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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

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.

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.
jkm-35-3-60f1.tif
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.
jkm-35-3-60f2.tif
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.
jkm-35-3-60f3.tif
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.
jkm-35-3-60f4.tif
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.
jkm-35-3-60f5.tif
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

참고문헌

1.. Adessi C, Soto C. Converting a peptide into adrug: strategies to improve stability and bioavailability. Curr Med Chem. 2002; 9:9. 963–78.
crossref pmid

2.. 한의과대학 방제학교수 공편저. 방제학-개정증보판-. Seoul: Yeongnimsa;2003. p. 488–90.


3.. Yu C, Zhu L. Explore of immune mechanism of HUOXIANG ZHENGQI ORAL LIQUOR restrain degranulation of mast cell. Chin Traditional Patent Med. 2002; 2:120–1.


4.. Yu C, Zhu L. Experimental researches on inhibitory effect of Huoxiang Zhengqi Liquid (藿香正气水) on histamine release. Chin J Integrative Med. 2003; 9:4. 276–80.
crossref

5.. Sun JK, Kim HH, Nam CG, Koo CM. Effects of GwakHyangJungGiSan on the arterial contraction in rabbit. J Korean Oriental Intern. 2003; 24:2. 260–8.


6.. Xue X, Huang X, Gao N, Liu E, Ren L. Effects of Huoxiang Zhengqi liquid on the anti-oxidation and expression of EGFR in gastric mucosa of rats with dampness retention syndrome. Chin J Exp Traditional Med Formulae. 2012; 18:21. 230–4.


7.. Yuan Y. Study on the effect of Huoxiang Zhengqi liquid on the gastrointestinal tract in rats. J Pract Med Tech. 2007; 14:16. 2137–8.


8.. Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biol Med. 1999; 26:1231–7.
crossref pmid

9.. Moreno MI, Isla MI, Sampietro AR, Vattuone MA. Comparison of the free radical-scavenging activity of propolis from several regions of Argentina. J Ethnophamacol. 2000; 71:109–4.
crossref

10.. Ministry of Food and Drug Safety. Ministry of Food and Drug Safety Notification 2013–253. 2013.


11.. Ministry of Food and Drug Safety. Ministry of Food and Drug Safety Notification 2013–123. 2013.


12.. Ministry of Food and Drug Safety. Ministry of Food and Drug Safety Notification 2009–101. 2009.


13.. Seo CS, Kim JH, Kim SS, Lim SH, Shin HK. Evaluation of shelf-life of Bojungikgi-tang by long-term storage test. Korean J Pharmacogn. 2013; 44:2. 200–8.


14.. Seo CS, Kim JH, Lim SH, Shin HK. Establishment of shelf-Life of Ssanghwa-tang by long-term storage test. Korean J Pharmacogn. 2012; 43:3. 257–64.


15.. Seo CS, Kim JH, Lim SH, Shin HK. Estimation of shelf-life by long-term storage test of Pyungwi-san. Korean J Oriental Med Prescription. 2011; 19:1. 183–94.


16.. Son JY, Shin JW, Son CG. Stability study for herbal drug according to storage conditions and periods. J Korean Oriental Med. 2009; 30:2. 127–32.


17.. Ha H, Shin IS, Lim HS, Jeon WY, Kim JH, Seo CS, et al. Changes in anti-inflammatory effect of Pyungwi-san decoction according to the preservation temperature and period. Formular Sci. 2012; 20:2. 29–35.


18.. Kil GJ, Lim DB, Lee YJ. A study on the degraded effect of decocted Yeonkyopaedogsan over a period. Korean J Herbology. 1998; 13:1. 173–86.


19.. Kim WG, Choi YB, Lee YJ. A study on the degraded effect of decocted Injinhotang over a period. Koean J Herbology. 1998; 13:2. 14–8.


20.. Zhou X, Chan SW, Tseng HL, Deng Y, Hoi PM, Choi PS, et al. Danshensu is the major marker for the antioxidant and vasorelaxation effects of Danshen (Salvia miltiorrhiza) water-extracts produced by different heat water-extractions. Phytomed. 2012; 19:14. 1263–9.
crossref

21.. Shen B, Truong J, Helliwell R, Govindaraghavan S, Sucher NJ. An in vitro study of neuroprotective properties of traditional Chinese herbal medicines thought to promote healthy ageing and longevity. BMC Complement Altern Med. 2013; 13:373
crossref

22.. Hilmi Y, Abushama MF, Abdalgadir H, Khalid A, Khalid H. A study of antioxidant activity, enzymatic inhibition and in vitro toxicity of selected traditional Sudanese plants with anti-diabetic potential. BMC Complement Altern Med. 2014; 14:149
crossref

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