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TRO40303, a mitochondrial-targeted cytoprotective compound, provides protection in hepatitis models.

Schaller S, Michaud M, Latyszenok V, Robert F, Hocine M, Arnoux T, Gabriac M, Codoul H, Bourhane A, de Bellefois IC, Afxantidis J, Pruss RM - Pharmacol Res Perspect (2015)

Bottom Line: When higher, lethal doses of Jo2 were administered, TRO40303 (10 and 30 mg/kg) significantly reduced mortality by 65-90% when administered intraperitoneally (i.p.) 1 h before Jo2 injection, a time when TRO40303 plasma concentrations reached their peak.TRO40303 (30 mg/kg, i.p.) was also able to reduce mortality by 30-50% when administered 1 h postlethal Jo2 intoxication.These results suggest that TRO40303 could be a promising new therapy for the treatment or prevention of hepatitis.

View Article: PubMed Central - PubMed

Affiliation: Trophos S. A., Luminy Biotech Entreprise Marseille, France.

ABSTRACT
TRO40303 is cytoprotective compound that was shown to reduce infarct size in preclinical models of myocardial infarction. It targets mitochondria, delays mitochondrial permeability transition pore (mPTP) opening and reduces oxidative stress in cardiomyocytes submitted to ischemia/reperfusion in vitro. Because the involvement of the mitochondria and the mPTP has been demonstrated in chronic as well as acute hepatitis, we investigated the potential of TRO40303 to prevent hepatocyte injury. A first set of in vitro studies showed that TRO40303 (from 0.3 to 3 μmol/L) protected HepG2 cells and primary mouse embryonic hepatocytes (PMEH) from palmitate intoxication, a model mimicking steatohepatitis. In PMEH, TRO40303 provided similar protection against cell death due to Jo2 anti-Fas antibody intoxication. Further studies were then preformed in a mouse model of Fas-induced fulminant hepatitis induced by injecting Jo2 anti-Fas antibody. When mice received a sublethal dose of Jo2 at 125 μg/kg, TRO40303 pretreatment prevented liver enzyme elevation in plasma in parallel with a decrease in cytochrome C release from mitochondria and caspase 3 and 7 activation in hepatic tissue. When higher, lethal doses of Jo2 were administered, TRO40303 (10 and 30 mg/kg) significantly reduced mortality by 65-90% when administered intraperitoneally (i.p.) 1 h before Jo2 injection, a time when TRO40303 plasma concentrations reached their peak. TRO40303 (30 mg/kg, i.p.) was also able to reduce mortality by 30-50% when administered 1 h postlethal Jo2 intoxication. These results suggest that TRO40303 could be a promising new therapy for the treatment or prevention of hepatitis.

No MeSH data available.


