Yonsei Med J.  2017 Jan;58(1):131-138. 10.3349/ymj.2017.58.1.131.

Hyperbaric Oxygen Pretreatment Improves Cognition and Reduces Hippocampal Damage Via p38 Mitogen-Activated Protein Kinase in a Rat Model

• Zhao B
• Pan Y
• Wang Z
• Xu H
• Song X

• KMID: 2374198
• DOI: http://doi.org/10.3349/ymj.2017.58.1.131

Abstract

PURPOSE
To investigate the effects of hyperbaric oxygen (HBO) pretreatment on cognitive decline and neuronal damage in an Alzheimer's disease (AD) rat model.
MATERIALS AND METHODS
Rats were divided into three groups: normal saline (NS), AD, and HBO+AD. In the AD group, amyloid β peptide (Aβ)₁₋₄₀ was injected into the hippocampal CA1 region of the brain. NS rats received NS injection. In the HBO+AD group, rats received 5 days of daily HBO therapy following Aβ₁₋₄₀ injection. Learning and memory capabilities were examined using the Morris water maze task. Neuronal damage and astrocyte activation were evaluated by hematoxylin-eosin staining and immunohistochemistry, respectively. Dendritic spine density was determined by Golgi-Cox staining. Tumor necrosis factor-α, interleukin-1β, and interleukin-10 production was assessed by enzyme-linked immunosorbent assay. Neuron apoptosis was evaluated by terminal deoxynucleotidyl transferase dUTP nick end labeling. Protein expression was examined by western blotting.
RESULTS
Learning and memory dysfunction was ameliorated in the HBO+AD group, as shown by significantly lower swimming distances and escape latency, compared to the AD group. Lower rates of neuronal damage, astrocyte activation, dendritic spine loss, and hippocampal neuron apoptosis were seen in the HBO+AD than in the AD group. A lower rate of hippocampal p38 mitogen-activated protein kinase (MAPK) phosphorylation was observed in the HBO+AD than in the AD group.
CONCLUSION
HBO pretreatment improves cognition and reduces hippocampal damage via p38 MAPK in AD rats.

Keyword

Hyperbaric oxygen; astrocytes; TNF-α; cognitive dysfunction; p38 MAPK

MeSH Terms

Alzheimer Disease/*therapy
Amyloid beta-Peptides/*administration & dosage
Animals
Apoptosis
*Cognition/drug effects
Disease Models, Animal
Enzyme-Linked Immunosorbent Assay
Hippocampus/*enzymology
*Hyperbaric Oxygenation
In Situ Nick-End Labeling
Interleukin-10/biosynthesis
Interleukin-1beta/biosynthesis
Learning/drug effects
Male
Memory/drug effects
Neurons
Peptide Fragments/*administration & dosage
Rats
Rats, Sprague-Dawley
Sodium Chloride/administration & dosage
Tumor Necrosis Factor-alpha/biosynthesis
p38 Mitogen-Activated Protein Kinases/*metabolism
Amyloid beta-Peptides
Interleukin-10
Interleukin-1beta
Peptide Fragments
Sodium Chloride
Tumor Necrosis Factor-alpha
p38 Mitogen-Activated Protein Kinases

Figure

• 

  Fig. 1 Learning and memory ability in rats of the three experimental groups. The Morris water maze test was conducted before or on postoperative days 7, 14, and 21. The swimming distance (A), escape latency (B), and average velocity (C) were recorded in the NS, AD, and AD+HBO groups (D). The track path was recorded on postoperative day 21. n=8. *p<0.05 compared with NS, †p<0.05 compared with AD. NS, normal saline; AD, Alzheimer’s disease; HBO, hyperbaric oxygen.

• 

  Fig. 2 Neuronal damage and astrocyte activation in the hippocampal CA1 region. (A) Sections derived from the NS, AD, or AD+HBO group on postoperative day 21 were stained with hematoxylin and eosin for histological examination (upper panels). Some of the samples were stained with anti-GFAP antibody. The immunopositive cells are indicated by arrows. Magnification, 400×. (B) The percentage of activated astrocytes was calculated. (C) The TNF-α and IL-1β levels in the hippocampus were examined by ELISA. n=4. *p<0.05 compared with NS, †p<0.05 compared with AD. NS, normal saline; AD, Alzheimer’s disease; HBO, hyperbaric oxygen; TNF-α, tumor necrosis factor-α; IL, interleukin; ELISA, enzyme-linked immunosorbent assay.

• 

  Fig. 3 Dendritic spine density in the hippocampal CA1 region. (A) Sections derived from the NS, AD, or AD+HBO group on postoperative day 21 were stained with Golgi-Cox. Magnification, 1000×. (B) The number of dendritic spines per 10 µm was calculated. Ten neurons from each animal were included in the calculation, and two animals from each group were used for analysis. *p<0.05 compared with NS, †p<0.05 compared with AD. NS, normal saline; AD, Alzheimer’s disease; HBO, hyperbaric oxygen.

• 

  Fig. 4 Cell apoptosis in the hippocampal CA1 region. (A) Sections derived from the NS, AD, or AD+HBO group on postoperative day 21 were analyzed by TUNEL assay. Representative images are presented. Magnification, 400×. (B) The percentage of apoptotic cells was calculated. (C) Protein expression of cleaved caspase 3. β-actin was used as a loading control. (D) The relative expression of cleaved caspase 3 was calculated. n=4. *p<0.05 compared with NS, †p<0.05 compared with AD. NS, normal saline; AD, Alzheimer’s disease; HBO, hyperbaric oxygen; TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling.

• 

  Fig. 5 p38 MAPK phosphorylation in the hippocampus. (A) Proteins were extracted from the NS, AD, or AD+HBO group on postoperative day 21. Western blotting analysis was conducted using anti-phosphorylated p38 MAPK antibody. Representative results are presented. (B) The relative expression of phosphorylated p38 MAPK was calculated. n=4. *p<0.05 compared with NS, †p<0.05 compared with AD. NS, normal saline; AD, Alzheimer’s disease; HBO, hyperbaric oxygen; MAPK, mitogen-activated protein kinase.

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