A fermented barley and soybean formula enhances skin hydration.

Original Article Journal JCBN the 1880-5086 0912-0009 Kyoto, Original 10.3164/jcbn.15-43 Society Japan ofArticle Clinical for FreeBiochemistry Radical Research and Nutrition Japan Ajcbn15-43 fermented barley and soybean formula enhances skin hydration Sein Lee,1,2,† JongEun Kim,1,2,† Sujin Suk,3,† Oh Wook Kwon,4 Gaeun Park,1,2 Taegyu Lim,1,2 Sang Gwon Seo,1,2 Jong Rhan Kim,1,2 Dae Eung Kim,5 Miyeong Lee,6 Dae Kyun Chung,6,7 Jong Eun Jeon,5 Dong Woon Cho,5 Byung Serk Hurh,5 Sun Yeou Kim8 and Ki Won Lee1,2,9,* 1

WCU Biomodulation Major and 3Interdisciplinary Program in Agricultural Biotechnology Major, Department of Agricultural Biotechnology, Advanced Institutes of Convergence Technology and 9Institute on Aging, Seoul National University, Seoul 151921, Korea 4 Graduate School of EastWest Medical Science, Kyung Hee University, Global Campus, #1732 Deogyeongdaero, Giheunggu, Yongin, Gyeonggido, 446701, Korea 5 Sempio Fermentation Research Center; #183 Osongsaengmyeong 4ro, Osongeup, Cheongwongun, Chungcheongbukdo, 363954, Korea 6 Skin Biotechnology Center, Gyeonggi Biocenter, Suwon, Korea 7 Graduate School of Biotechnology and Institute of Life Science and Resources, Kyung Hee University, Yongin 446701, Korea 8 College of Pharmacy, Gachon University, #191 Hambakmoero, Yeonsugu, Incheon 406799, Korea 2

(Received 26 March, 2015; Accepted 1 May, 2015; Published online 30 July, 2015) 9

Skin hydration is work oneJCBN of the reproduction primary beauty andof anti vided Creative stricted Copyright This 2015 the isuse, original an Commons open distribution, © 2015 access Attribution isarticle and properly distributed License, cited.aimsunder which inofany the permits medium, terms unreprothe aging treatments. Barley (Hordeum vulgare) and soybean (Glycine max) are major food crops, but can also be used as ingredients for the maintenance of skin health. We developed a natural product based skin treatment using a barley and soybean formula (BS) incorporating yeast fermentation, and evaluated its skin hydra tion effects as a dietary supplement in a clinical study. Participants ingested a placebo (n = 33) or BS (3 g/day) containing drink (n = 32) for 8 weeks. A significant increase in hydration in the BS group as compared to the placebo group was observed on the faces of subjects after 4 and 8 weeks, and on the forearm after 4 weeks. Decreases in stratum corneum (SC) thickness were also observed on the face and forearm. BS enhanced hyaluronan (HA) and skin barrier function in vitro and reduced Hyal2 expression in human dermal fibroblasts (HDF). BS also recovered ultraviolet (UV) Binduced downregulation of HA in HaCaT cells. These results suggest that BS has promising potential for development as a health functional food to enhance skin health. Key Words:

skin hydration, barley, soybean, ultraviolet B, hyaluronan

SIntroduction kin is the largest organ of the human body and the critical protective barrier between our body and the environment.(1) Skin care is considered an important priority in overall beauty care. The hydration of skin is essential for its attractive appearance, softness, complexion, protection, and helps to reinforce its barrier properties against deleterious and exogenous environmental factors.(2) Properly hydrated skin is smooth and soft, while inadequately hydrated skin appears dry and scaly, and exhibits poor barrier function.(3) Consumers are generally receptive toward functional food ingredients in skincare products.(4) Intrinsic water binding capacity and barrier function are key properties that improve with adequate skin hydration. In the extracellular matrix (ECM) of the skin, hyaluronan (HA) plays an important role in hydration due to its water-binding properties.(5) HA is a linear, high molecular-weight polysaccharide composed of a non-sulfated repeating disaccharide unit of glucuronic acid and N-acetylglucosamine.(6) HA exists ubiquitously as one of the major components of extracellular matrices of tissues, and effectively retains water, absorbing more than 1,000 times its weight. The HA content in skin accounts for approximately half the total amount of HA present in the body. HA is synthesized by hyaluronan synthase (HAS) at the inner surface of the plasma

