대한화장품학회지 (Journal of the Society of Cosmetic Scientists of Korea)

  • 제45권1호
  • /
  • Pages.77-85
  • /
  • 2019
  • /
  • 1226-2587(pISSN)
  • /
  • 2288-9507(eISSN)
대한화장품학회 (Society of Cosmetic Scientists of Korea)
권호 표지
DOI QR코드

DOI QR Code

오미약성이론을 활용한 황련, 승마, 백복령 및 복합물의 생리활성

Physiological Activity of Coptis japonica, White Poria cocos, and Cimicifuga heracleifolia and a Mixture of Their Extracts on Skin Based on The Oriental Medicine OMiYakSung Theory

  • 유화선 ((주)코씨드바이오팜 바이오융합연구소) ;
  • 오성화 ((주)코씨드바이오팜 바이오융합연구소) ;
  • 이정노 ((주)코씨드바이오팜 바이오융합연구소) ;
  • 김희택 (세명대학교부속 제천한방병원) ;
  • 하헌용 (세명대학교부속 제천한방병원) ;
  • 김용민 (세명대학교부속 제천한방병원) ;
  • 박성민 ((주)코씨드바이오팜 바이오융합연구소)
  • Ryu, Hwa Sun (Bio Convergence R&D Center, CoSeedBioPharm Corporation) ;
  • Oh, Seong-Hwa (Bio Convergence R&D Center, CoSeedBioPharm Corporation) ;
  • Lee, Jung-No (Bio Convergence R&D Center, CoSeedBioPharm Corporation) ;
  • Kim, Hee-Taek (Department of Oriental Medical Opthalmology & Otolaryngology & Dermatology, Semyung University Oriental Medical Hospital) ;
  • Ha, Hun-Yong (Department of Oriental Medical Opthalmology & Otolaryngology & Dermatology, Semyung University Oriental Medical Hospital) ;
  • Kim, Yong-Min (Department of Oriental Medical Opthalmology & Otolaryngology & Dermatology, Semyung University Oriental Medical Hospital) ;
  • Park, Sung-Min (Bio Convergence R&D Center, CoSeedBioPharm Corporation)
  • 투고 : 2019.01.14
  • 심사 : 2019.03.20
  • 발행 : 2019.03.30
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

오미약성 이론은 다섯가지 맛 중에서 적어도 하나 이상을 맛을 포함하는 다양한 허브로 구성되어 있으며, 이러한 이론은 인간의 질병을 예방하고 면역 체계를 강화하는데 사용되어왔다. 본 연구의 목적은 오미약성 이론에 근거한 단일 추출물 및 혼합 추출물의 효능 차이와 성분 변화를 확인하는 것이다. 황련, 승마 및 백복령 세가지 약초를 선택하였고 단일 추출물 및 혼합 추출물의 약리학적 효능을 평가하였다. 결과적으로 혼합추출물은 단일추출물과 비교하여 400 ug/mL 농도에서 우수한 세포 이동 효과를 보였다. 또한 혼합추출물은 수지상세포의 활성을 증가시켜 면역을 강화시켰으며 DPPH assay 및 HPLC-ABTS assay를 통해 가장 높은 항산화 활성을 보였다. 이번 연구에서 우리는 한의학적 이론을 접목하여 화장품 및 의약품 분야에 응용 가능한 새로운 소재를 개발하였다.

OMiYakSung theory consists of various herbs that contain at least one or more of the five flavors. This theory has been used to prevent human diseases and enhance the immune system. The main objective of the present study was to investigate efficacy differences and changes in ingredients of blended and single herb extracts based on OMiYakSung theory. We selected three herbs Coptis japonica Makino, Cimicifuga heracleifolia Komarov, and white Poria cocos and assessed their physiological effect. As results, the blended extracts showed excellent cell migration effect at 400 ug/mL concentration, compared to the single extract. In addition, the blended extracts enhanced immune function by increasing the activity of dendritic cells and showed the highest antioxidant activity by DPPH assay and HPLC-ABTS assay. In this study, we developed a new materials that can be applicable to cosmetics and pharmaceuticals field by applying oriental medicine theory.

