Lab Anim Res.  2016 Mar;32(1):24-33. 10.5625/lar.2016.32.1.24.

Role of mitogen-activated protein kinases and nuclear factor-kappa B in 1,3-dichloro-2-propanol-induced hepatic injury

• Lee IC
• Lee SM
• Ko JW
• Park SH
• Shin IS
• Moon C
• Kim SH
• Kim JC

• KMID: 2312151
• DOI: http://doi.org/10.5625/lar.2016.32.1.24

Abstract

In this study, the potential hepatotoxicity of 1,3-dichloro-2-propanol and its hepatotoxic mechanisms in rats was investigated. The test chemical was administered orally to male rats at 0, 27.5, 55, and 110 mg/kg body weight. 1,3-Dichloro-2-propanol administration caused acute hepatotoxicity, as evidenced by an increase in serum aminotransferases, total cholesterol, and total bilirubin levels and a decrease in serum glucose concentration in a dose-dependent manner with corresponding histopathological changes in the hepatic tissues. The significant increase in malondialdehyde content and the significant decrease in glutathione content and antioxidant enzyme activities indicated that 1,3-dichloro-2-propanol-induced hepatic damage was mediated through oxidative stress, which caused a dose-dependent increase of hepatocellular apoptotic changes in the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay and immunohistochemical analysis for caspase-3. The phosphorylation of mitogen-activated protein kinases caused by 1,3-dichloro-2-propanol possibly involved in hepatocellular apoptotic changes in rat liver. Furthermore, 1,3-dichloro-2-propanol induced an inflammatory response through activation of nuclear factor-kappa B signaling that coincided with the induction of pro-inflammatory mediators or cytokines in a dose-dependent manner. Taken together, these results demonstrate that hepatotoxicity may be related to oxidative stress-mediated activation of mitogen-activated protein kinases and nuclear factor-kappa B-mediated inflammatory response.

Keyword

1,3-dichloro-2-propanol; hepatotoxicity; MAPKs; NF-κB

MeSH Terms

Animals
Bilirubin
Blood Glucose
Body Weight
Caspase 3
Cholesterol
Cytokines
Glutathione
Humans
Liver
Male
Malondialdehyde
Mitogen-Activated Protein Kinases*
Oxidative Stress
Phosphorylation
Rats
Transaminases
Bilirubin
Caspase 3
Cholesterol
Cytokines
Glutathione
Malondialdehyde
Mitogen-Activated Protein Kinases
Transaminases

Figure

• 

  Figure 1 Effects of 1,3-DCP on hepatic histopathology. Representative photographs of liver sections of (A) control showing normal appearance and rats treated with 1,3-DCP at doses of (B) 27.5, (C) 55, and (D) 110 mg/kg showing various histopathological alterations characterized by degeneration/necrosis of hepatocytes around the central vein region (open arrowheads), vacuolation (closed arrows), inflammatory cell infiltration (open arrows), hemorrhage (asterisks), and sinusoidal dilation (closed arrowheads). Hematoxylin and eosin stain. Bar=50 µm (×200). 1,3-DCP: 1,3-dichloro-2-propanol.

• 

  Figure 2 Effects of 1,3-DCP on hepatic cellular apoptosis. Representative photographs of TUNEL assay (A-D) and immunohistochemical analysis of caspase-3 (E-H) performed on liver sections. (A, E) Liver of control rats showed scant positive cells. The rats treated with 1,3-DCP at doses of (B, F) 27.5, (C, G) 55, and (D, H) 110 mg/kg showed a dose-dependent increase in TUNEL- and caspase-3-positive cells. Bar=50 µm (×200). 1,3-DCP: 1,3-dichloro-2-propanol; and TUNEL: terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling.

• 

  Figure 3 Effects of 1,3-DCP on MAPKs-dependent pathways. Immunoblotting analysis of (A) MAPKs-related Erk, JNK, p38 MAPK and its phosphorylated form protein levels in male rats treated with 1,3-DCP (loading control: β-actin). The bar graphs show relative (B) p-Erk/total Erk (t-Erk), (C) p-JNK/t-JNK, and (D) p-p38 MAPK/t-p38 MAPK ratios in hepatic tissues for 1,3-DCP-treated rats. Values are presented as mean±SD (n=6). *P<0.05 versus control group, **P<0.01 versus control group. 1,3-DCP: 1,3-dichloro-2-propanol; MAPKs: mitogen-activated protein kinases; Erk: p44/42 MAPK; and JNK: c-Jun N-terminal kinase.

• 

  Figure 4 Effects of 1,3-DCP on NF-κB expression. (A) Immunoblotting of hepatic nuclear NF-κB expression in the male rats treated with 1,3-DCP (nuclear loading control: Lamin B1). (B) The bar graphs show relative nuclear NF-κB protein levels in hepatic tissues for 1,3-DCP treated rats. Values are presented as mean±SD (n=6). **P<0.01 versus control group. 1,3-DCP: 1,3-dichloro-2-propanol; and NF-κB: nuclear factor-kappa B.

• 

  Figure 5 Effects of 1,3-DCP on NF-κB-related inflammatory mediators and cytokines. (A) Immunoblotting analysis of TNF-α, Cox-2, and iNOS expression and (B) its relative protein levels in the male rats treated with 1,3-DCP (loading control: β-actin). The bar graphs show relative mRNA levels of NF-κB-related inflammatory mediator or cytokine, (C) iNOS, (D) Cox-2, (E) TNF-α, (F) IL-1β, and (G) IL-6 mRNA levels for 1,3-DCP-treated rat (loading control: GAPDH). Values are presented as mean±SD (n=6). **P<0.01 versus control group. 1,3-DCP: 1,3-dichloro-2-propanol; NF-κB: nuclear factor-kappa B; TNF-α: tumor necrosis factor-alpha; Cox-2: cyclooxygenase-2; iNOS: inducible nitric oxide synthase; IL-1β: interleukin-1β; IL-6: interleukin-6; and GAPDH: glyceraldehydes-3-phosphate dehydrogenase.

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