membrane, and the newly synthesized chains are thought to be extruded into the extracellular space through pore-like structures composed of HAS and membrane phospholipids. Three different HAS genes, HAS 1–3, have been identified in mammals. However, HA is degraded by the enzyme hyaluronidase (HYAL). HYAL-1 and HYAL-2 are major components of somatic tissue and are believed to act in concert to degrade high molecularweight HA to disaccharides and tetrasaccharides.(7) Healthy skin is an effective barrier for maintaining water retention.(8) Skin barrier dysfunction can be influenced by the dysregulation of factors including aquaporin-3 (AQP3), loricrin, involucrin, and filaggrin. Profilaggrin is the precursor protein of filaggrin, and is composed of 2 to 3 repeating filaggrin monomers.(8) Profilaggrin initially decomposes to filaggrin, which is then degraded into free amino acids (FAA) which are a major component of natural moisturizing factors.(9) AQP3 represents a family of water channel proteins which are found in the basal cell layer of the epidermis, and are essential transporters of glycerol and water.(10) Ultraviolet (UV) light is one of major causes of damage to the skin barrier and induces detectable changes in the expression of these skin barrier markers. Proper ongoing maintenance of appropriate levels of epidermal hydration helps to mitigate the skin aging process. Soybeans (Glycine max) are an important plant protein source worldwide, harboring various nutrients and functional components including isoflavonoids.(11) Isoflavone in particular has been reported to exhibit bioactive properties including preventive effects toward cancers and menopausal symptoms, as well as antiinflammatory and anti-cardiovascular disease properties.(12) Similarly, barley (Hordeum vulgare) is a major food crop that is used primarily as a cereal, but also as an industrial commodity in fermented products. β-glucan, an active constituent of barley, has been reported to prevent skin problems.(13) In a previous study, we developed a fermented barley and soybean formula (BS) with enhanced isoflavone (genistein and daidzein) and β-glucan content by yeast fermentation. BS provided protection against ultraviolet (UV) B-induced photoaging effects in hairless mice.(14) In this study, we investigated the skin hydration effects of BS in a clinical trial and assessed changes in molecular markers of skin hydration in vitro. †

These authors contributed equally to this work *To whom correspondence should be addressed. Email: [emailprotected]

J. Clin. Biochem. Nutr. | September 2015 | vol. 57 | no. 2 | 156–163

Materials and Methods Chemicals and reagents. BS was prepared as described previously.(14) Primary human dermal fibroblasts (HDFs) were

isolated from the outgrowth of foreskin obtained from 7–30-yearold healthy volunteers by JH Chung’s laboratory (Seoul National University Hospital, Korea) after approval from the Institutional Review Board at Seoul National University Hospital and Seoul

Table 1. Drink preparation for clinical trial Materials

Placebo group

Experimental group

Mixing Ratio (%)

Amount per day (g)

Mixing Ratio (%)

95.24

92.24

Roasted unpolished rice extract

Fructooligosaccarides

Cereal flavor

0.3

BS Water

Lotus leaf extract

Amount per day (g)

Rice flavor

0.2

Enzyme treated stevia

0.06

Pectin

0.1

Dextrin

0.1

Sum

100

Fig. 1. Fermented barley and soybean formula (BS) enhances skin hydration in healthy subjects. (A) Clinical study design. (B) Body weight did not change significantly as a result of BS treatment. (C) and (D) BS enhances skin hydration on the face (C) and forearm (D). The clinical study was conducted as described in Materials and Methods. Skin hydration was measured using a Corneometer® CM825. Open circle = placebo group, closed circle = BS group. Data represent mean values ± SD. P values were determined with student’s t test.