키워드

1. Introduction

Korean traditional medicine (KTM) originated thousands of years ago, and it can be traced as far back as 3000 B.C. It is used in conjunction with acupuncture, moxibustion, herbal medicine, and cupping[1]. KTM theories are primarily focused on prescriptions for immunity enhancement for human disease prevention[2]. Yin-yang and the five elements theory treats diseases and maintains health by balancing and harmony human body[3,4]. A variety of studies have been conducted on the dermatological efficacy of these blending prescriptions [5,6]. Cheongsangbangpung-tang aqueous extracts has skin regeneration, anti-wrinkle, whitening and moisturizing effects [7]. In addition, Chungsimbohyeltang has anti-tumor and the immunomodulatory effects[8]. Many cosmetic researchers have developed cosmetic brands using it’s theories. However, there is no conclusive information about these theories. OMiYakSung is a KTM theory that classifies herbal medicine into five different flavors that are sweet, salty, bitter, spicy, and sour. According to OMiYakSung theory, herbs with sweet flavor nourish the blood and purify the body while those with salty flavor activate metabolism as well as have anti-inflammatory and anti-pigmentation effects as discussed elsewhere[9,10]. Bitter flavor herbs reduce fever, reinforce the immune system, and protect the stomach[10,11]. Herbs with a spicy flavor stop diarrhea and remove waste products, and those with a sour flavor affect blood circulation, alleviate disease symptoms, and protect the liver as discussed by Kim[1]. Sweet flavor herbs contain glucose[9,12], salty flavor herbs contain inorganic compounds[9], bitter flavor herbs contain flavonoids and alkaloids[10,11], spicy flavor herbs contain lignan molecules [9,12,13], and sour flavor herbs contain organic acids[9]. The OMiYakSung theory explains that a disease may be cured through a balance of these herbs.

We studied many traditional books (e.g. Ong-Juh Gathering of Donguibogam, Bonchokangmok) on the relationship between OMiYakSung theory and skin pharmacological effects based on immunity enhancement, skin barrier, and antioxidant activity. We evaluated the cytotoxicity, anti-oxidant, and anti-inflammatory efficacy of 20 natural products selected through the traditional books. A prescription for immunity improvement and skin barrier reinforcement is comprised of three flavors, bitter, sweet, and spicy, based on the OMiYakSung theory (Ong-Juh Gathering of Donguibogam). We selected three herbs Coptis japonica (bitter flavor, CJM), white Poria cocos (sweet flavor, WPC), and Cimicifuga heracleifolia (spicy flavor, CHK), through OMiYakSung theory and efficacy. The extract of CJM, a traditional herbal medicine, has been used as an oral remedy for inflammation-related diseases[14-16], gastrointestinal cancer, gastric ulcers, and cardiovascular diseases[16]. In addition, CJM has been shown to have anti-inflammatory, anti-oxidant and skin regenerating effects on skin[17,18]. WPC has been used as adiuretic, sedative, tonic, and to treat chronic gastritis, edema, nephrosis, gastric atony, acute gastro-enteric catarrh, nauea, emesis, and dizziness[19-22]. Besides, WPC is known to have biological activities, such as anti-inflammatory and immune-modulating effects[23,24]. CHK has been used to clear heat, induce sweating to remove exopathogens and toxins, induce cardiovascular effects, and promote eruption[25-27]. Moreover, CHK has an anti-oxidant, wound healing and anti-wrinkle effect by dermatologically[28-30].

Prevention and treatment efficacy through the synergy of many active ingredients of herbs is one of the important features of KTM. It is not easy to identify scientifically where ingredients of a mixed extract act, and what kind of synergy they have. Moreover, there has been no study on the phytochemical and secondary metabolism of blended extracts. Therefore, the aim of this study was to evaluate the pharmacological activity and phytochemical difference in vitrousing biological and chemical studies. To investigate the skin pharmacological activity of the three herbs, we compared the individual herbs and a mixture of their extracts (CCW). This study assessed their antioxidants, immunity improvement, and skin regeneration properties.

