Limits...
The ketogenic diet suppresses the cathepsin E expression induced by kainic acid in the rat brain.

Jeong HJ, Kim H, Kim YK, Park SK, Kang DW, Yoon D - Yonsei Med. J. (2010)

Bottom Line: Immunohistochemistry with an anti-cathepsin E antibody was performed on slides of hippocampus obtained from whole brain paraffin blocks.The change in cathepsin E mRNA expression was greatest in the hippocampus.Our results showed that KA administration increased cathepsin E expression in the rat brain and its increase was suppressed by the ketogenic diet.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, College of Medicine, Kwandong University, Gangneung, Korea.

ABSTRACT

Purpose: The ketogenic diet has long been used to treat epilepsy, but its mechanism is not yet clearly understood. To explore the potential mechanism, we analyzed the changes in gene expression induced by the ketogenic diet in the rat kainic acid (KA) epilepsy model.

Materials and methods: KA-administered rats were fed the ketogenic diet or a normal diet for 4 weeks, and microarray analysis was performed with their brain tissues. The effects of the ketogenic diet on cathepsin E messenger ribonucleic acid (mRNA) expression were analyzed in KA-administered and normal saline-administered groups with semi-quantitative and real-time reverse transcription polymerase chain reaction (RT-PCR). Brain tissues were dissected into 8 regions to compare differential effects of the ketogenic diet on cathepsin E mRNA expression. Immunohistochemistry with an anti-cathepsin E antibody was performed on slides of hippocampus obtained from whole brain paraffin blocks.

Results: The microarray data and subsequent RT-PCR experiments showed that KA increased the mRNA expression of cathepsin E, known to be related to neuronal cell death, in most brain areas except the brain stem, and these increases of cathepsin E mRNA expression were suppressed by the ketogenic diet. The expression of cathepsin E mRNA in the control group, however, was not significantly affected by the ketogenic diet. The change in cathepsin E mRNA expression was greatest in the hippocampus. The protein level of cathepsin E in the hippocampus of KA-administered rat was elevated in immunohistochemistry and the ketogenic diet suppressed this increase.

Conclusion: Our results showed that KA administration increased cathepsin E expression in the rat brain and its increase was suppressed by the ketogenic diet.

Show MeSH

Related in: MedlinePlus

Semiquantitative RT-PCR analysis of cathepsin E mRNA expression in whole brain tissue. Cathepsin E expression was lower in the KA-KD group than in the KA-ND group. RT-PCR, reverse transcription polymerase chain reaction; mRNA, messenger ribonucleic acid; KA-KD, kainic acid, ketogenic diet; KA-ND, kainic acid, normal diet.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2908849&req=5

Figure 2: Semiquantitative RT-PCR analysis of cathepsin E mRNA expression in whole brain tissue. Cathepsin E expression was lower in the KA-KD group than in the KA-ND group. RT-PCR, reverse transcription polymerase chain reaction; mRNA, messenger ribonucleic acid; KA-KD, kainic acid, ketogenic diet; KA-ND, kainic acid, normal diet.

Mentions: To analyze the changes in gene expression induced by the ketogenic diet in KA-treated rats, total RNA was isolated from the whole brains of KA-ND and KA-KD rats and subjected to a microarray analysis. The mRNA expressions of kinesin family member 5B (Kif5b), cathepsin E, beta1 subunit of adenosine monophosphate (AMP)-activated protein kinase (Prkab1), apolipoprotein B editing protein (Apobec1), and general transcription factor IIa 2 (Gtf2a2) were more than 2 times lower in the ketogenic-diet group than in the normal-diet group (Table 1). In contrast, we did not observe an increase higher than 2-fold in the expression of any gene in the KA-KD group compared to the KA-ND group. Of the down-regulated genes, only cathepsin E is known to be related to epilepsy and this correlation between cathepsin E and KA prompted us to confirm the microarray data further with an RT-PCR analysis. In accordance with the results of the microarray analysis, cathepsin E mRNA expression was lower in the KA-KD rats than in the KA-ND rats in a semiquantitative RT-PCR analysis (Fig. 2).


