• Title/Summary/Keyword: Mouse skin

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In vitro Anti-allergic Effects of Bangpungtongseong-san in Human Keratinocytes and Primary Mouse Splenocytes (간 각질 세포 및 마우스 비장 세포를 이용한 방풍통성산의 항알러지 효능 연구)

  • Jeong, Soo-Jin;Lee, Mee-Young;Seo, Chang-Seob;Shin, Hyeun-Kyoo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.29 no.2
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    • pp.168-173
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    • 2015
  • Bangpungtongseong-san(BPTSS, Fangfengtongsheng-san in Chineses) is a traditional herbal formula comprising 18 medicinal herbs. In the present study, we performed the simultaneous analysis for four compounds of BPTSS and examined anti-allergic effects in human keratinocytes and mouse splenocytes. The column for separation of four compounds was used Luna C18 column and maintained at 40℃. The mobile phase for gradient elution consisted of two solvent systems. The analysis was carried out at a flow rate of 1.0 mL/min with PDA detection at 254 and 280 nm. To evaluate production and expression of Th2 chemokines, ELISA and RT-PCR were conducted in tumor necrosis factor (TNF)-α and interferon (IFN)-γ-stimulated HaCaT cells with or without BPTSS or silymarin, a positive control for skin inflammation. To measure Th2 cytokines, primary mouse splenocytes were treated with BPTSS and performed ELISA for interleukin (IL)-4, 5, 13. Calibration curves were acquired with r2>0.9999. The contents of geniposide, liquiritin, baicalin, and glycyrrhizin in BPTSS were 5.06 ㎎/g, 7.33 ㎎/g, 27.56 ㎎/g, and 7.81 ㎎/g, respectively. BPTSS reduced TARC and RANTES production and mRNA expression in TNF-α and IFN-γ-treated HaCaT cells. BPTSS inhibited IL-4, 5, and 13 production in mouse splenocytes. Our data will be a helpful information to upgrade quality control and anti-allergic effects of BPTSS.

Effect of Sopung-san on Contact Hypersensitivity induced by Repeat Elicitation of DNCB (소풍산물추출물이 DNCB로 유발된 접촉성피부염에 미치는 영향)

  • Park, Sook-Jahr;Lee, Sang-Gon;Baek, Jung-Han;Byun, Sung-Hui;Kim, Sang-Chan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.3
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    • pp.623-628
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    • 2006
  • Sopung-san(SPS; prescription No. DO27), a herbal medicine prescribed by oriental hospital of Daegu fanny University, has been used as cure for pruritus and psoriasis. To evaluate anti-inflammatory effect of Sopung-san, we treated Sopung-san extract in animal model system induced contact hypersensitivity. Contact hypersensitivity, a local inflammatory response of the skin, was induced by 1% DNCB on the right ear of BALB/C mouse. Sopung-san was prepared as water extract and administrated everyday for 2 weeks per oral. A right ear of mouse was potently swelled by 1% of DNCB treatment, but a mouse ear thickness was significantly reduced by Sopung-san after 2 weeks. treatment. Sopung-san reduced IgG in serum obtained from blood of 1 % DNCB-treated mouse. IgE in serum was not changed by DNCB and/or Sopung-san treatment. $IL-1{\beta}$ and IL-6 from ear tissue were significantly increased by DNCB, Dut were not reduced by Sopung-san. From these results, anti-inflammatory effect of Sopung-san, especially reduction of ear swelling, might be partly due to reduction of IgG in serum.

Effect of Eucommia ulmoides Extracts on Allergic Contact Dermatitis and Oxidative Damage Induced by Repeat Elicitation of DNCB (두충 추출물이 DNCB로 유발된 알레르기성 접촉피부염과 산화적 손상에 미치는 영향)

  • Shon, Mi-Yae;Nam, Sang-Hae
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.12
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    • pp.1517-1522
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    • 2007
  • Inhibitory effects of allergic contact dermatitis of hot water extract of Duchung (Eucommia ulmoides Oliver) leaf, bark and stem growing at Sancheong-gun were investigated for female BALB/c mouse induced by repeat elicitation of DNCB (2,4-dinitro-chlorobenzene). Skin reactions, consisting of increased ear thickness and the presence of ear inflammation, were observed in mice treated with DNCB and Duchung. Weight of lymph node, spleen and thymus in mice treated with Duchung extracts were lower than that of mouse treated with DNCB. Ear weight of mouse treated with Duchung extracts was decreased by increasing the concentration of sample as compared to control group and dropped as low as control level at 1,000 mg/kg. Ear thickness became thinner as test time on Duchung extract progressed. MDA (malondialdehyde) contents in liver tissue were not different in sample group with DNCB group, but were different in ear tissue. NO (nitric oxide) contents was decreased in Duchung extract groups at serum and ear tissues as compared to 1% DNCB group. In the present study, the results suggested that Duchung extract inhibits inflammatory response and oxidative damage induced DNCB allergen.