2. Materials and Methods

2.1. Reagents

Three oriental herbs (Coptis japonica Maki no, Cimicifuga heracleifolia Komarov, White Poria cocos Wolf ) were purchased from Jecheon hanbangyackcho in Jecheon, Chungbuk, Korea. Dulbecco’s modified Eagle’s medium (DMEM), Ham’s Nutrient Mixture F-12 (F-12), fetal bovine serum (FBS), penicillin-streptomycin (PS), trypsin-EDTA, and phosphate-buffered saline (PBS) were purchased from Gibco (USA), ABTS, potassium persulfate for ABTS stock solution, and formic acid, trypan blue, crystal violet solution, 3-(4,5-dimethylthiazol-2–yl)- 2,5-diphenyltetrazolium bromide (MTT), DPPH, lipopolysaccharide (LPS), 2-propanol and chloroform were purchased from Sigma Chemical Co. (USA). Analytical grade acetonitrile and distilled HPLC-grade water were purchased from Fisher Scientific (USA). Dimethyl sulfoxide (DMSO), methyl alcohol (MeOH), and ethyl alcohol (EtOH) were purchased from DAEJUNG (Korea). TRIzol reagent was purchased from Invitrogen (USA). Mouse antibodies against CD11c, CD40, and CD80 were purchased from BD Biosciences (USA).

2.2. HDFn Subculture

Human dermal fibroblasts (HDFn) were purchased from ATCC (USA). They were grown in media, in which DMEM and Ham’s F-12 Nutrient Mixture were blended at a 3: 1 ratio and with 10% FBS and 1% PS solution in a humidified incubator with 5% CO2 at 37 oC. Subcultures were performed using trypsin-EDTA every 2 – 3 days.

2.3. Cell Viability

HDFn were seeded at a density of 1 × 104 cells/well in a 96 well plate. The samples were treated with a range of different herb extract concentrations and incubated for 24 h after which MTT solution was added. Formazan was dissolved by DMSO after removal of the MTT solution. The absorbance was measured at 540 nm using a microplate reader (USA).

2.4. Cell Migration

HDFn were seeded at a density of 5 × 104 cells/well in a 24 well plate. The cells were incubated for 24 h, and scratched using a 1 mm tip, and washed with PBS. The samples were diluted with a range of different herb extract concentrations. The cells were incubated for 48 h, fixed with 70% EtOH and stained with 0.2% crystal violet.

2.5. Generation of Bone-marrow (BM)-derived Dendritic Cells (DCs)

BM-derived DCs were prepared as previously described[31]. Briefly, BM cells were flushed out from femurs and tibias. After red blood cell lysis by whole BM cells (2 × 105 cells/mL) were cultured in 100-mm2 culture dishes in 10 ml/dish complete medium containing 2 ng/mL granulocyte-macrophage colony-stimulating factor (GM-CSF) (R&D Systems, USA). On day 3, another 10 mL of fresh complete medium containing 2 ng/mL GM-CSF was added, and half of the medium was changed on day 6. On day 8, non-adherent and loosely adherent DCs were harvested by vigorous pipetting and used as immature DCs. The immature DCs were generally > 85% CD11c+.

2.6. Phenotype Analysis

Cell staining was performed using a combination of fluorescein isothiocyanate (FITC)-conjugated antibodies against CD40 and CD80 with allophycocyanin (APC)-conjugated CD11c antibody. Cells were analyzed by flow cytometry (FACS Calibur, BD Biosciences, USA) and the data were analyzed using CellQuest Pro (BD Biosciences, USA). Propidium iodide (PI) was added to stain dead cells. Forward and side scatter parameters were used to gate live cells.

