• 대한임상검사과학회지
• /
• 제48권3호
• /
• pp.188-195
• /
• 2016

본 연구에서는 집먼지 진드기 추출물은 호중구에 단독으로 작용하는 것보다, 림프구와 호중구의 공동배양에서 호중구의 세포고사를 더 억제시켰다. 집먼지 진드기는 알레르기 질환의 림프구에서 IL-6, IL-8, MCP-1, GM-CSF의 분비를 증가시켰다. 집먼지 진드기에 의해 증가된 사이토카인은 protein kinase C ${\delta}$의 억제제인 rottlerin과 p38 MAPK의 억제제인 SB202190에 의해서 감소하였다. 집먼지 진드기에 의해 활성화된 p38 MAPK은 protease-activated receptor (PAR2)의 억제제, rottlerin, SB202190에 의해서 억제되었다. Serine protease 억제제인 aprotinin과 cysteine protease 억제제인 E64은 림프구의 사이토카인의 증가와 관련이 없었다. 또한 집먼지 진드기에 의해 증가된 사이토카인의 변화는 천식과 알레르기 비염 환자에서 차이가 없었다. 림프구에서 집먼지진드기에 의해서 분비되는 분자들은 호중구의 유주운동을 억제시켰다. 본 연구를 통하여 집먼지진드기에 의해 유발되는 알레르기 질환의 병인기전을 규명하는데 유용한 결과가 될 것이다.

• Biomolecules & Therapeutics
• /
• 제20권5호
• /
• pp.463-469
• /
• 2012

Atopic dermatitis is a chronic, inflammatory disease of the skin with increased transepidermal water loss. Both an abnormal inflammatory response and a defective skin barrier are known to be involved in the pathogenesis of atopic dermatitis. Protease activated receptor 2 (PAR2) belongs to a family of G-protein coupled receptors and is activated by both trypsin and a specific agonist peptide, SLIGKV-$NH_2$. PAR2 is expressed in suprabasal layers of the epidermis and regulates inflammatory responses and barrier homeostasis. In this study, we show that nordihydroguaiaretic acid (NDGA) inhibits the PAR2-mediated signal pathway and plays a role in skin barrier recovery in atopic dermatitis. Specifically, NDGA reduces the mobilization of intracellular $Ca^{2+}$ in HaCaT keratinocytes by down-regulating inflammatory mediators, such as interleukin-8, thymus and activation-regulated chemokine and intercellular cell adhesion molecule-1 in HaCaT keratinocytes. Also, NDGA decreases the protein expression of involucrin, a differentiation maker of keratinocyte, in both HaCaT keratinocytes and normal human epidermal keratinocytes. We examined NDGA-recovered skin barrier in atopic dermatitis by using an oxazolone-induced atopic dermatitis model in hairless mice. Topical application of NDGA produced an increase in transepidermal water loss recovery and a decrease in serum IgE level, without weight loss. Accordingly, we suggest that NDGA acts as a PAR2 antagonist and may be a possible therapeutic agent for atopic dermatitis.

• The Korean Journal of Physiology and Pharmacology
• /
• 제25권1호
• /
• pp.15-26
• /
• 2021

Peptides are short chain of amino acids linked by peptide bonds. They are widely used as effective and biocompatible active ingredients in cosmetic industry. In this study, we developed novel peptide mixture and identified its anti-pigmentation effect on melanocytes and keratinocytes. Our results revealed that peptide mixture inhibited melanosome biogenesis through the regulation of microphthalmia-associated transcription factor, a key factor of melanogenesis in melanocytes. And we observed that peptide mixture inhibited melanosome uptake through the reduction of protease-activated receptor 2, a phagocytosis-related receptor in keratinocytes. Furthermore, peptide mixture activated autophagy system resulting in degradation of transferred melanosomes in keratinocytes. The anti-pigmentation effect of multi-targeting peptide mixture was assessed in a human skin equivalent model (MelanoDerm). Melanin contents in epidermal layer were significantly decreased by topical treatment of peptide mixture, suggesting that it can be applied as a novel cosmetics material having a whitening function.

• Asian Pacific Journal of Cancer Prevention
• /
• 제15권2호
• /
• pp.643-646
• /
• 2014

Objective: This study aimed to explore the expression of tissue factor (TF), protease activated receptor-2 (PAR-2), and matrix metalloproteinase-9 (MMP-9) in the MCF-7 breast cancer cell line and influence on invasiveness. Methods: Stable MCF-7 cells transfected with TF cDNA and with TF ShRNA were established. TF, PAR-2, and MMP-9 protein expression was analyzed using indirect immunofluorescence and invasiveness was evaluated using a cell invasion test. Effects of an exogenous PAR-2 agonist were also examined. Results: TF protein expression significantly differed between the TF cDNA and TF ShRNA groups. MMP-9 protein expression was significantly correlated with TF protein expression, but PAR-2 protein expression was unaffected. The PAR-2 agonist significantly enhanced MMP-9 expression and slightly increased TF and PAR-2 expression in the TF ShRNA group, but did not significantly affect protein expression in MCF-7 cells transfected with TF cDNA. TF and MMP-9 expression was positively correlated with the invasiveness of tumor cells. Conclusion: TF, PAR-2, and MMP-9 affect invasiveness of MCF-7 cells. TF may increase MMP-9 expression by activating PAR-2.

