Gasdermin C is induced by ultraviolet light and contributes to MMP-1 expression via activation of ERK and JNK pathways

Kusumaningrum, Novi,Lee, Dong Hun,Yoon, Hyun-Sun,Kim, Yeon Kyung,Park, Chi-Hyun,Chung, Jin Ho Elsevier 2018 Journal of dermatological science Vol.90 No.2

<P><B>Abstract</B></P> <P><B>Background</B></P> <P>Ultraviolet (UV) radiation plays important roles in various skin diseases including premature aging and cancer. UV has been shown to regulate the expressions of many genes including matrix metalloproteinases (MMPs). Gasdermin C (GSDMC) belongs to Gasdermin family and is known to be expressed in the epithelial cells of many tissues including the skin. However, the functions of GSDMC remain poorly understood.</P> <P><B>Objective</B></P> <P>We aimed to investigate the role of GSDMC in UV-induced MMP-1, MMP-3, and MMP-9 expressions in human skin keratinocytes.</P> <P><B>Methods</B></P> <P>Primary human skin keratinocytes and an immortalized human skin keratinocyte cell line (HaCaT cells) were irradiated with UV. Knockdown and overexpression of GSDMC were performed to study the effect of GSDMC. The mRNA and protein levels were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting, respectively.</P> <P><B>Results</B></P> <P>We found that GSDMC expression is increased by UV irradiation in human skin keratinocytes. Further studies showed that GSDMC expression is increased at relatively late time points after UV irradiation and that this GSDMC induction plays important roles in the expressions of MMP-1, but not of MMP-3 and MMP-9, and the activations of ERK and JNK induced by UV. In addition, we found that overexpression of GSDMC increases the MMP-1 expression and the activities of ERK and JNK and that GSDMC-induced MMP-1 expression is suppressed by inhibition of ERK or JNK activities.</P> <P><B>Conclusions</B></P> <P>Our results suggest that GSDMC is increased by UV radiation and contributes to UV-induced MMP-1 expression through the activation of ERK and JNK pathways.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Expression of GSDMC is increased by UV irradiation in human skin keratinocytes. </LI> <LI> Increase of GSDMC by UV contributes to the expression of MMP-1 and the activation of ERK and JNK induced by UV. </LI> <LI> GSDMC plays an important role in UV-induced MMP-1 expression through the activation of ERK and JNK pathways in human skin keratinocytes. </LI> </UL> </P>

Protective Effects of Portulaca oleracea L. Extract against Matrix Metalloproteinase Production and Reactive Oxygen Species Generation Induced by Ultraviolet B Radiation in Human Keratinocytes

Jung Hwan Oh(오정환),Fatih Karadeniz(파티 카라데니즈),Jung Im Lee(이정임),So Young Park(박소영),Youngwan Seo(서영완),Chang-Suk Kong(공창숙) 한국생명과학회 2018 생명과학회지 Vol.28 No.8

쇠비름(Portulaca oleracea.L)은 쇠비름과에 속하는 한해살이풀로서 리놀렌산과 같은 불포화지방산, 페놀성 화합물, 플라보노이드, 비타민 C, 미네랄 함량이 높은 것으로 보고되어 있다. 본 연구에서는 쇠비름 추출물을 이용하여 UVB를 조사한 인간각질형성세포에서 광노화 억제능을 확인하였다. Matrix metalloproteinases는 세포의 기질을 분해하는 효소로 MMP-1는 collagenase, MMP-2와 MMP-9는 gelatinases로 피부 진피층을 구성하는 type Ⅰ collagen을 분해시키는데 영향을 미친다. UVB를 조사한 인간각질형성세포에서 쇠비름 추출물을 처리했을 때 MMP-1, -2, -9의 발현이 감소하였으며, type Ⅰ procollagen의 발현은 증가하는 것으로 나타났다. 또한 쇠비름 추출물을 처리한 군에서 UV에 의한 ROS 생성이 감소하였는데 이는 Nrf-2의 활성화를 통한 항산화 인자 SOD-1과 OH-1의 발현 증가로 인해 세포내 ROS 생성이 감소한 것으로 사료된다. 따라서 본 연구 결과를 통해 쇠비름 추출물이 UVB를 조사한 인간각질형성세포에서 MMP 인자 및 항산화 인자의 발현 조절을 통해 광노화로부터의 세포 보호능을 가지는 것을 확인하였으며 나아가 화장품 소재로서의 개발 가능성을 확인하였다. Portulaca oleracea L. is an edible plant widely consumed in daily diet throughout Europe, Asia and America. In this study, protective effects of P. oleracea L. extracts against oxidative stress and matrix metalloproteinase (MMP) activity induced by ultraviolet B (UVB) radiation were investigated using HaCaT immortal human keratinocytes. In this context, the mRNA and protein productions of MMPs (MMP-1, -2, and -9) and type I procollagen, which are major markers of photoaging induced by UVB radiation in HaCaT keratinocytes, were evaluated. Furthermore, UVB-induced reactive oxygen species (ROS) generation and mRNA and protein expression levels of superoxide dismutase-1 (SOD-1), oxygenase-1 (OH-1), and nuclear factor-erythroid 2-related factor-2 (Nrf-2), all of which are associated with the antioxidant balance, were investigated. As shown by the results, UVB radiation induced ROS formation and led to increased production of MMPs and decreased collagen production in human keratinocytes, which resulted in skin photoaging or photodamage. The treatment with P. oleracea L. extracts downregulated MMP (MMP-1, -2, and -9) production and upregulated type I procollagen expression in UVB-induced HaCaT cells. Furthermore, treatment with the extracts decreased UVB-induced ROS generation and increased the expression of antioxidant enzymes, such as SOD-1 and OH-1, through the Nrf-2 pathway. Taken together, these results suggest that P. oleracea L. extracts could be a potential cosmeceutical agent for the prevention of skin photoaging or photodamage.

