article Title: AP39, a Modulator of Mitochondrial Bioenergetics, Reduces Antiangiogenic Response and Oxidative Stress in Hypoxia-Exposed Trophoblasts

doi: 10.1016/j.ajpath.2018.09.007

Figure Lengend Snippet: Hydrogen sulfide generation is detected in AP39-treated human primary trophoblast cells. Representative immunofluorescence assay for 7-azido-4-methylcoumarin (AzMC; green fluorescence; top and bottom left panels) and corresponding bright field images (top and bottom right panels) were performed in 50 μmol/L AP39-treated (bottom left and right panels) or nontreated (top left and right panels) human primary trophoblast cells. Scale bar = 25 μm.

Article Snippet: AP39 (10-oxo-10-[4-(3-thioxo-3H-1,2-dithiol-5yl)phenoxy]decyl triphenyl-phosphonium bromide) was obtained from Cayman Chemical (Ann Arbor, MI).

Techniques: Immunofluorescence, Fluorescence

Journal: Oxidative Medicine and Cellular Longevity

Article Title: AP39, a Mitochondria-Targeted Hydrogen Sulfide Donor, Supports Cellular Bioenergetics and Protects against Alzheimer's Disease by Preserving Mitochondrial Function in APP/PS1 Mice and Neurons

doi: 10.1155/2016/8360738

Figure Lengend Snippet: AP39 generates H 2 S in WT neurons, primarily in the mitochondria. (a) The contribution of AP39 to H 2 S production in neurons. Neurons from WT mice were treated with various concentrations of AP39 for 2 h, and H 2 S production was detected by methylene blue assay. (b) Neurons were treated with different concentrations of AP39 for 2 h, and intracellular H 2 S was detected using the fluorescent probe AzMC. DAPI was used to stain nuclei, and MitoTracker was used to stain mitochondria. The colocalization of H 2 S with mitochondria was indicated by the overlapping of red (mitochondria) and green (H 2 S) fluorescence in the merged image. Note the concentration-dependent increase in the H 2 S signal in response to AP39 treatment. ∗ P < 0.05, ∗∗ P < 0.01 compared with control treatment (no AP39).

Article Snippet: AP39, a novel mitochondria-targeted H 2 S donor, was designed and synthesized by Medicilon Inc., as described previously [ , ].

Techniques: Staining, Fluorescence, Concentration Assay, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: AP39, a Mitochondria-Targeted Hydrogen Sulfide Donor, Supports Cellular Bioenergetics and Protects against Alzheimer's Disease by Preserving Mitochondrial Function in APP/PS1 Mice and Neurons

doi: 10.1155/2016/8360738

Figure Lengend Snippet: Biphasic effects of AP39 on the cellular bioenergetics of the WT neurons. Neurons from WT mice were incubated in AP39 (25-250 nM) for 2 h, and bioenergetic parameters were measured using an Extracellular Flux Analyzer. (a) Representative tracings. (b) The calculated bioenergetic parameters. # and ## indicate a significant enhancement in the bioenergetic parameter, compared to the control group (no AP39) ( P < 0.05 and P < 0.01, resp.); ∗ indicates a significant reduction in the bioenergetic parameter, compared to the control group ( P < 0.05).

Article Snippet: AP39, a novel mitochondria-targeted H 2 S donor, was designed and synthesized by Medicilon Inc., as described previously [ , ].

Techniques: Incubation, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: AP39, a Mitochondria-Targeted Hydrogen Sulfide Donor, Supports Cellular Bioenergetics and Protects against Alzheimer's Disease by Preserving Mitochondrial Function in APP/PS1 Mice and Neurons

doi: 10.1155/2016/8360738

Figure Lengend Snippet: Cytoprotective effects of AP39 in APP/PS1 neurons. (a) A time course shows that increasing levels of A β 42 are released into the culture media of neurons from transgenic mice but not their WT littermates. (b) A β was overexpressed in neurons from APP/PS1 mice compared to A β expression in neurons from their WT littermates. (c) Representative of A β blots is shown with quantification. (d) The effects of AP39 (25–250 nM) treatment for 24 h on cell viability in WT neurons. AP39 alone did not affect MTT conversion. (e) The effects of AP39 (25–250 nM) on MTT conversion in APP/PS1 neurons. There was a decrease in MTT conversion in the APP/PS1 neurons compared to the WT neurons; these effects were attenuated by AP39. (f) After treatment of WT neurons with AP39 for 24 h, AP39 alone did not affect LDH release in the cellular medium. (g) The effects of AP39 on LDH release from APP/PS1 neurons. ∗ P < 0.05, versus the APP/PS1 group; # P < 0.05, the APP/PS1 group versus the WT group.

