Korean J Physiol Pharmacol.  2015 Jul;19(4):365-372. 10.4196/kjpp.2015.19.4.365.

Immunotoxicological Effects of Aripiprazole: In vivo and In vitro Studies

• Baek KS
• Ahn S
• Lee J
• Kim JH
• Kim HG
• Kim E
• Kim JH
• Sung NY
• Yang S
• Kim MS
• Hong S
• Kim JH
• Cho JY

• KMID: 2285580
• DOI: http://doi.org/10.4196/kjpp.2015.19.4.365

Abstract

Aripiprazole (ARI) is a commonly prescribed medication used to treat schizophrenia and bipolar disorder. To date, there have been no studies regarding the molecular pathological and immunotoxicological profiling of aripiprazole. Thus, in the present study, we prepared two different formulas of aripiprazole [Free base crystal of aripiprazole (ARPGCB) and cocrystal of aripiprazole (GCB3004)], and explored their effects on the patterns of survival and apoptosis-regulatory proteins under acute toxicity and cytotoxicity test conditions. Furthermore, we also evaluated the modulatory activity of the different formulations on the immunological responses in macrophages primed by various stimulators such as lipopolysaccharide (LPS), pam3CSK, and poly(I:C) via toll-like receptor 4 (TLR4), TLR2, and TLR3 pathways, respectively. In liver, both ARPGCB and GCB3004 produced similar toxicity profiles. In particular, these two formulas exhibited similar phospho-protein profiling of p65/nuclear factor (NF)-kappaB, c-Jun/activator protein (AP)-1, ERK, JNK, p38, caspase 3, and bcl-2 in brain. In contrast, the patterns of these phospho-proteins were variable in other tissues. Moreover, these two formulas did not exhibit any cytotoxicity in C6 glioma cells. Finally, the two formulations at available in vivo concentrations did not block nitric oxide (NO) production from activated macrophage-like RAW264.7 cells stimulated with LPS, pam3CSK, or poly(I:C), nor did they alter the morphological changes of the activated macrophages. Taken together, our present work, as a comparative study of two different formulas of aripiprazole, suggests that these two formulas can be used to achieve similar functional activation of brain proteins related to cell survival and apoptosis and immunotoxicological activities of macrophages.

Keyword

Acute toxicity; Aripiprazole; Cocrystal; Immunotoxicology; Survival proteins

MeSH Terms

Aripiprazole
Apoptosis
Bipolar Disorder
Brain
Caspase 3
Cell Survival
Glioma
Liver
Macrophages
Nitric Oxide
Schizophrenia
Toll-Like Receptor 4
Caspase 3
Nitric Oxide
Toll-Like Receptor 4

Figure

• 

  Fig. 1 Acute toxicity of ARPGCB or GCB3004 in mice. (A) The levels of serum parameters (AST and ALT) from mice orally administered ARPGCB or GCB3004 (each 500 mg/kg). (B) Organs from mice orally administered with ARPGCB or GCB3004 (each 500 mg/kg). Data represent the mean±SD of an experiment performed with five mice.

• 

  Fig. 2 Effect of ARPGCB or GCB3004 on the level of cell death-related molecules in organs. (A, B, and C) The levels of cell death-related proteins such as (p65/NF-κB, c-Jun/AP-1, ERK/MAPK, JNK/MAPK, p38/MAPK, caspase 3, bcl-2, and β-actin) were identified by immunoblotting from tissue lysates prepared from mice orally administered with ARPGCB or GCB3004 (each of 500 mg/kg). The results shown are representative of three independent experiments.

• 

  Fig. 3 The effect of ARPGCB or GCB3004 on the viability of C6 glioma cells. (A) C6 glioma cells (2×106 cells/ml) were incubated with ARPGCB or GCB3004 for 24 h. Cell viability was evaluated by conventional MTT assay. (B) Images of the cells in culture at 12 h were obtained with an inverted phase contrast microscope that was interfaced with a video camera using Image J software.

• 

  Fig. 4 The effect of ARPGCB or GCB3004 on the immunological responses of RAW264.7 cells. (A) RAW264.7 cells (2×106 cells/ml) were incubated with ARPGCB or GCB3004 for 24 h. Cell viability was evaluated by conventional MTT assay. (B~D) Regulatory activity of ARPGCB or GCB3004 on the production of NO from activated macrophages stimulated by LPS (1 µg/ml), poly(I:C) (200 µg/ml), or Zymosan A (Zymo A, 400 µg/ml) was determined by Griess assay. (E) Images of cells in culture at the indicated time points were obtained with an inverted phase contrast microscope that was interfaced with a video camera using Image J software. **p<0.01 compared to each control group.

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