Related in: MedlinePlus

TRO40303 provides protection in hepatocyte intoxication models in vitro. Primary mouse embryonic hepatocytes (PMEH) were isolated as described in material and methods section. The cells (PMEH or HepG2) were pretreated for 1 h with compounds at a final constant DMSO concentration (0.1%) (n = 8 for all conditions), then intoxicated in the presence of compounds as follows. For palmitate intoxication, PMEH were intoxicated for 24 h at 0.2 mmol/L and viability was assessed using calcein staining (A), whereas HepG2 were subjected to 48 h 0.2 mmol/L palmitate intoxication for LDH cell death assessment (B). For anti-Fas antibody intoxication, PMEH were intoxicated for 24 h with 100 μg/mL Jo2 and 2 μg/mL CHX and viability was assessed using calcein staining (C). Results are presented as mean ± SEM and statistical analysis was performed by one-way analysis of variance followed by Dunnett’s posttest compared to the toxic control (**P < 0.01, ***P < 0.001). Survival of control, nontreated cells (NT), except for DMSO, is shown for comparison. Data shown are for a representative example of three independent experiments performed using three different cell culture preparation. CHX, cycloheximide; DMSO, dimethyl sulfoxide.
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fig01: TRO40303 provides protection in hepatocyte intoxication models in vitro. Primary mouse embryonic hepatocytes (PMEH) were isolated as described in material and methods section. The cells (PMEH or HepG2) were pretreated for 1 h with compounds at a final constant DMSO concentration (0.1%) (n = 8 for all conditions), then intoxicated in the presence of compounds as follows. For palmitate intoxication, PMEH were intoxicated for 24 h at 0.2 mmol/L and viability was assessed using calcein staining (A), whereas HepG2 were subjected to 48 h 0.2 mmol/L palmitate intoxication for LDH cell death assessment (B). For anti-Fas antibody intoxication, PMEH were intoxicated for 24 h with 100 μg/mL Jo2 and 2 μg/mL CHX and viability was assessed using calcein staining (C). Results are presented as mean ± SEM and statistical analysis was performed by one-way analysis of variance followed by Dunnett’s posttest compared to the toxic control (**P < 0.01, ***P < 0.001). Survival of control, nontreated cells (NT), except for DMSO, is shown for comparison. Data shown are for a representative example of three independent experiments performed using three different cell culture preparation. CHX, cycloheximide; DMSO, dimethyl sulfoxide.

Mentions: Palmitate intoxication of PMEH or human HepG2 cells has been proposed as a model of hepatic steatosis (Gomez-Lechon et al. 2007; Srivastava and Chan 2007). Both cell models were used to evaluate the hepatoprotective effects of TRO40303 in vitro against palmitate intoxication in a first set of experiments. The effect of palmitate was analyzed by looking at PMEH cell survival using the calcein assay using the Plate-Runner® in the Global fluorescent mode, whereas HepG2 cell death was analyzed using LDH release assessment. TRO40303 at doses from 0.3 to 3 μmol/L protected both PMEH and human HepG2 cells intoxicated with palmitate (Fig.1A and B).


TRO40303, a mitochondrial-targeted cytoprotective compound, provides protection in hepatitis models.

Schaller S, Michaud M, Latyszenok V, Robert F, Hocine M, Arnoux T, Gabriac M, Codoul H, Bourhane A, de Bellefois IC, Afxantidis J, Pruss RM - Pharmacol Res Perspect (2015)

TRO40303 provides protection in hepatocyte intoxication models in vitro. Primary mouse embryonic hepatocytes (PMEH) were isolated as described in material and methods section. The cells (PMEH or HepG2) were pretreated for 1 h with compounds at a final constant DMSO concentration (0.1%) (n = 8 for all conditions), then intoxicated in the presence of compounds as follows. For palmitate intoxication, PMEH were intoxicated for 24 h at 0.2 mmol/L and viability was assessed using calcein staining (A), whereas HepG2 were subjected to 48 h 0.2 mmol/L palmitate intoxication for LDH cell death assessment (B). For anti-Fas antibody intoxication, PMEH were intoxicated for 24 h with 100 μg/mL Jo2 and 2 μg/mL CHX and viability was assessed using calcein staining (C). Results are presented as mean ± SEM and statistical analysis was performed by one-way analysis of variance followed by Dunnett’s posttest compared to the toxic control (**P < 0.01, ***P < 0.001). Survival of control, nontreated cells (NT), except for DMSO, is shown for comparison. Data shown are for a representative example of three independent experiments performed using three different cell culture preparation. CHX, cycloheximide; DMSO, dimethyl sulfoxide.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4492760&req=5

