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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
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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.

Fig. 1
Appearance of Styrax japonica
Collect fruits and leaves of Styrax japonica in july 2015 (Up). Dry Styrax japonica in shade (Down).
acup-33-2-97f1.gif
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.
acup-33-2-97f2.gif
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.
acup-33-2-97f3.gif
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.
acup-33-2-97f4.gif
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.
acup-33-2-97f5.gif
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.
acup-33-2-97f6.gif
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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