Endocrinol Metab.  2014 Sep;29(3):336-348. 10.3803/EnM.2014.29.3.336.

Growth Hormone-Releaser Diet Attenuates Cognitive Dysfunction in Klotho Mutant Mice via Insulin-Like Growth Factor-1 Receptor Activation in a Genetic Aging Model

• Park SJ
• Chung YH
• Lee JH
• Dang DK
• Nam Y
• Jeong JH
• Kim YS
• Nabeshima
• Shin EJ
• Kim HC

• KMID: 2169471
• DOI: http://doi.org/10.3803/EnM.2014.29.3.336

Abstract

BACKGROUND
It has been recognized that a defect in klotho gene expression accelerates the degeneration of multiple age-sensitive traits. Accumulating evidence indicates that aging is associated with declines in cognitive function and the activity of growth hormone (GH)/insulin-like growth factor-1 (IGF-1).
METHODS
In this study, we examined whether a GH-releaser diet could be effective in protecting against cognitive impairment in klotho mutant mice.
RESULTS
The GH-releaser diet significantly induced the expression of IGF-1 and IGF-1 receptors in the hippocampus of klotho mutant mice. Klotho mutant mice showed significant memory impairments as compared with wild-type mice. In addition, the klotho mutation significantly decreased the expression of cell survival/antiapoptotic factors, including phospho-Akt (p-Akt)/phospho-glycogen synthase kinase3beta (p-GSK3beta), phospho-extracellular signal-related kinase (p-ERK), and Bcl-2, but significantly increased those of cell death/proapoptotic factors, such as phospho-c-jun N-terminal kinase (p-JNK), Bax, and cleaved caspase-3 in the hippocampus. Treatment with GH-releaser diet significantly attenuated both decreases in the expression of cell survival/antiapoptotic factors and increases in the expression of cell death/proapoptotic factors in the hippocampus of klotho mutant mice. In addition, klotho mutation-induced oxidative stress was significantly attenuated by the GH-releaser diet. Consequently, a GH-releaser diet significantly improved memory function in the klotho mutant mice. GH-releaser diet-mediated actions were significantly reversed by JB-1, an IGF-1 receptor antagonist.
CONCLUSION
The results suggest that a GH-releaser diet attenuates oxidative stress, proapoptotic changes and consequent dysfunction in klotho mutant mice by promoting IGF-1 expression and IGF-1 receptor activation.

Keyword

klotho gene; Cognitive dysfunction; GH-releaser diet; Receptor, IGF type 1

MeSH Terms

Aging*
Animals
Caspase 3
Diet*
Gene Expression
Growth Hormone
Hippocampus
Insulin-Like Growth Factor I
Memory
Mice*
Oxidative Stress
Phosphotransferases
Receptor, IGF Type 1
Caspase 3
Growth Hormone
Insulin-Like Growth Factor I
Phosphotransferases
Receptor, IGF Type 1

Figure

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  Fig. 1 Experimental schedule for evaluating the effect of a growth hormone (GH)-releaser diet on the cognitive dysfunction in klotho mutant mice. Mice received a GH-releaser diet or a vehicle diet during the entire experimental period. JB-1, an insulin-like growth factor-1 (IGF-1) receptor antagonist, was infused (1 µg/5 µL/5 min, once daily for 25 days) into the lateral ventricle. Twenty days after the GH releaser diet began, a novel object recognition test (NORT) and a conditioned fear learning test (CFL) were performed. Mice were sacrificed for Western blot, reverse transcription-polymerase chain reaction (RT-PCR), or neurochemical analyses at 15 minutes after the CFL.

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  Fig. 2 Effect of a growth hormone (GH)-releaser diet on the mRNA expression of insulin-like growth factor-1 (IGF-1) and IGF-1 receptor in the hippocampus of klotho mutant mice. Each value is the mean±SEM of six animals. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; aCSF, artificial cerebrospinal fluid; Veh, vehicle diet; GHR, GH-releaser diet. aP<0.01 vs. wild-type mice treated with aCSF and vehicle diet; bP<0.05; cP<0.01 vs. klotho mutant mice treated with aCSF and vehicle diet; dP<0.05; eP<0.01 vs. klotho mutant mice treated with aCSF and GH releaser diet (three-way analysis of variance followed by multiple pair-wise comparisons with Bonferroni's correction).