2.7. Online HPLC-ABTS+ Method

To test the antioxidant activity of the herbal extracts directly, the online HPLC-ABTS screening system was applied. An Agilent 1200 HPLC system (Agilent Technologies, USA), equipped with a G1312A binary pump, a G1367B auto sampler, a G1315D PDA detector, and a G1316A column oven, fitted with an additional pump to supply the ABTS radical solution was used. For the ABTS radical reagent, a 2 mM ABTS stock solution containing 3.5 mM potassium persulfate was prepared in water and diluted 8 fold in HPLC-grade water. This solution was incubated overnight in darkness at room temperature for radical stabilization. Antioxidant capacity was measured according to a previous method[32]. Acetonitrile (A) and water containing 0.1% formic acid (B) were used for the gradient solvent system with an INNO C18 column (250 × 4.6 mm I.D., 5 um particle size) to analyze the oriental herb extracts. The gradient conditions were as follows: 0 ~ 5 min, 10% A; 5 ~ 35 min, 10 ~ 100% A; 35 ~ 40 min, 100 ~ 100% A; and then return to the initial conditions. The column was kept at 25 oC during the entire sequence. The ABTS radical solution was supplied at a flow rate of 0.5 mL/min. Chromatograms were recorded at 280 nm as a positive peak, and the visible detector was set at 734 nm to measure the decrease of ABTS radicals as a negative peak. The data were analyzed by ChemStation software (Agilent Technologies, USA)[32].

2.8. Evaluation of Free Radical Scavenging Activity by The DPPH Assay

The free radical scavenging effect was assessed using the DPPH radical. A DPPH solution was prepared at 0.1 mM in MeOH. The oriental herbal extracts were mixed at various concentrations with the 0.1 mM DPPH solution and incubated without light for 10 min. Absorbance was measured using a microplate reader (MULTI SKAN, Thermo, USA) at 540 nm.

2.9. Statistical Analysis

All the data were expressed as the means ± standard deviation (SD) of at least three separate experiments performed in triplicate. The statistical analysis was performed using Microsoft Excel software (Student’s t-test, * p < 0.05).

3. Results and Discussion

3.1. Regeneration of HDFn Cells

MTT assay was performed to determine cell viability when tested against the herb extracts. CCW and WPC appeared to be nontoxic up to 250 μg/mL, whereas CHK was mildly cytotoxic at the same concentration. CJM was cytotoxic at concentrations above 125 μg/mL (Figure 1). We investigated the effect of CJM, CHK, WPC, and CCW on cell migration. The cell migration effect was dependent on the concentration of the extract, especially for CCW, which shows the most filled blank area. Therefore, CCW has excellent regeneration activity in HDFn cells (Figure 2).

HJPHBN_2019_v45n1_77_f0001.png 이미지

Figure 1. Cell viability. HDFn cells were treated with various concentrations of CJM (A), CHK (B), WPC (C) and CCW (D) for 24 h. Viability was determined in the MTT assay by measuring the absorbance at 540 nm. The results are expressed as the percentage of viability over UN ( * p < 0.05). All data are expressed as the mean ± SD of three separate experiments performed in triplicate.

HJPHBN_2019_v45n1_77_f0002.png 이미지

Figure 2. Skin regeneration effect of oriental herbs extract in HDFn cells. Cells were scratched by 1 mm tip and visualized after 0 h (A) and 48 h (B) using crystal violet staining. Cells were treated with various concentration of CJM (D, H, and L), CHK (E, I, and M), WPC (F, J, and N) and CCW (C, G, and K) for 48 h. The black line represents the scratched area. The wound width were measured using image J software (O). All data are expressed as the mean ± SD of three separate experiments performed in triplicate.

3.2. Effect of Three Herbs and CCW on Immune Improvement

DCs is a critical antigen-presenting cell that induces primary immune responses. Under normal conditions, DCs are termed immature DCs on the human body[33]. After exposure to an antigen, immature DCs mature and migrate to the secondary lymphoid organ for T cell activation[34]. Matured DCs show high expression of CD40 and CD80 and high secretion of cytokines such as IL-12[35-37]. In this study, we investigated the phenotypic maturation of DCs by determining the expression level of CD40 and CD80. DCs were treated with LPS, CCW, CJM, CHK, and WPC. CCW and CJM enhanced the expression of CD40 and CD80 (Figure 3A) in a dose-dependent manner. Although 100 μg/mL of CCW and CJM was toxic to DCs, a non-toxic dose increased the expression of surface molecules (Figure 3A, B). However, CHK and WPC did not affect the expression of cell surface molecules (Figure 3A). LPS-, CCW- and CJM-treated DCs showed a mature morphology with long dendrites, whereas CHK-, WPC-, and untreated DCs had short dendrites (Figure 3C). These results demonstrated that CCW and CJM induce phenotypic maturation of DCs, which subsequently activated the T cells.

