Analgesic Effect of  Pharmacopuncture on Formalin-Induced Pain in Rats

Mu Seob Park; Hyun Jong Lee; Yun Kyu Lee; Mu Ryeo Kim; Hae Jin Park; Jae Soo Kim

doi:10.13045/acupunct.2016020

J Acupunct Res > Volume 33(2); 2016 > Article

Park, Lee, Lee, Kim, Park, and Kim: Analgesic Effect of Styrax Japonica Pharmacopuncture on Formalin-Induced Pain in Rats

Original Article

Acupuncture 2016; 33(2): 97-108.

DOI: https://doi.org/10.13045/acupunct.2016020

Analgesic Effect of Styrax Japonica Pharmacopuncture on Formalin-Induced Pain in Rats

Mu Seob Park¹, Hyun Jong Lee¹, Yun Kyu Lee¹, Mu Ryeo Kim², Hae Jin Park², Jae Soo Kim^1,^*

¹Department of Acupuncture & Moxibustion, Meridian & Acupoint, College of Oriental Medicine, Daegu Haany University

²Department of Herbal Pharmacology, Coll. of Korean Medicine, Daegu Hanny University

^*Corresponding author: Department of Acupuncture & Moxibustion medicine, Daegu Oriental hospital of Daegu Haany University, 136, Sinchiondong-ro, Suseong-gu, Daegu, 706-828, Republic of Korea,
Tel: +82-53-770-2112, E-mail: jaice@daum.net

Received May 16, 2016 Revised May 28, 2016 Accepted June 01, 2016

See the Original "".

Abstract

Objectives:

This study was performed to investigate the analgesic effect of Styrax japonica pharmacopuncture on formalin induced pain in rats and to figure out efficient extraction method.

Methods:

The subjects were divided into 5 groups; normal group(treated with normal saline at KI03, and injected normal saline at right hindpaw after 35 minutes), control group(treated with normal saline at KI03, and injected with formalin at right hindpaw after 35 minutes), water group(treated by hot water extraction pharmacopuncture at KI03, and injected with formalin at right hindpaw after 35 minutes), ethanol group(treated with ethanol extraction pharmacopuncture at KI03, and injected with formalin at right hindpaw after 35 minutes), and ultrasound group(treated with ultrasound extraction pharmacacupuncture and injected with fromalin formalin at right hindpaw after 35 minutes). We conducted a formalin test with ultrasonic vocalization(USV), and after the test checked for substance P, Aspartate aminotransferase(AST), and Alanine aminotransferase(ALT) concentration in the blood for each of the groups.

Results:

There was a significant analgesic effect of Styrax japonica pharmacopuncture in the early phase of the formalin test, and pharmacopuncture made with an ultrasound extracting method was observed to have a better analgesic effect than other extracting methods in early phases. The substance P concentration decreased significantly in the Styrax japonica pharmacopuncture treated group and no difference was found in the AST and ALT concentration of each group.

Conclusion:

These results demonstrated that Styrax japonica pharmacopuncture had analgesic effects in noxious nociceptors stimulation. Also pharmacopuncture made with an ultrasound extracting method had a better analgesic effect than others.

Key Words: Styrax japonica; Pharmacopuncture; Analgesic; Formalin Test; Taegye(KI03).

Fig. 1

Appearance of Styrax japonica

Collect fruits and leaves of Styrax japonica in july 2015 (Up). Dry Styrax japonica in shade (Down).

Fig. 2

The flow chart of experiment

Normal: Rats group to treat by normal saline at KI03, and inject normal saline at right hindpaw after 35 minutes.

Control: Rats group to treat by normal saline at KI03, and inject 5 % formalin at right hindpaw after 35 minutes.

Water: Rats group to treat by hot water extraction pharmacopuncture at KI03, and inject 5 % formalin at right hindpaw after 35 minutes.

Ethanol: Rats group to treat by ethanol extraction pharmacopuncture at KI03, and inject 5 % formalin at right hindpaw after 35 minutes.

Ultrasound: Rats group to treat by ultrasound extraction pharmacopuncture at KI03, and inject 5 % formalin at right hindpaw after 35 minutes.

USV: Ultra sonic vocalization.

AST: Aspartate aminotransferase.

ALT: Alanine aminotransferase.

