Nutr Res Pract.  2018 Jun;12(3):199-207. 10.4162/nrp.2018.12.3.199.

GABA-enriched fermented Laminaria japonica improves cognitive impairment and neuroplasticity in scopolamine- and ethanol-induced dementia model mice

• Reid SN
• Ryu JK
• Kim Y
• Jeon BH

• KMID: 2442293
• DOI: http://doi.org/10.4162/nrp.2018.12.3.199

Abstract

BACKGROUND/OBJECTIVES
Fermented Laminaria japonica (FL), a type sea tangle used as a functional food ingredient, has been reported to possess cognitive improving properties that may aid in the treatment of common neurodegenerative disorders, such as dementia.
MATERIALS/METHODS
We examined the effects of FL on scopolamine (Sco)- and ethanol (EtOH)-induced hippocampus-dependent memory impairment, using the Passive avoidance (PA) and Morris water maze (MWM) tests. To examine the underlying mechanisms associated with neuroprotective effects, we analyzed acetylcholine (ACh) and acetylcholinesterase (AChE) activity, brain tissue expression of muscarinic acetylcholine receptor (mAChR), cAMP response element binding protein (CREB) and extracellular signal-regulated kinases 1/2 (ERK1/2), and immunohistochemical analysis, in the hippocampus of mice, compared to current drug therapy intervention. Biochemical blood analysis was carried out to determine the effects of FL on alanine transaminase (ALT), aspartate transaminase (AST), and triglyceride (TG) and total cholesterol (TC) levels. 7 groups (n = 10) consisted of a control (CON), 3 Sco-induced dementia and 3 EtOH-induced dementia groups, with both dementia group types containing an untreated group (Sco and EtOH); a positive control, orally administered donepezil (Dpz) (4mg/kg) (Sco + Dpz and EtOH + Dpz); and an FL (50 mg/kg) treatment group (Sco + FL50 and EtOH + FL50), orally administered over the 4-week experimental period.
RESULTS
FL50 significantly reduced EtOH-induced increase in AST and ALT levels. FL50 treatment reduced EtOH-impaired step-through latency time in the PA test, and Sco- and EtOH-induced dementia escape latency times in the MWM test. Moreover, anticholinergic effects of Sco and EtOH on the brain were reversed by FL50, through the attenuation of AChE activity and elevation of ACh concentration. FL50 elevated ERK1/2 protein expression and increased p-CREB (ser133) in hippocampus brain tissue, according to Western blot and immunohistochemistry analysis, respectively.
CONCLUSION
Overall, these results suggest that FL may be considered an efficacious intervention for Sco- and EtOH-induced dementia, in terms of reversing cognitive impairment and neuroplastic dysfunction.

Keyword

Fermented laminaria; cognitive dysfunction; acetylcholine; cyclic AMP response element; muscarinic receptors

MeSH Terms

Acetylcholine
Acetylcholinesterase
Alanine Transaminase
Animals
Aspartate Aminotransferases
Blotting, Western
Brain
Cholesterol
Cognition Disorders*
Cyclic AMP Response Element-Binding Protein
Dementia*
Drug Therapy
Ethanol
Extracellular Signal-Regulated MAP Kinases
Functional Food
Hippocampus
Immunohistochemistry
Laminaria*
Memory
Mice*
Neurodegenerative Diseases
Neuronal Plasticity*
Neuroprotective Agents
Receptors, Muscarinic
Scopolamine Hydrobromide
Triglycerides
United Nations
Water
Acetylcholine
Acetylcholinesterase
Alanine Transaminase
Aspartate Aminotransferases
Cholesterol
Cyclic AMP Response Element-Binding Protein
Ethanol
Extracellular Signal-Regulated MAP Kinases
Neuroprotective Agents
Receptors, Muscarinic
Scopolamine Hydrobromide
Water

Figure

• 

  Fig. 1 Experimental design FL50, Fermented Laminaria japonica 50; ALT, alanine transaminase; AST, aspartate transaminase;TG, triglyceride; TC, total cholesterol; ACh, acetylcholine; AChE, acetylcholinesterase.

• 

  Fig. 2 Effects of FL50 on ALT, AST, TG and TC of Sco- and EtOH-induced memory impaired mice 1)Values expressed as mean ± SD. 2) †P < 0.05 vs. EtOH. ALT, alanine transaminase; AST, aspartate transaminase; CON, control; Sco, scopolamine; EtOH, ethanol; Dpz, donepezil; FL50, Fermented Laminaria japonica 50.

• 

  Fig. 3 Morris water maze test: Effects of FL50 on hippocampus-dependent spatial learning ability in Sco- and EtOH-induced memory impaired mice 1)Values are expressed as mean ± SD. 2) *P < 0.05 vs. Sco, †P < 0.05 vs. EtOH. CON, control; Sco, scopolamine; EtOH, ethanol; Dpz, donepezil; FL50, Fermented Laminaria japonica 50.

• 

  Fig. 4 Passive avoidance test: Effects of FL50 on short-term working memory ability in Sco- and EtOH-induced memory impaired mice 1) **P < 0.001 vs. Sco, ††P < 0.001 vs. EtOH. CON, control; Sco, scopolamine; EtOH, ethanol; Dpz, donepezil; FL50, Fermented Laminaria japonica 50.

• 

  Fig. 5 Effects of FL50 on ACh concentration and AChE activity in Sco- and EtOH-induced memory impaired mice 1)Values expressed as mean ± SD. 2) *P < 0.05 vs. Sco, **P < 0.001 vs. Sco, †P < 0.05 vs. EtOH, ††P < 0.001 vs. EtOH. CON, control; Sco, scopolamine; EtOH, ethanol; Dpz, donepezil; FL50, Fermented Laminaria japonica 50.

• 

  Fig. 6 Effects of FL50 on hippocampus protein expression in Sco- and EtOH-induced memory impaired mice: (A) Western blot analysis showing mAChR, p-ERK1/2, ERK1/2, p-CREB (Ser133), CREB and GADPH protein expression (B) p-ERK1/2/ERK1/2 ratio (C) mAChR/GADPH ratio and (D) p-CREB (Ser133)/CREB ratio 1)Values are expressed as mean ± SD. 2) *P < 0.05 vs. Sco, †P < 0.05 vs. EtOH. CON, control; Sco, scopolamine; EtOH, ethanol; Dpz, donepezil; FL50, Fermented Laminaria japonica 50.

• 

  Fig. 7 Effects of FL50 on expression of p-CREB (Ser133) in the hippocampus of Sco- and EtOH-induced dementia mice (Immunohistochemistry original magnification, × 100). CON, control; Sco, scopolamine; EtOH, ethanol; Dpz, donepezil; FL50, Fermented Laminaria japonica 50.

• 

  Fig. 8 Regulation of mAChR, ERK1/2 and CREB in the brain BDNF, brain-derived neurotrophic factor; TrkB, Tropomyosin receptor kinase B; PI3K/Akt, phosphatidylinositol 3-kinase/Akt; MEK, mitogen-activated ERK kinas; PKC, protein kinase C; PLC, phospholipase C; DAG, diacylgycerol; IP3, inositol 1,4,5-trisphosphate; MAPK, mitogen-activated protein kinase; ERK, extracellular signal regulated kinase; CaMK, Calcium/calmodulin-dependent protein kinase; CDK2/CDK4, cyclin dependent kinases 2/4.

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