HJPHBN_2019_v45n1_77_f0003.png 이미지

Figure 3. Phenotypic maturation of DCs by oriental herbal extract. Immature DCs were treated with LPS (1 μg/mL), CCW, CJM, CHK, and WPC. After incubation for 24 h, DCs were stained with APC-conjugated CD11c Ab plus FITC-conjugated Abs against CD40 and CD80 (A). PI was added to stain dead cells (B). Mean fluorescence intensities (MFI) of three separate experiments are shown. Significances were determined using the Student’s t-test versus chemically untreated control groups (UN) ( * p < 0.05). (C) Cell morphology change of DCs by oriental herbal extracts. All data are expressed as the mean ± SD of three separate experiments performed in triplicate.

3.3. Effect of Three Herbs and CCW on Antioxidant Activity

To investigate the antioxidant effect, we examined using the DPPH assay and Online HPLC-ABTS. Results indicated that CHK and CCW had higher antioxidant activity than the positive control, 100 mg/mL L-ascorbic acid(Figure 4). To investigate the major compounds of the oriental herbs and to measure their ABTS radical-scavenging activity, 70% EtOH reflux extracts of the herbs were subjected to the online HPLC−ABTS+ screening system. The resulting reduction signal was detected as a negative peak by an absorbance detector (at 734 nm for ABTS+ scavenging activity (Figure 5A). The online HPLC−ABTS+ assay system allowed for comparison of the increased anti-oxidant peaks that resulted from the mixing and reflux extraction of the oriental herbs, without requiring analyte isolation. The CCW chromatogram showed an increased peak at 17.05 min and 16.20 min, with anti-oxidative activity (Figure 5B, black arrow). CCW had the highest increased anti-oxidant activity compared to that of the single extracts.

HJPHBN_2019_v45n1_77_f0004.png 이미지

Figure 4. DPPH assay of oriental herb extracts. The amount of DPPH radicals was determined at various concentrations of CJM (A), CHK (B), WPC (C) and CCW (D) by a microplate reader at 540 nm. All data are expressed as the mean ± SD of three separate experiments performed in triplicate ( * p < 0.05).

 

HJPHBN_2019_v45n1_77_f0005.png 이미지

Figure 5. The total qualitative anti-oxidative activity of oriental herb extracts. (A) Anti-oxidative activities were calculated as negative peak area from the online HPLC-ABTS screening system. Anti-oxidative activity peak area; CJM (522), CHK (577), WPC (52), and CCW (822). (B) Chromatogram of the online HPLC-ABTS screening system. The 70% EtOH reflux extracts of oriental herbs, CJM, CHK, WPC, and of the mixture CCW, were analyzed with an HPLC system, and the eluates from the HPLC column were reacted with ABTS radical reagent. Analytes were detected at 280 nm as a positive peak, and their ABTS radical-scavenging activity was recorded at 734 nm as a negative peak. All data are expressed as the mean ± SD of three separate experiments performed in triplicate ( * p < 0.05).

Previous studies have isolated and identified ingredients from single herb extracts. Furthermore, there have been no reports on the synergistic effect or phytochemical validation of the blended extracts. Previous studies have only described a variety of activities and phytochemicals including secondary metabolites from single medicinal extracts. However, these studies have not compared blended and single extracts. There are many phytochemicals in medicinal herb extracts, and they have potential for organic synthesis reactions in the body because of factors such as pH, enzyme activity, and solubility. Blended extracts can synthesize new compounds through hydrolysis, substitution, addition, and elimination reaction. In this study, the blended extract method was more effective than the single extract method. The results indicate that KTM based blended extracts provide activities or new activities through changed molecular interactions. Overall, the KTM prescription for the blended extract is useful, but lacks information about its advantages and disadvantages in the field of phytochemicals and efficacy. We hope that scientific studies will be continued in the same vein as our research.