• Biomolecules & Therapeutics
• /
• 제31권1호
• /
• pp.59-67
• /
• 2023

Thrombin is a serine protease that participates in a variety of biological signaling through protease-activated receptors. Intestinal myofibroblasts play central roles in maintaining intestinal homeostasis. In this study, we found that thrombin-induced apoptosis is mediated by the calcium-mediated activation of cytosolic phospholipase A₂ in the CCD-18Co cell. Thrombin reduced cell viability by inducing apoptosis and proteinase-activated receptor-1 antagonist attenuated thrombin-induced cell death. Endogenous ceramide did not affect the cell viability itself, but a ceramide-mediated pathway was involved in thrombin-induced cell death. Thrombin increased intracellular calcium levels and cytosolic phospholipase A₂ activity. The ceramide synthase inhibitor Fumonisin B1, intracellular calcium chelator BAPTA-AM, and cytosolic phospholipase A₂ inhibitor AACOCF₃ inhibited thrombin-induced cell death. Thrombin stimulated arachidonic acid release and reactive oxygen species generation, which was blocked by AACOCF₃, BAPTA-AM, and the antioxidant reagent Trolox. Taken together, thrombin triggered apoptosis through calcium-mediated activation of cytosolic phospholipase A₂ in intestinal myofibroblasts.

• Biomolecules & Therapeutics
• /
• 제24권5호
• /
• pp.529-535
• /
• 2016

Atopic dermatitis (AD) results from gene and environment interactions that lead to a range of immunological abnormalities and breakdown of the skin barrier. Protease-activated receptor 2 (PAR2) belongs to a family of G-protein coupled receptors and is expressed in suprabasal layers of the epidermis. PAR2 is activated by both trypsin and a specific agonist peptide, SLIGKV-$NH_2$ and is involved in both epidermal permeability barrier homeostasis and epithelial inflammation. In this study, we investigated the effect of lobaric acid on inflammation, keratinocyte differentiation, and recovery of the skin barrier in hairless mice. Lobaric acid blocked trypsin-induced and SLIGKV-$NH_2$-induced PAR2 activation resulting in decreased mobilization of intracellular $Ca^{2+}$ in HaCaT keratinocytes. Lobaric acid reduced expression of interleukin-8 induced by SLIGKV-$NH_2$ and thymus and activation regulated chemokine (TARC) induced by tumor necrosis factor-a (TNF-${\alpha}$) and IFN-${\gamma}$ in HaCaT keratinocytes. Lobaric acid also blocked SLIGKV-$NH_2$-induced activation of ERK, which is a downstream signal of PAR2 in normal human keratinocytes (NHEKs). Treatment with SLIGKV-$NH_2$ downregulated expression of involucrin, a differentiation marker protein in HaCaT keratinocytes, and upregulated expression of involucrin, transglutamase1 and filaggrin in NHEKs. However, lobaric acid antagonized the effect of SLIGKV-$NH_2$ in HaCaT keratinocytes and NHEKs. Topical application of lobaric acid accelerated barrier recovery kinetics in a SKH-1 hairless mouse model. These results suggested that lobaric acid is a PAR2 antagonist and could be a possible therapeutic agent for atopic dermatitis.

• 대한한방소아과학회지
• /
• 제35권3호
• /
• pp.128-137
• /
• 2021

Objective The purpose of this study was to confirm the effectiveness of coptidis rhizoma extract-ceramide complex on skin barrier, transepidermal water loss (TEWL) and pH reduction, and inflammation of the skin. Methods Coptidis rhizoma extract-ceramide complex was applied in 6-week-old Balb/C mice after dermatitis was induced. To confirm the skin condition changes, TEWL and pH were observed, and filaggrin in the stratum corneum of the skin was observed. Kallikrein-related peptidase (KLK) 7, Protease activated receptor (PAR)-2, Thymic stromal lymphopoietin (TSLP), and IL-4 were observed in the stratum corneum to confirm the changes in the inflammatory response. Results Filaggrin positive reaction was increased in the experiment group compared to the control group. TEWL and pH were lower in the experiment group compared to the control group. KLK7, PAR2, TSLP, and IL-4 positive responses were decreased in the experiment group compared to the control group. Conclusions It was confirmed that the coptidis rhizoma extract-ceramide complex can relieve the inflammatory response of atopic dermatitis by restoring the skin lipid barrier damage.