The Genome-Wide Expression Profile of Saussurea lappa Extract on House Dust Mite-Induced Atopic Dermatitis in Nc/Nga Mice

  • Lim, Hye-Sun;Ha, Hyekyung;Shin, Hyeun-Kyoo;Jeong, Soo-Jin
    • Molecules and Cells
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    • v.38 no.9
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    • pp.765-772
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    • 2015
  • Saussurea lappa has been reported to possess anti-atopic properties. In this study, we have confirmed the S. lappa's anti-atopic properties in Nc/Nga mice and investigated the candidate gene related with its properties using microarray. We determined the target gene using real time PCR in in vitro experiment. S. lappa showed the significant reduction in atoptic dermatitis (AD) score and immunoglobulin E compared with the AD induced Nc/Nga mice. In the results of microarray using back skin obtained from animals, we found that S. lappa's properties are closely associated with cytokine-cytokine receptor interaction and the JAK-STAT signaling pathway. Consistent with the microarray data, real-time RT-PCR confirmed these modulation at the mRNA level in skin tissues from S. lappa-treated mice. Among these genes, PI3Kca and $IL20R{\beta}$ were significantly downregulated by S. lappa treatment in Nc/Nga mouse model. In in vitro experiment using HaCaT cells, we found that the S. lappa components, including alantolactone, caryophyllene, costic acid, costunolide and dehydrocostus lactone significantly decreased the expression of PI3Kca but not $IL20R{\beta}$ in vitro. Therefore, our study suggests that PI3Kca-related signaling is closely related with the protective effects of S. lappa against the development of atopic-dermatitis.

Isolation from Gloydius blomhoffii siniticus Venom of a Fibrin(ogen)olytic Enzyme Consisting of Two Heterogenous Polypeptides

  • Choi, Suk-Ho;Lee, Seung-Bae
    • Journal of Pharmacopuncture
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    • v.16 no.2
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    • pp.46-54
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    • 2013
  • Objective: This study was undertaken to isolate a fibrin(ogen)olytic enzyme from the snake venom of Gloydius blomhoffii siniticus and to investigate the enzymatic characteristics and hemorrhagic activity of the isolated enzyme as a potential pharmacopuncture agent. Methods: The fibrinolytic enzyme was isolated by using chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and fibrin plate assay. The characteristics of the enzyme were determined by using fibrin plate assay, protein hydrolysis analysis, and hemorrhage assay. Its amino acid composition was determined. Results: The fibrin(ogen)olytic enzyme with the molecular weight of 27 kDa (FE-27kDa) isolated from G. b. siniticus venom consisted of two heterogenous disulfide bond-linked polypeptides with the molecular weights of 15 kDa and 18 kDa. When more than $20{\mu}g$ of FE-27kDa was applied on the fibrin plate, fibrinolysis zone was formed as indicating its fibrinolytic activity. The fibrinolytic activity was inhibited completely by phenylmethanesulfonylfluoride (PMSF) and ethylenediaminetetraacetic acid (EDTA) and partially by thiothreitol and cysteine. Metal ions such as $Hg^{2+}$ and $Fe^{2+}$ inhibited the fibrinolytic activity completely, but $Mn^{2+}$ did not. FE-27kDa preferentially hydrolyzed ${\alpha}$-chain of fibrinogen and slowly hydrolyzed ${\beta}$-chain, but did not hydrolyze ${\gamma}$-chain. High-molecular-weight polypeptides of gelatin were hydrolyzed partially into polypeptides with molecular weights of more than 45 kDa. A dosage of more than $10{\mu}g$ of FE-27kDa per mouse was required to induce hemorrhage beneath the skin. Conclusion: FE-27kDa was a serine proteinase consisting of two heterogeneous polypeptides, hydrolyzed fibrin, fibrinogen, and gelatin, and caused hemorrhage beneath the skin of mouse. This study suggests that the potential of FE-27kDa as pharmacopuncture agent should be limited due to low fibrinolytic activity and a possible side effect of hemorrhage.