Article Snippet: AP39, a novel mitochondria-targeted H 2 S donor, was designed and synthesized by Medicilon Inc., as described previously [ , ].

Techniques: Transgenic Assay, Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: AP39, a Mitochondria-Targeted Hydrogen Sulfide Donor, Supports Cellular Bioenergetics and Protects against Alzheimer's Disease by Preserving Mitochondrial Function in APP/PS1 Mice and Neurons

doi: 10.1155/2016/8360738

Figure Lengend Snippet: Protective effects of AP39 on the cellular bioenergetics of APP/PS1 neurons. APP/PS1 neurons were incubated in AP39 (100 nM) for 24 h, and bioenergetic parameters were measured using the Extracellular Flux Analyzer. (a) Representative tracings are shown. (b) The calculated bioenergetic parameters are shown. ∗ indicates a significant enhancement of the bioenergetic parameter compared to the control (H 2 O) ( P < 0.05); # indicates a significant reduction in the bioenergetic parameter compared to the control (WT + H 2 O) ( P < 0.05).

Article Snippet: AP39, a novel mitochondria-targeted H 2 S donor, was designed and synthesized by Medicilon Inc., as described previously [ , ].

Techniques: Incubation, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: AP39, a Mitochondria-Targeted Hydrogen Sulfide Donor, Supports Cellular Bioenergetics and Protects against Alzheimer's Disease by Preserving Mitochondrial Function in APP/PS1 Mice and Neurons

doi: 10.1155/2016/8360738

Figure Lengend Snippet: AP39 protected mitochondrial function in APP/PS1 neurons. (a) AP39 increased the cellular ATP levels. (b) AP39 protected mtDNA integrity. (c) ROS levels were detected by flow cytometry. (d) AP39 reduced ROS levels. ## P < 0.01, compared with the WT group; ∗∗ P < 0.01, compared with the control group (H 2 O).

Article Snippet: AP39, a novel mitochondria-targeted H 2 S donor, was designed and synthesized by Medicilon Inc., as described previously [ , ].

Techniques: Flow Cytometry, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: AP39, a Mitochondria-Targeted Hydrogen Sulfide Donor, Supports Cellular Bioenergetics and Protects against Alzheimer's Disease by Preserving Mitochondrial Function in APP/PS1 Mice and Neurons

doi: 10.1155/2016/8360738

Figure Lengend Snippet: AP39 reduced the expression of proteins involved in mitochondrial fusion but increased the expression of a mitochondrial fission protein. (a) Protein samples were probed for Drp1, Fis1, Mfn1, Mfn2, OPA-1, and VDAC expression. (b–f) Representative blots are shown with quantification. ## P < 0.01 AD mice receiving water compared to WT mice receiving water; ∗∗ P < 0.01 mice receiving AP39 compared with mice receiving water.

Article Snippet: AP39, a novel mitochondria-targeted H 2 S donor, was designed and synthesized by Medicilon Inc., as described previously [ , ].

Techniques: Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: AP39, a Mitochondria-Targeted Hydrogen Sulfide Donor, Supports Cellular Bioenergetics and Protects against Alzheimer's Disease by Preserving Mitochondrial Function in APP/PS1 Mice and Neurons

doi: 10.1155/2016/8360738

Figure Lengend Snippet: AP39 generates H 2 S in WT mice and AP39 increased H 2 S levels in APP/PS1 mice. (a) The contribution of AP39 to H 2 S production in the cortex and hippocampus of WT mice. (b) AP39 increased H 2 S levels in APP/PS1 mice. ## P < 0.01, compared with the WT group; ∗∗ P < 0.01, compared with the control group (H 2 O).

Article Snippet: AP39, a novel mitochondria-targeted H 2 S donor, was designed and synthesized by Medicilon Inc., as described previously [ , ].

