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

Yun Jeong Kim; Chung Soo Lee

doi:10.4196/kjpp.2008.12.2.65

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

Original Article

Korean J Physiol Pharmacol 2008;12(2):65-71.

Published online: 17 April 2008

DOI: https://doi.org/10.4196/kjpp.2008.12.2.65

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

Yun Jeong Kim, Chung Soo Lee

Department of Pharmacology, College of Medicine, Chung-Ang University, Seoul 156-756, Korea

Corresponding to: Chung Soo Lee, Department of Pharmacology, College of Medicine, Chung-Ang University, 211, Heukseok-dong, Dongjak-gu, Seoul 156-756, Korea. (Tel) 82-2-820-5659, (Fax) 82-2-815-3856, (E-mail) leecs@cau.ac.kr

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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.

Keywords: Glycyrrhizin, MPTP, MPP⁺, Brain tissue damage, Cell death, Inhibitory 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.

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.

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).

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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