Comparative Study on Biological Activities of  Decoction According to the Preservation Periods

Seong Eun Jin; Ohn Soon Kim; Hyeun-Kyoo Shin; Soo-Jin Jeong; Jin, Seong Eun; Kim, Ohn Soon; Shin, Hyeun-Kyoo; Jeong, Soo-Jin

doi:10.13048/jkm.14031

JKM > Volume 35(3); 2014 > Article

Jin, Kim, Shin, and Jeong: Comparative Study on Biological Activities of Gwakhyangjeonggi-san Decoction According to the Preservation Periods

Original Article

Journal of Korean Medicine 2014; 35(3): 60-69.

Published online: September 30, 2014

DOI: https://doi.org/10.13048/jkm.14031

Comparative Study on Biological Activities of Gwakhyangjeonggi-san Decoction According to the Preservation Periods

Seong Eun Jin, Ohn Soon Kim, Hyeun-Kyoo Shin, Soo-Jin Jeong

Herbal Medicine Formulation Research Group, Korea Institute of Oriental Medicine

Correspondence to: 정수진 (Soo-Jin Jeong), 대전광역시 유성구 유성대로 1672 한국한의학연구원 한약연구본부 한약방제연구그룹, Tel: +82-42-868-9651, Fax: +82-42-864-2120, E-mail: sjijeong@kiom.re.kr

Received July 14, 2014 Revised September 11, 2014 Accepted September 16, 2014

Abstract

Background:

Herbal formulas are generally served as a type of decoction. However, there is no scientific evidence for determining preservation and circulation period of herbal medicine decoctions. Thus, we investigated anti-inflammatory and anti-oxidative effects of the Gwakhyangjeonggi-san decoction according to its preservation period.

Methods:

Gwakhyangjeonggi-san decoction was stored for 0, 1, 2 or 3 months at room temperature. To evaluate anti-inflammatory effects, enzyme-linked immunosorbent assays (ELISAs) for tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6), and nitric oxide (NO) assay were conducted using the culture supernatant from RAW 264.7 cells stimulated with lipopolysaccharide (LPS). The antioxidant activities were studied by measuring free radical scavenging activities on ABTS and DPPH.

Results:

Gwakhyangjeonggi-san decoction maintained the inhibitory effects on TNF-α, IL-6 and NO productions after up to 2 months of storage in LPS-treated RAW 264.7 cells. No inflammatory response was observed in 3 months of storage. In addition, the scavenging activities on ABTS and DPPH of Gwakhyangjeonggi-san decoction were reduced time-dependently and showed less than 50% inhibition after 3 months of storage.

Conclusions:

Our results suggest that preservation period of Gwakhyangjeonggi-san decoction is recommended within 2 months after storage.

Keywords: Gwakhyangjeonggi-san decoction, preservation period, anti-inflammation, anti-oxidation

Fig. 1.

Effect of Gwakhyangjeonggi-san stored with different period on LPS-induced NO production in RAW 264.7 cells. (A) Gwakhyangjeonggi-san of 0 month storage; (B) Gwakhyangjeonggi-san of 1 month storage; (C) Gwakhyangjeonggi-san of 2 month storage; (D) Gwakhyangjeonggi-san of 3 month storage. Cells were pre-treated with Gwakhyangjeonggi-san or l-NMMA (100 M) for 4 h and then stimulated with LPS (1 g/mL) for 20 h. NO production was measured in the culture medium with the Griess reaction. The data are mean values of three experiments ± SEM (n=3). ^# p<0.01 versus vehicle-treated control group; ^* p<0.05 and ^** p<0.01 versus LPS-treated cells.

Fig. 2.

Effect of Gwakhyangjeonggi-san stored with different period on LPS-induced TNF-α production in RAW 264.7 cells. (A) Gwakhyangjeonggi-san of 0 month storage; (B) Gwakhyangjeonggi-san of 1 month storage; (C) Gwakhyangjeonggi-san of 2 month storage; (D) Gwakhyangjeonggi-san of 3 month storage. Cells were pre-treated with Gwakhyangjeonggi-san for 4 h and then stimulated with LPS (1 g/mL) for 20 h. The levels of TNF-α released into the culture medium were assessed using commercially available ELISA kit. The data are mean values of three experiments ± SEM (n=3). ^# p<0.01 versus vehicle-treated control group; ^** p<0.01 versus LPS-treated cells.