Techniques: Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: AP39, a Mitochondria-Targeted Hydrogen Sulfide Donor, Supports Cellular Bioenergetics and Protects against Alzheimer's Disease by Preserving Mitochondrial Function in APP/PS1 Mice and Neurons

doi: 10.1155/2016/8360738

Figure Lengend Snippet: AP39 ameliorated the learning and memory deficits of APP/PS1 mice. WT or AD mice were treated with water or AP39 (100 nM/kg) for 6 weeks, followed by assessment using the Morris water maze and novel object recognition task tests. (a) Spatial learning and memory in AD mice are scored as the latency to locate a hidden platform. (b) After 24 h, a 60-s probe trial was performed. (c) Spatial learning was tested as the latency to locate a hidden platform for the 25 nM/kg AP39-treated mice and scored. (d) Effects of AP39 on the memory performance of 12-month WT or AD mice treated with water or AP39 were tested in the NORT test.

Article Snippet: AP39, a novel mitochondria-targeted H 2 S donor, was designed and synthesized by Medicilon Inc., as described previously [ , ].

Techniques:

Journal: Oxidative Medicine and Cellular Longevity

Article Title: AP39, a Mitochondria-Targeted Hydrogen Sulfide Donor, Supports Cellular Bioenergetics and Protects against Alzheimer's Disease by Preserving Mitochondrial Function in APP/PS1 Mice and Neurons

doi: 10.1155/2016/8360738

Figure Lengend Snippet: AP39 alleviated the brain atrophy of APP/PS1 mice as observed by brain magnetic resonance imaging (MRI). Brain MRI data were collected at different coronal and axial sections in 12-month-old mice receiving different treatments. Notes: panels (a), (c), and (e) show coronal sections, and panels (b), (d), and (f) show axial sections. The highlighted white regions indicate cerebrospinal fluid (CSF) in the ventricle, and the hippocampus is adjacent to the ventricle. Representative images from brain MRI depict slices of the T2-weighted morphologic images of 12-month-old WT mice treated with water (a)-(b) or AD mice treated with water (c)-(d) or AP39 (e)-(f). (g) ADC value determination of the mouse brains.

Article Snippet: AP39, a novel mitochondria-targeted H 2 S donor, was designed and synthesized by Medicilon Inc., as described previously [ , ].

Techniques: Magnetic Resonance Imaging

Journal: Oxidative Medicine and Cellular Longevity

Article Title: AP39, a Mitochondria-Targeted Hydrogen Sulfide Donor, Supports Cellular Bioenergetics and Protects against Alzheimer's Disease by Preserving Mitochondrial Function in APP/PS1 Mice and Neurons

doi: 10.1155/2016/8360738

Figure Lengend Snippet: AP39 reduced A β production and A β deposition in APP/PS1 mice. (a)-(b) After intraperitoneal injection of AP39 for 6 weeks, A β 40 and A β 42 levels in the mouse brains were decreased from 925 pg/mL and 532 pg/mL to 531 pg/mL and 365 pg/mL, respectively. (c) A β plaques were detected in the cortex and the hippocampus of the WT and APP/PS1 mouse brains based on 6E10 immunostaining. The statistical results show the area of the A β plaques. Original magnification: –40x; scale bars = 100 μ m.

Article Snippet: AP39, a novel mitochondria-targeted H 2 S donor, was designed and synthesized by Medicilon Inc., as described previously [ , ].

Techniques: Injection, Immunostaining

Journal: Experimental Animals

Article Title: AP39 ameliorates high fat diet-induced liver injury in young rats via alleviation of oxidative stress and mitochondrial impairment

doi: 10.1538/expanim.21-0056

Figure Lengend Snippet: Effect of AP39 on obesity. High-fat-diet (HFD)-induced non-alcoholic fatty liver disease (NAFLD) rats were intravenously injected with 0.05 or 0.1 mg/kg AP39 once a day for 7 weeks. (A) Schematic abstract of the experimental process. (B, C) Body weight and food intake of young rats was recorded once a week for 7 weeks. ** P< 0.01 vs. Control; ^ P< 0.05, ^^ P< 0.01 vs. HFD.

Article Snippet: To investigate the impact of AP39 on HFD rats, the rats in HFD+L-AP39 and HFD+H-AP39 groups were injected daily by tail vein with AP39 (APExBIO, Houston, TX, USA) at the dose of 0.05 mg/kg or 0.1 mg/kg, respectively [ ].