External Application of Apo-9'-fucoxanthinone, Isolated from Sargassum muticum, Suppresses Inflammatory Responses in a Mouse Model of Atopic Dermatitis

  • Han, Sang-Chul;Kang, Na-Jin;Yoon, Weon-Jong;Kim, Sejin;Na, Min-Chull;Koh, Young-Sang;Hyun, Jin-Won;Lee, Nam-Ho;Ko, Mi-Hee;Kang, Hee-Kyoung;Yoo, Eun-Sook
    • Toxicological Research
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    • v.32 no.2
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    • pp.109-114
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    • 2016
  • Allergic skin inflammation such as atopic dermatitis is characterized by skin barrier dysfunction, edema, and infiltration with various inflammatory cells. The anti-inflammatory effects of Apo-9'-fucoxanthinone, isolated from Sargassum muticum, have been described in many diseases, but the mechanism by which it modulates the immune system is poorly understood. In this study, the ability of Apo-9'-fucoxanthinone to suppress allergic reactions was investigated using a mouse model of atopic dermatitis. The Apo-9'-fucoxanthinone-treated group showed significantly decreased immunoglobulin E in serum. Also, Apo-9'-fucoxanthinone treatment resulted in a smaller lymph node size with reduced the thickness and length compared to the induction group. In addition, Apo-9'-fucoxanthinone inhibited the expression of interleukin-4, interferon-gamma and tumor necrosis factor-alpha by phorbol 12-myristate 13-acetate and ionomycin-stimulated lymphocytes. These results suggest that Apo-9'-fucoxanthinone may be a useful therapeutic strategy for treating chronic inflammatory diseases.

Formulation Design and Evaluation of Ketorolac Tromethamine Hydrogel for Transdermal Delivery System (경피흡수를 위한 케토롤락 하이드로겔의 제제설계 및 평가)

  • Cho, In-Sook;Lee, Gye-Won;Lee, Jong-Hwa;Jee, Ung-Kil
    • Journal of Pharmaceutical Investigation
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    • v.33 no.1
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    • pp.21-28
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    • 2003
  • Ketorolac tromethamine(KT) is a nonsteroidal agent with potent analgesic and moderate anti-inflammatory activity. The lipid-water partition coefficient of KT was evaluated and KT gel was formulated as a gel containing different pH, different concentrations of polymer (poloxamer 407, carbopol 941), propylene glycol, ethanol and various enhancers. The resulting KT gels were evaluated with respect to their viscosity, in vitro drug permeation rate through hairless mouse skin and stability. In n-octanol and chloroform, the lipid-water partition coefficient of KT was the highest at pH 4 phosphate buffer. The apparent viscosity of KT gel increased with an increase in gel pH, polymer and enhancer concentration. But the apparent viscosity of KT gel decreased with an increase in ethanol concentration. The permeation rate of KT through hairless mouse skin from gels different pH was maximum at pH 4 which is close to KT $pK_{a}$ 3.54. The permeation rate decreased with an increase in polymer, propylene glycol concentration. But the permeation rate increased with an increase in ethanol. The increase of drug concentration from 1 to 3% induced linear increase in permeation rate. The best enhancer was the combination of $Labrasol^{\circledR},\;Transcutol^{\circledR}$, oleic acid and l-menthol. In the accelerated stability test(25, 40 and $50{\circ}C$), pH 5 gel was most stable and pH 4 gel was most unstable for 90 days.

The Inhibitory Effect of Premature Citrus unshiu Extract on Atopic Dermatitis In Vitro and In Vivo

