Korean J Physiol Pharmacol.  2013 Dec;17(6):505-510. 10.4196/kjpp.2013.17.6.505.

Electroacupuncture Analgesia Is Improved by Adenoviral Gene Transfer of Dopamine Beta-hydroxylase into the Hypothalamus of Rats

• Kim SJ
• Chung ES
• Lee JH
• Lee CH
• Kim SK
• Lee HJ
• Bae H

• KMID: 2285477
• DOI: http://doi.org/10.4196/kjpp.2013.17.6.505

Abstract

Electroacupuncture (EA) is a modified form of acupuncture that utilizes electrical stimulation. We previously showed that EA stimulated rats were divided into responders that were sensitive to EA and non-responders that were insensitive to EA based on the tail flick latency (TFL) test. The dopamine beta-hydroxylase (DBH) gene was more abundantly expressed in the hypothalamus of responder rats than non-responder rats. To determine whether overexpression of DBH gene expression in the hypothalamus modulate EA analgesia, we constructed a DBH encoding adenovirus and which was then injected into the hypothalamus of SD rats. Microinjection of DBH or control GFP virus into the hypothalamus had no changes on the basal pain threshold measured by a TFL test without EA treatment. However, the analgesic effect of EA was significantly enhanced from seven days after microinjection of the DBH virus, but not after injection of the control GFP virus. DBH expression was significantly higher in the hypothalamus of DBH virus injected rat than control GFP virus or PBS injected rats. Moreover, expression of the DBH gene did not affect the body core temperature, body weight, motor function or learning and memory ability. Although the functional role of DBH in the hypothalamus in the analgesic effect of EA remains unclear, our findings suggest that expression of the DBH gene in the hypothalamus promotes EA analgesia without obvious side-effects.

Keyword

Adenovirus; Analgesia; Dopamine-beta-hydroxylase; Electroacupuncture; Hypothalamus

MeSH Terms

Acupuncture
Adenoviridae
Analgesia*
Animals
Body Temperature
Dopamine beta-Hydroxylase*
Dopamine*
Electric Stimulation
Electroacupuncture*
Gene Expression
Hypothalamus*
Learning
Memory
Microinjections
Pain Threshold
Rats*
Viruses
Dopamine
Dopamine beta-Hydroxylase

Figure

• 

  Fig. 1 Effects of adenoviral gene transfer of DBH on the basal pain threshold and EA analgesia. (A) Basal TFL values in the DBH virus-injected and GFP control virus-injected rats at -1, 3, 7 and 14 days after viral injection. Data are presented as the mean±SEM. p>0.05, DBH versus GFP group at each day as determined by the unpaired t-test; p>0.05, -1 day TFL versus 3, 7 and 14 days TFL in either group as determined by one-way ANOVA. (B) The increase ratio of EA-induced analgesic effect at 3, 7 and 14 days following viral injection, compared to the EA effect prior to viral injection. *p<0.05 and ***p<0.001, between the two groups as determined by the unpaired t-test. The data are the means±S.E.M (n=16/group).

• 

  Fig. 2 Verification of the correct injection and infection of the adenovirus into the hypothalamus. Tissues were prepared 15 days after PBS or virus injection. (A) Representative confocal microphotographs of GFP fluorescence in the hypothalamus from DBH virus injected rats. 3v, 3rd ventricle. Scale bar, 200 µm. (B) Representative photographs of the Nissle staining following microinjection of the viral suspension. Arrow indicates needle track. (C) Western blot analysis showing significantly increase of DBH in the DBH virus-injected rat compared to PBS or GFP control virus injected rats at 7 days later virus injection. (D) High magnification of B.

• 

  Fig. 3 Effects of adenoviral gene transfer of DBH on body temperature, body weight, and learning and memory ability. The body temperature (A) and body weight (B) of the DBH virus-injected and the GFP control virus-injected rats at -1, 3 and 7 days following viral injection (n=7~8/group). (C) The learning and memory ability in the passive avoidance test at 7 days after viral injection. The data shown are the means±S.E.M (n=7~8/group).

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