• Title/Summary/Keyword: Mouse skin

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Development and Characterization of an Anti-Acne Gel Containing Siamese Crocodile (Crocodylus siamensis) Leukocyte Extract

  • Phupiewkham, Weeraya;Lu, Qiumin;Payoungkiattikun, Wisarut;Temsiripong, Threeranan;Jangpromma, Nisachon;Lai, Ren;Klaynongsruang, Sompong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.707-717
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    • 2018
  • Leukocytes are reportedly the first line of the innate immune defense and essential for the control of common bacterial infections. Therefore, in this work, the antibacterial activity of crocodile leukocyte extract against Propionibacterium acnes was evaluated, and we also characterized the related activity of skin infection. The leukocyte extract showed the minimum inhibitory concentration to be $100{\mu}g/ml$ to P. acnes. SEM imaging demonstrated that the leukocyte extract adversely affected P. acnes cell permeability in a concentration-dependent manner. Furthermore, the crocodile leukocyte extract could significantly reduce proinflammatory markers and decrease inflammatory signs in infected mouse ears. The crude leukocyte extract was further purified using FPLC and RP-HPLC. The resulting fraction F5 was indicated as the anti-acne peptide-containing fraction. The molecular mass of the peptide contained in F5 was calculated to be 4,790.5 Da. N-Terminal sequencing revealed the amino acid sequence as GPEPVPAIYQ, which displays similarities to immunoglobulin A and leucine-rich repeat neuronal protein. This is the first reported amino acid sequence of a crocodile leukocyte extract that possesses anti-acne activity. To attempt to use it in a prototype cosmetic, an anti-acne gel containing crude crocodile leukocyte extract was formulated, resulting in seven gel formulations (G1, G2, G3, G4, G5, G6, and G7). The formulations G5, G6, and G7 exhibited 2-fold higher anti-acne activity than G1-G4. Investigation of accelerating stability studies of anti-acne gel formulations G5, G6, and G7 demonstrated that a low storage temperature ($4^{\circ}C$) is suitable for maintaining the physical properties and biological activity of the anti-acne gel products.

The Inhibitory Effect of Chlorophyllin is Influenced by Different Promotion Stages in DMBA-TPA-induced Mouse Skin Carcinogenesis

  • Kim, Jin;Yook, Jong-In;Park, Kwang-Kyun;Lee, Eun-Ha;Jung, So-Young;Joon, Yin-Liu;Kyung, Chul-Hong;Kim, Ju;Chung, Won-Yoon
    • Environmental Mutagens and Carcinogens
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    • v.19 no.1
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    • pp.46-55
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    • 1999
  • To develop a chemopreventive strategy based on the different stages of premalignant lesions, we hypothesized that the inhibitory effect of chemopreventive agents is influenced by different promotion stages during carcinogenesis. DMBA-TPA-induced skin carcinogenesis was used with ICR mice and chlorophyllin (CHL) was applied as a chemopreventive agent. In vitro assay was performed with Salmonella typhi. TA100 to observe any anti-mutagenic activity of CHL against DMBA. Pre-initiation and pre-promotion effects of CHL were observed by CHL treatment before initiation and before promotion. To evaluate the inhibitory effect at different promotion stages, each group was divided into three subgroups after TPA promotion for 6, 18 and 24 weeks, respectively ; the first subgroup was immediately sacrificed after termination of TPA, the second subgroup was treated with CHL, and the third subgroup was sacrificed 8 weeks after termination of TPA without CHL treatment. The degrees of epithelial dysplasia, papilloma formation, and invasive carcinoma were observed histologically, and GST-Pi expression was observed immunohistochemically. ODC mRNA level was analyzed by reverse transcriptase-polymerase chain reaction. Results showed : CHL dose-dependently inhibited the mutation of Salmonella typhi. TA100; the incidence of epithelial dysplasia and papilloma formation was lower in pre-initiation and pre-promotion CHL-treated mice than DMBA-TPA-treated mice; no invasive carcinoma developed in pre-initiation CHL-treated groups, while 67% of DMBA-TPA treated mice had carcinomas; GST-Pi expression decreased when CHL was treated before initiation and before promotion; and when CHL was treated after termination of TPA application at 18 and 24- week-TPA promotion stages, respectively, the incidence of epithelial dysplasia and papilloma was markedly reduced compared to non-treated groups. When comparing the incidence of epithelial dysplasia and papilloma between the immediately-sacrificed subgroup and the non-treated group with a waiting period, we speculated that the 18-week-TPA promotion stage might belong to the promoter-independent progression stage. At the 18-week-TPA promotion stage, the level of ODC mRNA in the CHL-treated group was clearly reduced to the level of normal tissue. Taking these results together, CHL showed both anti-initiation and anti-promotion effects, while the inhibitory effect of CHL was prominent in the 18-week-TPA promotion stage. However, CHL seems to be incapable of completely blocking the progression in the 24-week-TPA promotion stage.