  • Kang, Gyeoung-Jin;Han, Sang-Chul;Yi, Eun-Jou;Kang, Hee-Kyoung;Yoo, Eun-Sook
    • Toxicological Research
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    • v.27 no.3
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    • pp.173-180
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    • 2011
  • Atopic dermatitis (AD) is a chronic, recurrent inflammatory skin disease that is associated with Th2 cell-mediated allergy. The process that leads to infiltration of inflammatory cells into an AD lesion is remarkably dependent on various chemokines, especially TARC (thymus and activation-regulated chemokine/CCL17) and MDC (macrophage-derived chemokine/CCL22). Serum levels of these chemokines are over-expressed in AD patients. Citrus unshiu, which is known as Satsuma mandarin, has anti-oxidative, anti-inflammation, and anti-microviral activity. Therefore, we investigated the effect of EtOH extract of premature C. unshiu on AD. We did this using a DNCB-induced AD mouse model. We also tried to confirm an inhibitory effect for premature C. unshiu on the expression of inflammatory chemokines in IFN-${\gamma}$ and TNF-${\alpha}$ stimulated HaCaT human keratinocytes. We found that extract of premature C. unshiu reduced DNCB-induced symptoms such as hyperkeratosis, increased skin thickness, and infiltrated mast cells, in our AD-like animal model. The extract decreased levels of IFN-${\gamma}$ and IL-4 in ConA-stimulated splenocytes isolated from DNCB-treated mice. Also, extract of premature C. unshiu inhibited mRNA expression and protein production of TARC and MDC through the inhibition of STAT1 phosphorylation. These results suggest that C. unshiu has anti-atopic activity by regulating inflammatory chemokines such as TARC and MDC.

The Effect of Low-power Laser on the Murine Hair Growth (저출력 레이저가 마우스 모발성장에 미치는 영향)

  • Chung, Phil Sang;Kim, Jin Wang;Lee, Jeong Ok;Ree, Chung Ku;Oh, Chung Hoon;Kim, You Chan;Chung, Sang Woon
    • Archives of Plastic Surgery
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    • v.32 no.2
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    • pp.149-154
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    • 2005
  • Low-power laser(LPL) delivers a small amount of energy without elevation of tissue temperature. LPL has been reported to have biostimulation effects including anti-inflammatory, analgesic, regenerative, immunocorrective, and vasodilative effects. However, the effect of LPL on hair growth has rarely been studied. We investigated the effect of LPL on hair growth in the mouse. After depilation of back skin of mice, we classified the mice into 4 groups: control, laser irradiated group, $MoandMore^{(R)}$ applied group, and Spella $707^{(R)}$ applied group. Laser irradiation or application of these drugs were performed on the back skin of the mice for 30 days. The results are summarized as follows. Hair growth of control was first observed at 13 days after depilation, and complete hair regrowth was observed at 25 days. Hair growth of both laser irradiation group and $MoandMore^{(R)}$ applied group was first observed at 9 days after depilation, and complete hair regrowth was observed at 20 days. Hair growth of Spella $707^{(R)}$ applied group was firstly observed at the 9 days after depilation, and complete hair regrowth was observed at the 15 days. Hair growth started at the irradiation site in the laser irradiation group, but it started at the random sites in other groups. In conclusion LPL irradiation have a stimulating effect on the hair growth in the mouse.

Transdermal Drug Delivery & Therapeutic Effect of the Preparations of Lithospermi Radix and Gardeniae Fructus Extracts on the Burn & Wound Healing (화상 및 창상에 대한 자근.치자 복합제제의 경피 흡수 및 치료효과)

  • Min, Dong-Hoon;Kim, Dae-Keun;Lim, Jong-Pil;Yang, Jae-Heon
    • Journal of Pharmaceutical Investigation
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    • v.35 no.4
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    • pp.255-263
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    • 2005
  • Research was undertaken to compare the pharmacological activity of Lithospermi radix (LR) reported as an oriental medicine for classical uses. LR contains naphthoquinone pigments : shikonin, acetylshikonin, isobutylshikonin, etc. LR is used for the treatment of excision wound, burn, eczema, blister, scarlatina and septicemia as antifebrile, antidotic and antiphlogistic. Gardeniae fructus (GF) has been used for the treatment to jaundice, hepatic disease, anti-inflammatory and analgesic effects, and it contains crocin, geniposide and its derivatives. The therapeutic effects of burn and excision wound healing from LR & GF hydrogel with $Nano-ATP^{\circledR}$ (GLN) were investigated. To evaluate the therapeutic value of various hydrogels, thermal burn model and excision wound mouse model were used. The burn and wound reduction rate and therapeutic period were measured to calculate the healing extent after 5 experiments. The 2nd degree burn was prepared on hairless mouse back skin and dressing with collagen. The burn and wound reduction rate of GLN hydrogel treated group decreased more rapidly than that of other gel group in animal model. Furthermore therapeutic periods of GLN hydrogel treated group was shorter than that of other gel group. In anti-inflammatory test, GLN hydrogel treated group decreased edema rapidly than that of other gel group. These results suggest that the GLN hydrogel treatment has an therapeutic effect on burn and excision wound healing.