4. Conclusions

OMiYakSung is a korean traditional medicine theory that classifies herbal medicine into five different flavors that are sweet, salty, bitter, spicy, and sour. We studied many traditional books on the relationship between OMiYakSung theory and skin pharmacological effect. In this study, we found that three herbs complex extract based on the OMiYakSung theory reinforce antioxidant, immune system, and skin regeneration effect. Thus, the development of cosmetic ingredients based on the OMiYakSung theory would provide promising candidates for use in the cosmetic industry.

Acknowledgment

This research was supported by a grant of the Korea Health Technology R&D Project the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (grant number: HN13C0069).

† Hwa Sun Ryu and Seong-Hwa Oh contributed equally to this paper.

참고문헌

  1. T. Kim, Classical texts in the present tense: the looking diagnosis of a Donguibogam school in South Korea, J Altern Complement Med, 20(4), 300 (2014). https://doi.org/10.1089/acm.2012.0507
  2. J. R. Underwood, M. Chivers, T. T. Dang, and P. V. Licciardi, Stimulation of tetanus toxoid-specific immune responses by a traditional Chinese herbal medicine, Vaccine, 27(47), 6634 (2009). https://doi.org/10.1016/j.vaccine.2009.03.051
  3. H. R. Kim, The beginning and development of Wuxing theory of Huang Di Nei Jing, Eastern Classic Stid, 29, 99 (2013).
  4. G. D. Shim, General edition: reading and healing the emotions by yin-yang and the five elements theory: based on the Hwang-je-Naegyeong, Youngsan J. East Asian Cult. Stud, 16, 155 (2013). https://doi.org/10.22863/EACS.2013.16..155
  5. J. S. Lee and Y. C. Kim, The promoting effect of Angelica gigas Nakai and Glycyrrhiza uralensis Fischet oriental medicine complex extracts on hair growth, J. Cosmet. Sci., 6, 49 (2010).
  6. J. B. Lee and H. Y. Ha, Moisturizing effect of a Taklisodokeum-containing cream, J. Invest. Cosmetol, 11, 225 (2015). https://doi.org/10.15810/jic.2015.11.3.006
  7. C. Y. Woo and D. C. Kim, Skin regeneration, anti-wrinkle, whitening and moisturizing effects of Cheongsangbangpung-tang aqueous extracts with cytotoxicity, J. Oriental Gynecology, 30(2), 49 (2017).
  8. S. K. Park, B. H. Ryu, D. W. Park, and K. W. Ryu, Experimental studies on the anti-tumor and the immunomodulatory effects of Chungsimbohyeltang, J. Int. Korean Med., 19(1), 221 (1998).
  9. K. D. Matson, J. R. Millam, and K. C. Klasing, Thresholds for sweet, salt, and sour taste stimuli in cockatiels (Nymphicus hollandicus), Zoo Biol., 20(1), 1 (2001). https://doi.org/10.1002/zoo.1001
  10. H. Matsunami, J. P. Montmayeur, and L. B. Buck, A family of candidate taste receptors in human and mouse, Nature, 404(6778), 601 (2000). https://doi.org/10.1038/35007072
  11. J. P. Ley, M. Blings, S. Paetz, G. E. Krammer, and H. J. Bertram, New bitter-masking compounds: hydroxylated benzoic acid amides of aromatic amines as structural analogues of homoeriodictyol, J. Agric. Food Chem., 54(22), 8574 (2006). https://doi.org/10.1021/jf0617061
  12. J. Kalinowska-Tluscik, K. Jarzembek, J. Sliwinski, B. J. Oleksyn, V. 12Kozik, and J. Polanski, Bitter sweeteners: tetrazole derivatives of arylsulfonylalcanoids -synthesis, structure and comparative study, Acta Cryst., B64(Pt 6), 760 (2008).