• 

  Fig. 3 Effect of JB-1, an insulin-like growth factor-1 (IGF-1) receptor antagonist, on the growth hormone (GH)-releaser diet-mediated attenuation of changes in the expression of (A) phospho-Akt (p-Akt), (B) phospho-glycogen synthase kinase3β (p-GSK3β), and (C) phospho-calcium/calmodulin-dependent kinase II (p-CaMKII) in the hippocampus of klotho mutant mice. Each value is the mean±SEM of six animals. aCSF, artificial cerebrospinal fluid; Veh, vehicle diet; GHR, GH-releaser diet. aP<0.01 vs. wild-type mice treated with aCSF and vehicle diet; bP<0.01 vs. klotho mutant mice treated with aCSF and vehicle diet; cP<0.05; dP<0.01 vs. klotho mutant mice treated with aCSF and a GH-releaser diet (three-way analysis of variance followed by multiple pair-wise comparisons with Bonferroni's correction).

• 

  Fig. 4 Effect of JB-1, an insulin-like growth factor-1 (IGF-1) receptor antagonist, on the growth hormone (GH)-releaser diet-mediated attenuation of changes in the expression of (A) phospho-extracellular signal-related kinase (p-ERK), (B) phospho-c-jun N-terminal kinase (p-JNK), (C) phospho-p38 (p-p38), and (D) phospho-cAMP responsive element binding protein (p-CREB) in the hippocampus of klotho mutant mice. Each value is the mean±SEM of six animals. aCSF, artificial cerebrospinal fluid; Veh, vehicle diet; GHR, GH-releaser diet. aP<0.01 vs. wild-type mice treated with aCSF and vehicle diet; bP<0.05; cP<0.01 vs. klotho mutant mice treated with aCSF and vehicle diet; dP<0.05; eP<0.01 vs. klotho mutant mice treated with aCSF and a GH-releaser diet (three-way analysis of variance followed by multiple pair-wise comparisons with Bonferroni's correction).

• 

  Fig. 5 Effect of JB-1, an insulin-like growth factor-1 (IGF-1) receptor antagonist, on the growth hormone (GH)-releaser diet-mediated attenuation of changes in the expression of (A) Bax, (B) cleaved caspase-3, (C) Bcl-2, and (D) Bcl-xL in the hippocampus of klotho mutant mice. Each value is the mean±SEM of six animals. aCSF, artificial cerebrospinal fluid; Veh, vehicle diet; GHR, GH-releaser diet. aP<0.01 vs. wild-type mice treated with aCSF and vehicle diet; bP<0.01 vs. klotho mutant mice treated with aCSF and vehicle diet; cP<0.05; dP<0.01 vs. klotho mutant mice treated with aCSF and a GH-releaser diet (three-way analysis of variance followed by multiple pair-wise comparisons with Bonferroni's correction).

• 

  Fig. 6 Effect of JB-1, an insulin-like growth factor-1 (IGF-1) receptor antagonist, on the growth hormone (GH)-releaser diet-mediated attenuation of lipid peroxidation, as measured by (A) the malondialdehyde (MDA) level, and protein oxidation, as measured by (B) the protein carbonyl level in the hippocampus of klotho mutant mice. Each value is the mean±SEM of eight animals. DNPH, 2,4-dinitrophenylhydrazine; aCSF, artificial cerebrospinal fluid; Veh, vehicle diet; GHR, GH-releaser diet. aP<0.05; bP<0.01 vs. wild-type mice treated with aCSF and vehicle diet; cP<0.05; dP<0.01 vs. klotho mutant mice treated with aCSF and vehicle diet; eP<0.05 vs. klotho mutant mice treated with aCSF and a GH-releaser diet (three-way analysis of variance followed by multiple pair-wise comparisons with Bonferroni's correction).

• 

  Fig. 7 Effect of JB-1, an insulin-like growth factor-1 (IGF-1) receptor antagonist, on the growth hormone (GH)-releaser diet-mediated attenuation of cognitive dysfunction, as evaluated by (A) a novel object recognition test and (B) a context-dependent conditioned fear learning test in klotho mutant mice. Each value is the mean±SEM of 10 animals. aCSF, artificial cerebrospinal fluid; Veh, vehicle diet; GHR, GH-releaser diet. aP<0.01 vs. wild-type mice treated with aCSF and vehicle diet; bP<0.01 vs. klotho mutant mice treated with aCSF and vehicle diet; cP<0.05 vs. klotho mutant mice treated with aCSF and a GH-releaser diet (three-way analysis of variance followed by multiple pair-wise comparisons with Bonferroni's correction).

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