fig01: TRO40303 provides protection in hepatocyte intoxication models in vitro. Primary mouse embryonic hepatocytes (PMEH) were isolated as described in material and methods section. The cells (PMEH or HepG2) were pretreated for 1 h with compounds at a final constant DMSO concentration (0.1%) (n = 8 for all conditions), then intoxicated in the presence of compounds as follows. For palmitate intoxication, PMEH were intoxicated for 24 h at 0.2 mmol/L and viability was assessed using calcein staining (A), whereas HepG2 were subjected to 48 h 0.2 mmol/L palmitate intoxication for LDH cell death assessment (B). For anti-Fas antibody intoxication, PMEH were intoxicated for 24 h with 100 μg/mL Jo2 and 2 μg/mL CHX and viability was assessed using calcein staining (C). Results are presented as mean ± SEM and statistical analysis was performed by one-way analysis of variance followed by Dunnett’s posttest compared to the toxic control (**P < 0.01, ***P < 0.001). Survival of control, nontreated cells (NT), except for DMSO, is shown for comparison. Data shown are for a representative example of three independent experiments performed using three different cell culture preparation. CHX, cycloheximide; DMSO, dimethyl sulfoxide.
Mentions: Palmitate intoxication of PMEH or human HepG2 cells has been proposed as a model of hepatic steatosis (Gomez-Lechon et al. 2007; Srivastava and Chan 2007). Both cell models were used to evaluate the hepatoprotective effects of TRO40303 in vitro against palmitate intoxication in a first set of experiments. The effect of palmitate was analyzed by looking at PMEH cell survival using the calcein assay using the Plate-Runner® in the Global fluorescent mode, whereas HepG2 cell death was analyzed using LDH release assessment. TRO40303 at doses from 0.3 to 3 μmol/L protected both PMEH and human HepG2 cells intoxicated with palmitate (Fig.1A and B).

Bottom Line: When higher, lethal doses of Jo2 were administered, TRO40303 (10 and 30 mg/kg) significantly reduced mortality by 65-90% when administered intraperitoneally (i.p.) 1 h before Jo2 injection, a time when TRO40303 plasma concentrations reached their peak.TRO40303 (30 mg/kg, i.p.) was also able to reduce mortality by 30-50% when administered 1 h postlethal Jo2 intoxication.These results suggest that TRO40303 could be a promising new therapy for the treatment or prevention of hepatitis.

View Article: PubMed Central - PubMed

Affiliation: Trophos S. A., Luminy Biotech Entreprise Marseille, France.

ABSTRACT
TRO40303 is cytoprotective compound that was shown to reduce infarct size in preclinical models of myocardial infarction. It targets mitochondria, delays mitochondrial permeability transition pore (mPTP) opening and reduces oxidative stress in cardiomyocytes submitted to ischemia/reperfusion in vitro. Because the involvement of the mitochondria and the mPTP has been demonstrated in chronic as well as acute hepatitis, we investigated the potential of TRO40303 to prevent hepatocyte injury. A first set of in vitro studies showed that TRO40303 (from 0.3 to 3 μmol/L) protected HepG2 cells and primary mouse embryonic hepatocytes (PMEH) from palmitate intoxication, a model mimicking steatohepatitis. In PMEH, TRO40303 provided similar protection against cell death due to Jo2 anti-Fas antibody intoxication. Further studies were then preformed in a mouse model of Fas-induced fulminant hepatitis induced by injecting Jo2 anti-Fas antibody. When mice received a sublethal dose of Jo2 at 125 μg/kg, TRO40303 pretreatment prevented liver enzyme elevation in plasma in parallel with a decrease in cytochrome C release from mitochondria and caspase 3 and 7 activation in hepatic tissue. When higher, lethal doses of Jo2 were administered, TRO40303 (10 and 30 mg/kg) significantly reduced mortality by 65-90% when administered intraperitoneally (i.p.) 1 h before Jo2 injection, a time when TRO40303 plasma concentrations reached their peak. TRO40303 (30 mg/kg, i.p.) was also able to reduce mortality by 30-50% when administered 1 h postlethal Jo2 intoxication. These results suggest that TRO40303 could be a promising new therapy for the treatment or prevention of hepatitis.

No MeSH data available.


Related in: MedlinePlus