  13. B. P. Bryant and I. Mezine, Alkylamides that produce tingling paresthesia activate tactile and thermal trigeminal neurons, Brain Res., 842(2), 452 (1999). https://doi.org/10.1016/S0006-8993(99)01878-8
  14. Y. Y. Choi, M. H. Kim, I. H. Cho, J. H. Kim, J. Hong, T. H. Lee, and W. M. Yang, Inhibitory effect of Coptis chinensis on inflammation in LPS-induced endotoxemia, J Ethnopharmacol, 149(2), 506 (2013). https://doi.org/10.1016/j.jep.2013.07.008
  15. M. K. Lee and H. S. Kim, Inhibitory effects of protoberberine alkaloids from the roots of Coptis japonica on catecholamine biosynthesis in PC12 cells, Planta Med., 62(1), 31 (1996). https://doi.org/10.1055/s-2006-957791
  16. H. J. Park, Y. J. Kim, K. Leem, S. J. Park, J. C. Seo, H. K. Kim, and J. H. Chung, Coptis japonica root extract induces apoptosis through caspase3 activation in SNU-668 human gastric cancer cells, Phytother Res, 19(3), 189 (2005). https://doi.org/10.1002/ptr.1539
  17. F. Liu and T.B. Ng, Antioxidative and free radical scavenging activities of selected medicinal herbs. Life Sci., 66(8), 725 (2000). https://doi.org/10.1016/S0024-3205(99)00643-8
  18. H. B. Chen, Anti-inflammatory activity of coptisine free base in mice through inhibition of NF-kappaB and MAPK signaling pathways, Eur. J. Pharmacol., 811, 222 (2017). https://doi.org/10.1016/j.ejphar.2017.06.027
  19. Y. L. Feng, P. Lei, T. Tian, L. Yin, D. Q. Chen, H. Chen, Q. Mei, Y. Y. Zhao, and R. C. Lin, Diuretic activity of some fractions of the epidermis of Poria cocos, J Ethnopharmacol, 150(3), 1114 (2013). https://doi.org/10.1016/j.jep.2013.10.043
  20. Y. T. Lu, Y. C. Kuan, H. H. Chang, and F. Sheu, Molecular cloning of a Poria cocos protein that activates Th1 immune response and allays Th2 cytokine and IgE production in a murine atopic dermatitis model, J. Agric. Food Chem., 62(13), 2861 (2014). https://doi.org/10.1021/jf405507e
  21. J. Tang, J. Nie, D. Li, W. Zhu, S. Zhang, F. Ma, Q. Sun, J. Song, Y. Zheng, and P. Chen, Characterization and antioxidant activities of degraded polysaccharides from Poria cocos sclerotium, Carbohydr Polym, 105, 121 (2014). https://doi.org/10.1016/j.carbpol.2014.01.049
  22. Y. Sun, Biological activities and potential health benefits of polysaccharides from Poria cocos and their derivatives, Int. J. Biol. Macromol., 68, 131 (2014). https://doi.org/10.1016/j.ijbiomac.2014.04.010
  23. S. J. Yu and J. Tseng, Fu-Ling, a Chinese herbal drug, modulates cytokine secretion by human peripheral blood monocytes, Int. J. Immunopharmacol., 18(1), 37 (1996). https://doi.org/10.1016/0192-0561(95)00103-4
  24. J. W. Jeong, Ethanol extract of Poria cocos reduces the production of inflammatory mediators by suppressing the NF-kappaB signaling pathway in lipopolysaccharide-stimulated RAW 264.7 macrophages, BMC Complement Altern Med, 14, 101 (2014). https://doi.org/10.1186/1472-6882-14-101
  25. Y. Da, K. Niu, K. Wang, G. Cui, W. Wang, B. Jin, Y. Sun, J. Jia, L. Qin, and W. Bai, A comparison of the effects of estrogen and Cimicifuga racemosa on the lacrimal gland and submandibular gland in ovariectomized rats, PLoS One, 10(3), e0121470 (2015). https://doi.org/10.1371/journal.pone.0121470
  26. D. Schmid, F. Woehs, M. Svoboda, T. Thalhammer, P. Chiba, and T. Moeslinger, Aqueous extracts of Cimicifuga racemosa and phenolcarboxylic constituents inhibit production of proinflammatory cytokines in LPS-stimulated human whole blood, Can. J. Physiol. Pharmacol., 87(11), 963 (2009). https://doi.org/10.1139/Y09-091