Fig. 3.

Effect of Gwakhyangjeonggi-san stored with different period on LPS-induced IL-6 production in RAW 264.7 cells. (A) Gwakhyangjeonggi-san of 0 month storage; (B) Gwakhyangjeonggi-san of 1 month storage; (C) Gwakhyangjeonggi-san of 2 month storage; (D) Gwakhyangjeonggi-san of 3 month storage. Cells were pre-treated with Gwakhyangjeonggi-san for 4 h and then stimulated with LPS (1 g/mL) for 20 h. The levels of IL-6 released into the culture medium were assessed using commercially available ELISA kit. The data are mean values of three experiments ± SEM (n=3). ^# p<0.01 versus vehicle-treated control group; ^** p<0.01 versus LPS-treated cells.

Fig. 4.

Effect of Gwakhyangjeonggi-san stored with different period on ABTS radical scavenging activity. ABTS radical cation was produced by reacting 7 mM ABTS solution with 2.45 mM potassium persulfate store in the dark at room temperature for 16 h. Then, ABTS radical solution was added to a 96-well plate containing of samples or ascorbic acid (AA, 5 g/mL). After 30 min of incubation, the absorbance was measured at 743 nm using a microplate reader. The data are mean values of three experiments ± SEM (n=3). ^* p<0.1, ^** p<0.01 versus 0 month.

Fig. 5.

Effect of Gwakhyangjeonggi-san stored with different period on DPPH radical scavenging activity. DPPH radical solution (0.15 mM in ethanol) was added to a 96-well plate containing of samples or ascorbic acid (AA, 20 g/mL). After 30 min of incubation, the absorbance was measured at 517 nm using a microplate reader. The data are mean values of three experiments ± SEM (n=3). ^** p<0.01 versus 0 month.

Table 1.

Composition of Herbal Medicine in Extract of Gwakhyangjeonggi-san

Herbal medicine	Latin name	Original region	Company	Amount (g)
Agastache rugosa (Fisch. et Meyer) O. Kuntze	Agastachis Herba	Andong, Gyeongbuk, Korea	Gwangmyungdang Pharm	5.63
Perilla frutescens var. crispa (Thunb.) H. Deane	Perillae Folium	Yeongcheon, Gyeongbuk, Korea	Gwangmyungdang Pharm	3.75
Angelica dahurica Benth. et Hook. f.	Angelicae Dahuricae Radix	Uljin, Gyeongbuk, Korea	Gwangmyungdang Pharm	1.88
Areca catechu L.	Aracae Pericarpium	China	Gwangmyungdang Pharm	1.88
Poria cocos F. A. Wolf	Poria Sclerotium	Pyeongchang, Gangwon, Korea	Gwangmyungdang Pharm	1.88
Magnolia officinalis Rehd. et E. H. Wils.	Magnoliae Cortex	China	Gwangmyungdang Pharm	1.88
Atractylodes macrocephala Koidz.	Atractylodes Rhizoma Alba	China	Gwangmyungdang Pharm	1.88
Citrus reticulata Blanco	Citrus Unshius Pericarpium	Jeju, Korea	Gwangmyungdang Pharm	1.88
Pinellia ternata Breit.	Pinelliae Tuber	China	Gwangmyungdang Pharm	1.88
Platycodon grandiflorum A. DC.	Platycodonis Radix	Andong, Gyeongbuk, Korea	Gwangmyungdang Pharm	1.88
Glycyrrhiza uralensis Fisch.	Glycyrrhizae Radix et Rhizoma	China	Gwangmyungdang Pharm	1.88
Ziziphus jujuba var. inermis (Bunge) Rehder	Zizyphi Fructus	Yeongcheon, Gyeongbuk, Korea	Gwangmyungdang Pharm	3.75
Zingiber officinale Rosc.	Zingiberis Rhizoma Recens	Ulsan, Korea	Gwangmyungdang Pharm	3.75

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