Techniques: Injection, Control

Journal: Experimental Animals

Article Title: AP39 ameliorates high fat diet-induced liver injury in young rats via alleviation of oxidative stress and mitochondrial impairment

doi: 10.1538/expanim.21-0056

Figure Lengend Snippet: Effect of AP39 on high-fat-diet (HFD)-induced non-alcoholic fatty liver disease (NAFLD). (A) At the end of the experiment, the liver tissues were harvested. Gross morphology of liver tissue was observed. (B) Liver weight index was calculated according to the formula liver wet weight / body weight × 100%. (C, D) The histological alternations of liver tissues were visualized with Oil red O staining (×200) and hematoxylin and eosin (H&E) staining (×200), and the representative photomicrographs were shown. (E) NAFLD activity score, a histological scoring system for NAFLD, was counted. ** P< 0.01 vs. Control; ^ P< 0.05, ^^ P< 0.01 vs. HFD.

Article Snippet: To investigate the impact of AP39 on HFD rats, the rats in HFD+L-AP39 and HFD+H-AP39 groups were injected daily by tail vein with AP39 (APExBIO, Houston, TX, USA) at the dose of 0.05 mg/kg or 0.1 mg/kg, respectively [ ].

Techniques: Staining, Activity Assay, Control

Journal: Experimental Animals

Article Title: AP39 ameliorates high fat diet-induced liver injury in young rats via alleviation of oxidative stress and mitochondrial impairment

doi: 10.1538/expanim.21-0056

Figure Lengend Snippet: Effect of AP39 on lipid profiles in serum of rats. (A–E) Relative levels of serum ALT, AST, TC, TG, LDL-C and HDL-C were determined using the corresponding kits. ALT, alanine transaminase; AST, aspartate transaminase; TC, total cholesterol; TG, total triglycerides; LDL-C, low-density lipoprotein cholesterol; HDL-C, high-density lipoprotein cholesterol. ** P< 0.01 vs. Control; ^ P< 0.05, ^^ P< 0.01 vs. HFD.

Article Snippet: To investigate the impact of AP39 on HFD rats, the rats in HFD+L-AP39 and HFD+H-AP39 groups were injected daily by tail vein with AP39 (APExBIO, Houston, TX, USA) at the dose of 0.05 mg/kg or 0.1 mg/kg, respectively [ ].

Techniques: Control

Journal: Experimental Animals

Article Title: AP39 ameliorates high fat diet-induced liver injury in young rats via alleviation of oxidative stress and mitochondrial impairment

doi: 10.1538/expanim.21-0056

Figure Lengend Snippet: Effect of AP39 on high-fat diet (HFD)-induced oxidative stress. (A) DHE staining (×400) was used for ROS detection in liver. (B–D) Levels of markers of oxidative stress, MDA, GSH and SOD, were measured with commercial kits. ROS, reactive oxygen species; MDA, malondialdehyde; GSH, glutathione; SOD, superoxide dismutase. ** P< 0.01 vs. Control; ^ P< 0.05, ^^ P< 0.01 vs. HFD.

Article Snippet: To investigate the impact of AP39 on HFD rats, the rats in HFD+L-AP39 and HFD+H-AP39 groups were injected daily by tail vein with AP39 (APExBIO, Houston, TX, USA) at the dose of 0.05 mg/kg or 0.1 mg/kg, respectively [ ].

Techniques: Staining, Control

Journal: Experimental Animals

Article Title: AP39 ameliorates high fat diet-induced liver injury in young rats via alleviation of oxidative stress and mitochondrial impairment

doi: 10.1538/expanim.21-0056

Figure Lengend Snippet: Effect of AP39 on H 2 S level and mitochondrial function. (A) Hepatic H 2 S level was detected using the H 2 S determination kit. (B) mtDNA copy number in the liver was measured by quantitative real-time polymerase chain reaction (RT-qPCR). (C) The degree of mitochondrial swelling was analyzed at 520 nm wavelength on a spectrophotometer. (D) Mitochondrial membrane potential (MMP) changes were detected using JC-1 probe. (E) Hypoxia-inducible factor-1α (HIF-1α) expression at mRNA and protein levels was determined by RT-qPCR and western blot, respectively. ** P< 0.01 vs. Control; ^ P< 0.05, ^^ P< 0.01 vs. high-fat diet (HFD).

Article Snippet: To investigate the impact of AP39 on HFD rats, the rats in HFD+L-AP39 and HFD+H-AP39 groups were injected daily by tail vein with AP39 (APExBIO, Houston, TX, USA) at the dose of 0.05 mg/kg or 0.1 mg/kg, respectively [ ].

Techniques: Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Spectrophotometry, Membrane, Expressing, Western Blot, Control