  27. E. Y. Kim, Y. J. Lee, and M. R. Rhyu, Black cohosh (Cimicifuga racemosa) relaxes the isolated rat thoracic aorta through endothelium-dependent and - independent mechanisms, J Ethnopharmacol, 138(2), 537 (2011). https://doi.org/10.1016/j.jep.2011.09.048
  28. F. Wang, S. Zhao, F. Li, B. Zhang, Y. Qu, T. Sun, T. Luo, and D. Li, Investigation of antioxidant interactions between Radix astragali and Cimicifuga foetida and identification of synergistic antioxidant compounds, PLoS One, 9(1), e87221 (2014). https://doi.org/10.1371/journal.pone.0087221
  29. A. Kusano, Y. Seyama, M. Nagai, M. Shibano, and G. Kusano, Effects of fukinolic acid and cimicifugic acids from Cimicifuga species on collagenolytic activity, Biol. Pharm. Bull., 24(10), 1198 (2001). https://doi.org/10.1248/bpb.24.1198
  30. M. K. Kim, C. Y. Bang, G. J. Yun, H. Y. Kim, Y. P. Jang, and S. Y. Choung, Anti-wrinkle effects of Seungma-Galgeun-Tang as evidenced by the inhibition of matrix metalloproteinase-I production and the promotion of type-1 procollagen synthesis. BMC Complement Altern. Med, 16, 116 (2016). https://doi.org/10.1186/s12906-016-1095-z
  31. J. Y. Kim, Y. D. Yoon, J. M. Ahn, J. S. Kang, S. K. Park, K. Lee, K. B. Song, H. M. Kim, and S. B. Han, Angelan isolated from Angelica gigas Nakai induces dendritic cell maturation through toll-like receptor 4, Int. Immunopharmacol., 7(1), 78 (2007). https://doi.org/10.1016/j.intimp.2006.08.017
  32. W. R. Kim, E. O. Kim, K. Kang, S. Oidovsambuu, S. H. Jung, B. S. Kim, C. W. Nho, and B. H. Um, Antioxidant activity of phenolics in leaves of three red pepper (Capsicum annuum) cultivars, J. Agric. Food Chem., 62(4), 850 (2014). https://doi.org/10.1021/jf403006c
  33. H. S. Kim, B. R. Shin, H. K. Lee, Y. S. Park, Q. Liu, S. Y. Kim, M. K. Lee, J. T. Hong, Y. Kim, and S. B. Han, Dendritic cell activation by polysaccharide isolated from Angelica dahurica, Food Chem. Toxicol., 55, 241 (2013). https://doi.org/10.1016/j.fct.2012.12.007
  34. B. R. Shin, H. S. Kim, M. J. Yun, H. K. Lee, Y. J. Kim, S. Y. Kim, M. K. Lee, J. T. Hong, Y. Kim, and S. B. Han, Promoting effect of polysaccharide isolated from Mori fructus on dendritic cell maturation, Food Chem. Toxicol., 51, 411 (2013). https://doi.org/10.1016/j.fct.2012.10.018
  35. E. Garzon, P. Holzmuller, R. Bras-Goncalves, P. Vincendeau, G. Cuny, J. L. Lemesre, and A. Geiger, The Trypanosoma brucei gambiense secretome impairs lipopolysaccharide-induced maturation, cytokine production, and allostimulatory capacity of dendritic cells, Infect. Immun., 81(9), 3300 (2013). https://doi.org/10.1128/IAI.00125-13
  36. J. H. Trevino-Villarreal, L. Vera-Cabrera, P. L. Valero-Guillen, and M. C. Salinas-Carmona, Nocardia brasiliensis cell wall lipids modulate macrophage and dendritic responses that favor development of experimental actinomycetoma in BALB/c mice, Infect. Immun., 80(10), 3587 (2012). https://doi.org/10.1128/IAI.00446-12
  37. H. S. Ryu, H. K. Lee, J. S. Kim, Y. G. Kim, M. Pyo, J. Yun, B. Y. Hwang, J. T. Hong, Y. Kim, and S. B. Han, Saucerneol D inhibits dendritic cell activation by inducing heme oxygenase-1, but not by directly inhibiting toll-like receptor 4 signaling, J Ethnopharmacol, 166, 92 (2015). https://doi.org/10.1016/j.jep.2015.03.020