S. Lee et al.

Fig. 2. BS reduces stratum corneum thickness in healthy subjects. (A) and (B) Stratum corneum images of placebo and BStreated subjects taken at the indicated time points. Quantification of stratum corneum using Dsquame® Black Tape on the face (A) and forearm (B). (C) and (D) quantification of the stratum corneum in face (C) and forearm (D) images using ImagePro® Plus software (Media Cybernetics Inc, MD). The open circle represents the placebo group and closed circle represents the BS group. Data represent mean values ± SD. P values were determined using student’s t test.

National University. HaCaT cells were purchased from CLS Cell Lines Service GmbH (Eppelheim, Germany). Dulbecco’s modified eagle medium (DMEM) was purchased from Hyclone (Long, UT). Fetal bovine serum (FBS) and β-actin antibody were bought from Sigma-Aldrich (St. Louis, MO). The AQP-3, αtubulin, Hyal2 and HAS2 antibodies was obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Antibodies against filaggrin were purchased from Millipore (Billerica, MA). 3-(4,5dimethylatiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) powder was purchased from USB Co. (Cleveland, OH). Penicillin/ streptomycin was purchased from Invitrogen (Grand Island, NY). Protein assay kits were obtained from Bio-Rad Laboratories (Hercules, CA). Clinical study with healthy volunteers. Sixty five healthy volunteers aged 25 to 60 years with dry and dark skin were recruited for the clinical study (Supplemental Table 1*). The Medical Ethics Committee of Kyung Hee University approved

the trial (KHUSBC 2014-001). The participants were randomly assigned to two groups [control (n = 33) and experimental group (n = 32)], and then randomly assigned to two subgroups within each group to examine each side of the face. Participants were given a placebo or BS-containing drink (100 ml, Table 1) for 8 weeks. Participants were banned from using certain functional cosmetics, external applicators, and dietary supplements from two weeks before the study until its conclusion. Hydration under the dead skin (10–20 μm of the stratum corneum) was examined using a Corneometer® CM825 according to the manufacturer’s instructions (Courage + Khazaka Electronic GmbH, Cologne, Germany). The amount of stratum corneum in the interior of the forearm and front cheek were examined using D-squame® Black Tape. Images of the collected dead skin samples were magnified 70X using Charm View (Moritex, Japan) and analyzed with Image-Pro® Plus software (Media Cybernetics Inc, MD).

*See online. https://www.jstage.jst.go.jp/article/jcbn/57/2/57_1543/_article/supplement 158

Fig. 3. BS enhances hyaluronan (HA) content via inhibition of HYAL2 in human dermal fibroblasts (HDF). (A) BS enhances HA content in HDFs. HDF cells were starved in serumfree media and treated with BS at the indicated concentrations for 48 h. HA concentrations were determined by ELISA as described in Materials and Methods. (B), (C), (D), (E) and (F) effects of BS on mRNA expression of an HAregulatory enzyme in HDF cells. HDF cells were starved in serumfree media and treated with BS at the indicated concentrations for 24 h. mRNA levels of HAS1 (B), HAS2 (C), HAS3, HYAL1 (E) and HYAL2 (F) were analyzed by realtime RTPCR using specific primers as described in Materials and Methods. (G) HA reduces HYAL2 protein ex pression in HDF cells. HDF cells were starved in serumfree media and treated with BS at the indicated concentrations for 48 h. HYAL2 and βactin expression were determined by Western blot analysis as described in Materials and Methods. Data represent mean values ± SD. *p

A fermented barley and soybean formula enhances skin hydration. - PDF Download Free (2024)