Fig. 3

Ultra sonic vocalization during the total time of formalin test

Data are presented as mean ± SE *: p<0 01 †: p<0 05 means significant difference between Normal versus Control by Student’s t-test a b c means not sharing a common letter are significantly different among the control groups by ANOVA followed by Dunnett’s test at p<0.05.

Normal: Rats group to treat by normal saline at KI03, and inject normal saline at right hindpaw after 35 minutes.

Control: Rats group to treat by normal saline at KI03, and inject 5 % formalin at right hindpaw after 35 minutes.

Water: Rats group to treat by hot water extraction pharmacopuncture at KI03, and inject 5 % formalin at right hindpaw after 35 minutes.

Ethanol: Rats group to treat by ethanol extraction pharmacopuncture at KI03, and inject 5 % formalin at right hindpaw after 35 minutes.

Ultrasound: Rats group to treat by ultrasound extraction pharmacopuncture at KI03, and inject 5 % formalin at right hindpaw after 35 minutes.

Fig. 4

Ultra sonic vocalization during two phase of formalin test

(A) Early phase (B) Late phase

Data are presented as mean ± SE *: p<0 01, means significant difference between Normal versus Control by Student’s t-test a, b, c, means not sharing a common letter are significantly different among the control groups by ANOVA followed by Dunnett’s test at p<0.05.

USV: Ultra sonic vocalization

Normal: Rats group to treat by normal saline at KI03, and inject normal saline at right hindpaw after 35 minutes.

Control: Rats group to treat by normal saline at KI03, and inject 5 % formalin at right hindpaw after 35 minutes.

Water: Rats group to treat by hot water extraction pharmacopuncture at KI03, and inject 5 % formalin at right hindpaw after 35 minutes.

Ethanol: Rats group to treat by ethanol extraction pharmacopuncture at KI03, and inject 5 % formalin at right hindpaw after 35 minutes.

Ultrasound: Rats group to treat by ultrasound extraction pharmacopuncture at KI03, and inject 5 % formalin at right hindpaw after 35 minutes.

Fig. 5

The concentrations of substance P in the blood

Data are presented as mean ± SE *: p<0 05 means significant differences between Normal versus Control by Student’s t-test a, b Means not sharing a common letter are significantly different among the control groups by ANOVA followed by Dunnett’s test at p<0.05.

Normal: Rats group to treat by normal saline at KI03, and inject normal saline at right hindpaw after 35 minutes.

Control: Rats group to treat by normal saline at KI03, and inject 5 % formalin at right hindpaw after 35 minutes.

Water: Rats group to treat by hot water extraction pharmacopuncture at KI03, and inject 5 % formalin at right hindpaw after 35 minutes.

Ethanol: Rats group to treat by ethanol extraction pharmacopuncture at KI03, and inject 5 % formalin at right hindpaw after 35 minutes.

Ultrasound: Rats group to treat by ultrasound extraction pharmacopuncture at KI03, and inject 5 % formalin at right hindpaw after 35 minutes.

Fig. 6

The expressions of aspartate aminotransferase and alanine aminotransaminase in the blood

(A) Aspartate Aminotransferase(AST), (B) Alanine Aminotransaminase(ALT).

Data are presented as mean ± SE Difference between Normal versus Control were statistically validated by Student’s t-test at p<0.05.

Difference among the control groups were statistically validated by ANOVA at p<0.05.

Normal: Rats group to treat by normal saline at KI03, and inject normal saline at right hindpaw after 35 minutes.

Control: Rats group to treat by normal saline at KI03, and inject 5 % formalin at right hindpaw after 35 minutes.

Water: Rats group to treat by hot water extraction pharmacopuncture at KI03, and inject 5 % formalin at right hindpaw after 35 minutes.

Ethanol: Rats group to treat by ethanol extraction pharmacopuncture at KI03, and inject 5 % formalin at right hindpaw after 35 minutes.

Ultrasound: Rats group to treat by ultrasound extraction pharmacopuncture at KI03, and inject 5 % formalin at right hindpaw after 35 minutes.

Table 1

The Yield of the Extracts

Extraction Methods	Meterials (g)	Extracts (g)	Yield (%)
Hot Water Extraction	400	28.5	7.1
Ethanol Extraction	200	12.4	6.2
Ultrasound Extraction	209.8	10.49	5.0

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