The Effects of Scutellaria baicalensis GEORGI on Allergic contact dermatitis induced by DNCB in Mice (황금(黃芩) 추출물(抽出物)이 DNCB로 유발(誘發)된 생쥐의 Allergy성(性) 접촉피부염(接觸皮膚炎)에 미치는 영향(影響))

  • Jun, Jae-Hong;Kang, Yoon-Ho
    • The Journal of Dong Guk Oriental Medicine
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    • v.7 no.1
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    • pp.119-133
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    • 1998
  • This study was performed to investigated the effects of an immunosuppression and a mitigation of inflammation of Scutellaria baicalensis GEORGI on Allergic contact dermatitis. In order to study, after oral administration of Scutellaria baicalensis GEORGI extract, contact hypersensitivity assay, general morphologic change of skin, sulfated acid mucosubstance, mast cell, IL-2R, ICAM-1 and CD11b were observed in BALB/C mouse induced allergic contact dermatitis by the contact sensitizing DNCB. Ear swelling in contact hypersensitivity assay was decreased in the Scutellaria baicalensis GEORGI extracts administered(HEGT) group, compared with DNCB painted group. In the general morphologic change of skin, hyperplasia of keratinocytes, increase of lymphocytes and inflammatory cell, increase of vasculogenesis and epidermal lymphocytes infiltration, increase of cell damage in epidermal basal layer and prickle layer were decreased in the HEGT group, compared with DNCB painted group. Increase of sulfated acid mucosubstance, mast cell, IL-2R positive cell, ICAM-1 positive cell, CD11b positive cell were decreased in the HEGT group, compared with DNCB painted group.

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A Potent Tissue Destructive Activity of Secreted Proteins of Aeromonas hydrophila (조직 괴사 활성을 지닌 Aeromonas hydrophila 의 분비 단백질에 관한 연구)

  • Kim, Kyu Lee;Choe, Yunjeong;Kang, Ho Young
    • Journal of Life Science
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    • v.25 no.2
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    • pp.214-222
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    • 2015
  • Aeromonas hydrophila is the most common water fish pathogen and cause diseases such as hemorrhagic septicemia, dropsy, ulceration and asymptomatic septicemia. A. hydrophila secretes many extracellular products (ECPs) which contribute to effective infection, wide distribution and great adaptability to environmental changes. Crude ECPs of A. hydrophila CK257, a strain used in this study, exhibits a toxic activity to the animals including mouse, rabbit and fish. Toxic symptoms were indicated by tissue damage and skin injuries in animal. When ECPs were subcutaneously injected to animals, skin damages were observed, appearing like necrosis. Preliminary research demonstrated that the active factors are protein component. The crude ECPs were collected after ammonium sulfate precipitation of cell-free culture supernatant. ECPs were fractionated with the use gel filtration chromatography. Five ECP fractions were obtained, of which one fraction was found to be toxic to goldfish. MALDI-TOF analyses provided two interesting proteases called M35 and M28. Both M35 and M28 are known as metalloprotease. Accordingly, proteins in an active fraction exhibited caseinolytic activity. These proteins were difference of caseinolytic activity under different metallic ions. Also active fraction has elastolytic activity. These results suggested that peptidase M28 and M35 may be a candidate factor for tissue necrosis activity about infection with A. hydrophila.

Iontophoretic Delivery of Levodopa: Permeation Enhancement by Oleic Acid Microemulsion and Ethanol (Levodopa의 이온토포레시스 경피전달: 올레인산 아이크로에멀젼 및 에탄올의 투과증진)