• Asian Pacific Journal of Cancer Prevention
• /
• 제14권6호
• /
• pp.3793-3798
• /
• 2013

The role of protease-activated receptors (PARs) in lung tumors is controversial. Although PAR4 is preferentially expressed in human lung tissues, its possible significance in lung cancer has not been defined. The studies reported herein used a combination of clinical observations and molecular methods. Surgically resected lung adenocarcinomas and associated adjacent normal lung tissues were collected and BEAS-2B and NCI-H157 cell lines were grown in tissue culture. PAR4 expression was evaluated by RT-PCR, RT-qPCR, Western blotting and immunohistochemistry analysis. The results showed that PAR4 mRNA expression was generally decreased in lung adenocarcinoma tissues as compared with matched noncancerous tissues (67.7%) and was associated with poor differentiation (p=0.017) and metastasis (p=0.04). Western blotting and immunohistochemical analysis also showed that PAR4 protein levels were mostly decreased in lung adenocarcinoma tissues (61.3%), and were also associated with poor differentiation (p=0.035) and clinical stage (p=0.027). Moreover, PAR4 expression was decreased in NCI-H157 cells as compared with BEAS-2B cells. In conclusion, PAR4 expression is significantly decreased in lung adenocarcinoma, and down-regulation of PAR4 is associated with a more clinically aggressive phenotype. PAR4 may acts as a tumor suppressor in lung adenocarcinoma.

• 대한한방소아과학회지
• /
• 제35권4호
• /
• pp.156-166
• /
• 2021

Objectives The purpose of this study is to confirm the regulate effect of T helper (Th) 2 differentiation that Baekho-tang extract may produce to improves skin lipid barrier damages. Methods Four-weeks-old NC/Nga mice were divided into four groups: control group (Ctrl, n=10), lipid barrier eliminated group (LBE, n=10), Dexamethasone treatment after lipid barrier elimination group (DxT, n=10), and Baekho-tang extract treatment group after lipid barrier elimination group (BHTT, n=10). Baeko-tang extract was administered for 3 days after removal of the skin fat barrier in BHTT group. Then, we identified changes in external symptoms of the skin, factors affecting skin barrier such as potential of hydrogen (pH), filaggrin (FLG), transepidermal water loss (TEWL) and Th2 differentiation factors like Interleukin (IL)-4, Kallikrein Related Peptidase 7 (KLK7) and protease activated receptor 2 (PAR-2) through our immunohistochemistry. Results After lipid barrier elimination, the reduction of morphological skin inflammations was less in BHTT group than in LBE group and DxT group. Also, pH and TEWL were significantly decreased with BHTT group. However, FLG was significantly increased in BHTT group compared to LBE, DxT, and Ctrl group. All kinds of Th2 differentiation factors (IL-4, KLK7 and PAR-2) were also decreased in BHTT compared to the LBE and DxT. Conclusions As a result of this study, BHT administration decreased pH, TEWL, and increased FLG, thus participating in recovering damaged skin barrier. Since Th2 differentiation factors were decreased as well, BHT's regulatory effect in sequential immune reactions may be a possible explanation of how it enhances recovery of the damaged lipid barrier.

• 대한한방소아과학회지
• /
• 제36권1호
• /
• pp.57-64
• /
• 2022

Objectives The purpose of this study is to confirm the regulate effect of T helper (Th) 2 differentiation that Sabaek-san extract may produce to improves skin lipid barrier damage. Methods Four-weeks-old NC/Nga mice were divided into four groups: control group (Ctrl, n=10), lipid barrier eliminated group (LBE, n=10), Dexamethasone treatment after lipid barrier elimination group (DXT, n=10), and Sabaek-san extract treatment group after lipid barrier elimination (SBT, n=10). Sabaek-san extract was administered for 3 d after removal of the skin fat barrier in SBT group. Then, we identified changes in external symptoms of the skin, factors affecting skin barrier such as potential of hydrogen (pH), filaggrin, transepidermal water loss (TEWL) and Th2 differentiation factors like Interleukin (IL)-4, Kallikrein Related Peptidase 7 (KLK7), and protease activated receptor 2 (PAR-2) through our immunohistochemistry. Results After lipid barrier elimination, the reduction of morphological skin inflammations was less in SBT group than in LBE and DXT group. Also, pH and TEWL were significantly decreased with SBT group. However, filaggrin was significantly increased in SBT group compared to LBE, DXT, and Ctrl group. All kinds of Th2 differentiation factors (IL-4, KLK7 and PAR-2) were also decreased in SBT compared to the LBE and DXT. Conclusions As a result of this study, SBT administration decreased pH, TEWL, and increased filaggrin, thus participating in recovering damaged skin barrier. Since Th2 differentiation factors were decreased as well, SBT's regulatory effect in sequential immune reactions may be a possible explanation of how it enhances recovery of the damaged lipid barrier.