The ketogenic diet suppresses the cathepsin E expression induced by kainic acid in the rat brain.

Jeong HJ, Kim H, Kim YK, Park SK, Kang DW, Yoon D - Yonsei Med. J. (2010)

Semiquantitative RT-PCR analysis of cathepsin E mRNA expression in whole brain tissue. Cathepsin E expression was lower in the KA-KD group than in the KA-ND group. RT-PCR, reverse transcription polymerase chain reaction; mRNA, messenger ribonucleic acid; KA-KD, kainic acid, ketogenic diet; KA-ND, kainic acid, normal diet.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2908849&req=5

Figure 2: Semiquantitative RT-PCR analysis of cathepsin E mRNA expression in whole brain tissue. Cathepsin E expression was lower in the KA-KD group than in the KA-ND group. RT-PCR, reverse transcription polymerase chain reaction; mRNA, messenger ribonucleic acid; KA-KD, kainic acid, ketogenic diet; KA-ND, kainic acid, normal diet.
Mentions: To analyze the changes in gene expression induced by the ketogenic diet in KA-treated rats, total RNA was isolated from the whole brains of KA-ND and KA-KD rats and subjected to a microarray analysis. The mRNA expressions of kinesin family member 5B (Kif5b), cathepsin E, beta1 subunit of adenosine monophosphate (AMP)-activated protein kinase (Prkab1), apolipoprotein B editing protein (Apobec1), and general transcription factor IIa 2 (Gtf2a2) were more than 2 times lower in the ketogenic-diet group than in the normal-diet group (Table 1). In contrast, we did not observe an increase higher than 2-fold in the expression of any gene in the KA-KD group compared to the KA-ND group. Of the down-regulated genes, only cathepsin E is known to be related to epilepsy and this correlation between cathepsin E and KA prompted us to confirm the microarray data further with an RT-PCR analysis. In accordance with the results of the microarray analysis, cathepsin E mRNA expression was lower in the KA-KD rats than in the KA-ND rats in a semiquantitative RT-PCR analysis (Fig. 2).

Bottom Line: Immunohistochemistry with an anti-cathepsin E antibody was performed on slides of hippocampus obtained from whole brain paraffin blocks.The change in cathepsin E mRNA expression was greatest in the hippocampus.Our results showed that KA administration increased cathepsin E expression in the rat brain and its increase was suppressed by the ketogenic diet.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, College of Medicine, Kwandong University, Gangneung, Korea.

ABSTRACT

Purpose: The ketogenic diet has long been used to treat epilepsy, but its mechanism is not yet clearly understood. To explore the potential mechanism, we analyzed the changes in gene expression induced by the ketogenic diet in the rat kainic acid (KA) epilepsy model.

Materials and methods: KA-administered rats were fed the ketogenic diet or a normal diet for 4 weeks, and microarray analysis was performed with their brain tissues. The effects of the ketogenic diet on cathepsin E messenger ribonucleic acid (mRNA) expression were analyzed in KA-administered and normal saline-administered groups with semi-quantitative and real-time reverse transcription polymerase chain reaction (RT-PCR). Brain tissues were dissected into 8 regions to compare differential effects of the ketogenic diet on cathepsin E mRNA expression. Immunohistochemistry with an anti-cathepsin E antibody was performed on slides of hippocampus obtained from whole brain paraffin blocks.

Results: The microarray data and subsequent RT-PCR experiments showed that KA increased the mRNA expression of cathepsin E, known to be related to neuronal cell death, in most brain areas except the brain stem, and these increases of cathepsin E mRNA expression were suppressed by the ketogenic diet. The expression of cathepsin E mRNA in the control group, however, was not significantly affected by the ketogenic diet. The change in cathepsin E mRNA expression was greatest in the hippocampus. The protein level of cathepsin E in the hippocampus of KA-administered rat was elevated in immunohistochemistry and the ketogenic diet suppressed this increase.

Conclusion: Our results showed that KA administration increased cathepsin E expression in the rat brain and its increase was suppressed by the ketogenic diet.

Show MeSH
Related in: MedlinePlus