  • Jung, Shin-Ae;Gwak, Hye-Sun;Chun, In-Koo;Oh, Seaung-Youl
    • Journal of Pharmaceutical Investigation
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    • v.38 no.6
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    • pp.373-380
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    • 2008
  • In order to develop optimal formulation and iontophoresis condition for the transdermal delivery of levodopa, we have evaluated the effect of two permeation enhancers, ethanol and oleic acid in microemulsion, on transdermal delivery of levodopa. In vitro flux studies were performed at $33^{\circ}C$, using side-by-side diffusion cell and full thickness hairless mouse skin. Current density applied was $0.4\;mA/cm^2$ and current was off after 6 hours application. Levodopa was analysed by HPLC at 280 nm. The o/w microemulsions of oleic acid in buffer solution (pH 2.5 & 4.5) were prepared using oleic acid, Tween 80 and ethanol. The existence of microemulsion regions were investigated in pseudo-ternary phase diagrams. Contrary to our expectation, cumulative amount of levodopa transported from microemulsion (pH 2.5) for 10 hours was similar to that from aqueous solution in all delivery methods (passive, anodal and cathodal). When pH of the micro-emulsion was pH 4.5, cumulative amount of levodopa transported for 10 hours increased about 40% (anodal) to 50% (cathodal), when compared to that from aqueous solution. Flux from pH 4.5 microemulsion showed higher value than that from pH 2.5 in all delivery methods. These results seem to indicate that electroosmosis plays more dominant role than electrorepulsion in the flux of levodopa at pH 2.5. The effect of ethanol on iontophoretic flux was studied using pH 2.5 phosphate buffer solution containing 3% or 5% (v/v) ethanol. Flux enhancement was observed in passive and anodal delivery as the concentration of the ethanol increased. Without ethanol, cathodal delivery showed higher flux than anodal delivery. Anodal delivery increased the cumulative amount of levodopa transported 1.6 fold by 5% ethanol after 10 hours. However, in cathodal delivery, no flux enhancement of levodopa was observed during current application and only marginal increase in cumulative amount transported after 10 hours was observed by 5% ethanol. These results seem to be related to the decrease in dielectric constant of the medium and the lipid extraction of the ethanol, which decrease the electroosmotic flow, and thus decrease the flux. Overall, the results provide important insights into the role of electroosmosis and electrorepulsion in the transport of levodopa through skin, and provide some useful informations for optimal formulation for levodopa.

Cnestis palala (Lour.) Merr. extract suppresses Propionibacterium acnes-induced inflammation (Propionibacterium acnes에 의해 유도되는 염증반응에서 Cnestis palala (Lour.) Merr. 추출물의 억제효과)

  • Shin, Jin Hak;Lee, Eun Hye;Kim, Seon Sook;Sydara, Kongmany;Seo, Su Ryeon
    • Korean Journal of Microbiology
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    • v.54 no.1
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    • pp.38-45
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    • 2018
  • Acne is an inflammatory skin disease that occurs in puberty and young people. Propionibacterium acnes (P. acnes) is known to be a major cause of inflammation in acne. P. acnes proliferates within hair follicles blocked by overproduced sebum in the skin, and thereby activates monocytic cells to promote the secretion of pro-inflammatory cytokines. In this study, we investigated the possibility of Cnestis palala (Lour.) Merr. extract to diminish P. acnes-mediated inflammatory responses. We found that C. palala extract significantly attenuated P. acnes-induced pro-inflammatory cytokine expressions, such as $IL-1{\beta}$, IL-6, $TNF-{\alpha}$, iNOS, and COX-2 in mouse macrophage RAW264.7 cells. Moreover, we observed that C. palala extract inhibited $NF-{\kappa}B$ transcriptional activation, which is the major transcription factor of inflammatory cytokine expression. Therefore, it is expected that C. palala extract has a potential as a therapeutic agent or supplement for the treatment P. acnes-induced inflammatory responses.

Glycosyltransformation of ginsenoside Rh2 into two novel ginsenosides using recombinant glycosyltransferase from Lactobacillus rhamnosus and its in vitro applications

