Journal List > Korean J Physiol Pharmacol > v.12(2) > 1025580

Kim and Lee: Glycyrrhizin Attenuates MPTP Neurotoxicity in Mouse and MPP+- Induced Cell Death in PC12 Cells

Abstract

The present study examined the inhibitory effect of licorice compounds glycyrrhizin and a metabolite 18β-glycyrrhetinic acid on the neurotoxicity of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in the mouse and on the 1-methyl-4-phenylpyridinium (MPP+)-induced cell death in differentiated PC12 cells. MPTP treatment increased the activities of total superoxide dismutase, catalase and glutathione peroxidase and the levels of malondialdehyde and carbonyls in the brain compared to control mouse brain. Co-administration of glycyrrhizin (16.8 mg/kg) attenuated the MPTP effect on the enzyme activities and formation of tissue peroxidation products. In vitro assay, licorice compounds attenuated the MPP+-induced cell death and caspase-3 activation in PC12 cells. Glycyrrhizin up to 100μM significantly attenuated the toxicity of MPP+. Meanwhile, 18β-glycyrrhetinic acid showed a maximum inhibitory effect at 10μM; beyond this concentration the inhibitory effect declined. Glycyrrhizin and 18β-glycyrrhetinic acid attenuated the hydrogen peroxide- or nitrogen species-induced cell death. Results from this study indicate that glycyrrhizin may attenuate brain tissue damage in mice treated with MPTP through inhibitory effect on oxidative tissue damage. Glycyrrhizin and 18β-glycyrrhetinic acid may reduce the MPP+ toxicity in PC12 cells by suppressing caspase-3 activation. The effect seems to be ascribed to the antioxidant effect.

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Fig. 1.
Effect of licorice compounds on MPP+-induced cell death. PC12 cells were pre-treated with licorice compounds (1~100 μM GL in A or 1-25 μM GA in B) for 20 min, exposed to 500 μM MPP+ for 24 h and cell viability was determined. Data represent means ± SEM (n=6). +p<0.05 compared to control (percentage of control); and ∗p<0.05 compared to MPP+ alone.
kjpp-12-65f1.tif
Fig. 2.
Effect of licorice compounds on MPP+-induced activation of caspase-3. PC12 cells were treated with 500 μM MPP+ in the presence of licorice compounds (10~50 μM) or scavengers [1 mM N-acetylcysteine (NAC), 30 μM trolox or 30 μM carboxy-PTIO (PTIO)] for 24 h. Data are expressed as units for caspase-3 activity and represent means ± SEM (n=6). +p<0.05 compared to control; and ∗p<0.05 compared to MPP+ alone.
kjpp-12-65f2.tif
Fig. 3.
Effect of licorice compounds on cell death due to hydrogen peroxide or 3-morpholinosydnonime. PC12 cells were pre-treated with licorice compounds [1~50 μM GL (A) or 1~10 μM GA (B)] for 15 min, exposed to 200 μM hydrogen peroxide for 4 h or 750 μM 3-morpholinosydnonime for 24 h, and cell viability was determined. Data represent means ± SEM (n=6). +p<0.05 compared to control; and ∗p<0.05 compared to hydrogen peroxide (HP) or 3-morpholinosydnonime (SIN).
kjpp-12-65f3.tif
Table 1.
Effect of GL on increased antioxidant enzyme activities in the brains of mice treated with MPTP
  Control MPTP MPTP+GL GL
BDM        
 SOD 23.16±0.73 39.32±1.19 28.86±0.96 24.58±0.81
 Catalase 0.603±0.024 0.926±0.025 0.723±0.023 0.603±0.022
 GSH peroxidase 3.999±0.111 6.269±0.173 5.003±0.172 4.276±0.136
Cortex        
 SOD 7.98±0.22 9.95±0.31 8.94±0.22 7.78±0.23
 Catalase 0.384±0.032 0.520±0.025 0.428±0.034 0.406±0.027
 GSH peroxidase 3.698±0.129 4.984±0.145 4.120±0.129 3.778±0.146
Cerebellum        
 SOD 11.84±0.47 17.23±0.67 13.66±0.75 11.43±0.45
 Catalase 0.857±0.039 1.123±0.040 0.920±0.043 0.841±0.037
 GSH peroxidase 5.145±0.256 7.014±0.117 5.908±0.266 5.366±0.198

Mice were treated with MPTP (30 mg/kg) and GL (16.8 mg/kg) for 5 days. Enzyme activities in tissue homogenates of BDM, cortex and cerebellum were measured as described in Methods. Data represent units/mg protein and means ± SEM (n=8).

p<0.05 compared to control

p<0.05 compared to MPTP alone.

Table 2.
Effect of GL on formation of MDA and carbonyls in the brains of mice treated with MPTP
  Control MPTP MPTP+GL GL
BDM        
 MDA 0.268±0.010 0.462±0.017 0.340±0.014 0.282±0.012
 Carbonyls 1.835±0.059 2.918±0.102 2.248±0.117 1.907±0.079
Cortex        
 MDA 0.292±0.012 0.384±0.019 0.328±0.013 0.285±0.014
 Carbonyls 1.875±0.085 2.494±0.133 2.120±0.129 1.973±0.089
Cerebellum        
 MDA 0.306±0.012 0.409±0.016 0.358±0.014 0.273±0.013
 Carbonyls 1.971±0.085 2.705±0.090 2.238±0.103 1.890±0.082

Mice were treated with MPTP (30 mg/kg) and GL (16.8 mg/kg) for 5 days. Amounts of MDA and carbonyls in tissue homogenates of BDM, cortex and cerebellum were quantified as described in Methods. Data represent nmol/mg protein and means ± SEM (n=8).

p<0.05 compared to control

p<0.05 compared to MPTP alone.

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