  • Wang, Dan-Dan;Kim, Yeon-Ju;Baek, Nam In;Mathiyalagan, Ramya;Wang, Chao;Jin, Yan;Xu, Xing Yue;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.45 no.1
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    • pp.48-57
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    • 2021
  • Background: Ginsenoside Rh2 is well known for many pharmacological activities, such as anticancer, antidiabetes, antiinflammatory, and antiobesity properties. Glycosyltransferases (GTs) are ubiquitous enzymes present in nature and are widely used for the synthesis of oligosaccharides, polysaccharides, glycoconjugates, and novel derivatives. We aimed to synthesize new ginsenosides from Rh2 using the recombinant GT enzyme and investigate its cytotoxicity with diverse cell lines. Methods: We have used a GT gene with 1,224-bp gene sequence cloned from Lactobacillus rhamnosus (LRGT) and then expressed in Escherichia coli BL21 (DE3). The recombinant GT protein was purified and demonstrated to transform Rh2 into two novel ginsenosides, and they were characterized by nuclear magnetic resonance (NMR) techniques and evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide assay. Results: Two novel ginsenosides with an additional glucopyranosyl (6→1) and two additional glucopyranosyl (6→1) linked with the C-3 position of the substrate Rh2 were synthesized, respectively. Cell viability assay in the lung cancer (A549) cell line showed that glucosyl ginsenoside Rh2 inhibited cell viability more potently than ginsenoside Rg3 and Rh2 at a concentration of 10 μM. Furthermore, glucosyl ginsenoside Rh2 did not exhibit any cytotoxic effect in murine macrophage cells (RAW264.7), mouse embryo fibroblasts cells (3T3-L1), and skin cells (B16BL6) at a concentration of 10 μM compared with ginsenoside Rh2 and Rg3. Conclusion: This is the first report on the synthesis of two novel ginsenosides, namely, glucosyl ginsenoside Rh2 and diglucosyl ginsenoside Rh2 from Rh2 by using recombinant GT isolated from L. rhamnosus. Moreover, diglucosyl ginsenoside Rh2 might be a new candidate for treatment of inflammation, obesity, and skin whiting, and especially for anticancer.

Therapeutic Effects of LED Fusion of Two Wavelength Bands on Atopic Dermatitis of NC/Nga Mice (융합 LED 광선치료가 아토피 피부염에 미치는 영향)

  • Lee, Sangmin;Choi, Ji-Hye;Koo, Bon-Jun;Kwon, Jungkee
    • Journal of the Korean Applied Science and Technology
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    • v.39 no.4
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    • pp.552-559
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    • 2022
  • Atopic dermatitis (AD) is a chronic inflammatory skin disease accompanied by severe itching, mainly before five. The aim of this study is to investigate the effects of 405 nm+850 nm LED light therapy on AD-like symptoms in NC/Nga mice. The mice were randomly placed in the normal (Vehicle), atopic dermatitis-induced (CON), and 405 nm + 850 nm LED light therapy (LED) groups. The LED experimental group conducted 405 nm+850 nm wavelength LED ray therapy for 10 minutes a day for seven days. LED light therapy research confirmed the improvement and improvement of Dermatics score and observed the reduction of epidermal tissue thickness caused by dermatitis. Based on the significant decrease of serum IL-1𝛽 and transdermal moisture loss and serum IgE concentration due to LED light therapy, LED light therapy can help restore normal skin conditions in mice that cause atopic dermatitis. This study showed the anti-atopic effect of infrared light and blue light. Light in mice with atopic dermatitis led to the simultaneous use of circular LEDs with two wavelengths.

Inhibitory Effects of Myagropsis myagroides Ethanol Extract on 2,4-Dinitrochlorobenzene-Induced Atopic Dermatitis-Like Skin Lesions in Mice (2,4-Dinitrochlorobenzene 유도 아토피 피부염 모델 마우스에 대한 외톨개 모자반(Myagropsis myagroides) 에탄올 추출물의 억제 효과)

  • Kim, Koth-Bong-Woo-Ri;Kang, Bo-Kyeong;Ahn, Na-Kyung;Choi, Yeon-Uk;Bae, Nan-Young;Park, Ji-Hye;Park, Sun-Hee;Kim, Min-Ji;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.8
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    • pp.1121-1127
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    • 2015
  • This study investigated the effects of Myagropsis myagroides ethanol extract (MMEE) on 2,4-dinitrochlorobenzene (DNCB)-induced atopic dermatitis (AD)-like skin lesions in BALB/c mice. The effects of MMEE on DNCB-induced BALB/c mice were evaluated by examining skin symptom severity, levels of total immunoglobulin E (IgE), tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), interleukin (IL)-4, and IL-10 in serum, and levels of IL-4, IL-5, IL-13, and $interferon-{\gamma}$ ($IFN-{\gamma}$) in splenocytes. MMEE significantly reduced the total clinical severity score, total IgE levels, as well as $TNF-{\alpha}$ and IL-4 production in an AD mouse model but increased IL-10 production. Production of IL-4, IL-5, and IL-13 in splenocytes was reduced by MMEE, whereas $IFN-{\gamma}$ production increased. These results suggest that MMEE can inhibit the development of AD-like skin lesions in BALB/c mice by modulating the immune response and may be an effective potential therapeutic agent for AD.

Melanogenesis regulatory constituents from Premna serratifolia wood collected in Myanmar

  • WOO, SO-YEUN
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.21-22
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    • 2019
  • Melanin is a mixture of pigmented biopolymers synthesized by epidermal melanocytes that determine the skin, eye, and hair colors. Melanocytes produce two different kinds of melanin, eumelanin (dark brown/black insoluble pigments found in dark skin and dark hair and pheomelanin (lighter red/yellow). The biological role of melanin is to prevent skin damage by ultraviolet (UV) radiation. However, the overproduction or deficiency of melanin synthesis could lead to serious dermatological problems, which include melasma, melanoderma, lentigo, and vitiligo. Therefore, regulating melanin production is important to prevent the pigmentation disorders. Myanmar has a rich in natural resources. However, the chemical constituents of these natural resources in Myanmar have not been fully investigated. In the effort to search for compounds with anti-melanin deposition activity from Myanmar natural resources, five plants were collected in Myanmar. Extracts of these collected five plants were tested for anti-melanin deposition activity against a mouse melanoma cell line (B16-F10) induced with ${\alpha}$-melanocyte-stimulating hormone (${\alpha}$-MSH) and 3-isobutyl-1-methylxanthine (IBMX), and their anti-melanin deposition activities were compared with the positive control, arbutin. Among the tested extracts, the CHCl3 extracts of the Premna serratifolia (syn: P. integrifolia) wood showed anti-melanin deposition activities with IC50 values of $81.3{\mu}g/mL$. Hence, this study aims to identify secondary metabolites with anti-melanin deposition activity from P. serratifolia wood of Myanmar. P. serratifolia belongs to the Verbenaceae family and is widely distributed in near western sea coast from South Asia to South East Asia, which include India, Malaysia, Vietnam, Cambodia, and Sri Lanka. People in Tanintharyi region located in the southern part of Myanmar utilize the P. serratifolia, Sperethusa crenulata, Naringi crenulata, and Limonia acidissima as Thanaka, traditional cosmetics in Myanmar. Thanaka is applied in the form of paste onto skins to make it smooth and clear, as well as to prevent wrinkles, skin aging, excessive facial oil, pimples, blackheads, and whiteheads. However, the chemical constituents responsible for their cosmetic properties are yet to be identified. Moreover, the chemical constituents of P. serratifolia was almost uncharacterized. Investigation of the P. serratifolia chemical constituents is thus an attractive endeavor to discover new anti-melanin deposition active compounds. The investigation of the chemical constituents of the active CHCl3 extract of P. serratifolia led to isolation of four new lignoids, premnan A (1), premnan B (2), taungtangyiol C (3), and 7,9-dihydroxydolichanthin B (4), together with premnan C (5) (assumed to be an artifact), one natural newlignoid,(3R,4S)-4-(1,3-benzodioxol-5-ylcarbonyl)-3-[(R)-1-(1,3-benzo dioxol-5-yl)-1-hydroxy methyl]tetrahydro-2-furanone (6), and five known compounds (7-11)1,2). The structures of all isolated compounds were determined on the basis of their spectroscopic data and by comparison with the reported literatures. The absolute configurations of 1-3 and 5 were also determined by optical rotation and circular dichroism (CD) data analyses1). The anti-melanin deposition activities of all the isolated compounds were evaluated against B16-F10 cell line. 7,9-Dihydroxydolichanthin B (4) and ($2{\alpha},3{\alpha}$)-olean-12-en-28-oic acid (11) showed strong anti-melanin deposition activities with IC50 values of 18.4 and $11.2{\mu}M$, respectively, without cytotoxicity2). On the other hand, compounds 1-3, 5, and 7 showed melanogenesis enhancing activities1). To better understand their anti-melanin deposition mechanism, the effects of 4 and 11 on tyrosinase activities were investigated. The assay indicated that compounds 4 and 11 did not inhibit tyrosinase. Furthermore, we also examined the mRNA expression of microphthalmia-associated transcription factor (MITF), tyrosinase (TYR), tyrosinase-related protein-1 (TRP-1), and tyrosinase-related protein-2 (TRP-2). Compounds 4 and 11 down-regulated the expression of Tyr and Mitf mRNAs, respectively. Although the P. serratifolia wood has been used as traditional cosmetics in Myanmar for centuries, there are no scientific evidences to support its effectiveness as cosmetics. Investigation of the anti-melanin deposition activity of the chemical constituents of P. serratifolia thus provided insight into the effectiveness of the P. serratifolia wood as a cosmetic agent.

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