In vitro evaluation method to monitor pathogenic sensitive skin by co-culture of keratinocytes and neuronal cell line

( Sun Mee Shin ),( Eun Hye Hong ),( Soo Hyun Jeong ),( Eun Joo Park ),( Kwang Joong Kim ),( Kwang Ho Kim ) 대한피부과학회 2020 대한피부과학회 학술발표대회집 Vol.72 No.1

Background: Keratinocytes have recently known to participate in sensory transduction by release of neuroactive molecules which bind to intra-epidermal free nerve endings (FNEs) and modulate nociception. Although reciprocal interactions between keratinocytes and FNEs via soluble mediators are well established, little is known about physical contacts between keratinocytes and sensory neurons. Objectives: We developed for the first time the co-culture of human primary keratinocyte with differentiated SH-SY5Y to reproduce direct interactions in vitro. Methods: We investigated the morphological and functional characteristics of differentiated SH-SY5Y neuronal cell line co-cultured with primary keratinocyte and analyzed the influence of keratinocytes. Results: SH-SY5Y cells survived well in keratinocyte co-culture condition. When SH-SY5Y cells were co-cultured with keratinocytes, they had no significant influence on axonal development. The neuropeptide Substance P (SP) which is released after a cytosolic calcium influx and modulates immediate skin hypersensitivity was also released well after capsaicin treatment in co-cultured condition. Conclusion: This co-culture model in which keratinocytes and neurons may be a useful in vitro alternative for studying and characterizing the close communication between keratinocytes and sensory neurons.

Diphlorethohydroxycarmalol Suppresses Ultraviolet B-Induced Matrix Metalloproteinases via Inhibition of JNK and ERK Signaling in Human Keratinocytes

( Mei Jing Piao ),( Madduma Hewage Susara Ruwan Kumara ),( Ki Cheon Kim ),( Kyoung Ah Kang ),( Hee Kyoung Kang ),( Nam Ho Lee ),( Jin Won Hyun ) 한국응용약물학회 2015 Biomolecules & Therapeutics(구 응용약물학회지) Vol.23 No.6

Skin aging is the most readily observable process involved in human aging. Ultraviolet B (UVB) radiation causes photo-oxidation via generation of reactive oxygen species (ROS), thereby damaging the nucleus and cytoplasm of skin cells and ultimately leading to cell death. Recent studies have shown that high levels of solar UVB irradiation induce the synthesis of matrix metalloproteinases (MMPs) in skin fibroblasts, causing photo-aging and tumor progression. The MMP family is involved in the breakdown of extracellular matrix in normal physiological processes such as embryonic development, reproduction, and tissue remodeling, as well as in disease processes such as arthritis and metastasis. We investigated the effect of diphlorethohydroxycarmalol (DPHC) against damage induced by UVB radiation in human skin keratinocytes. In UVB-irradiated cells, DPHC significantly reduced expression of MMP mRNA and protein, as well as activation of MMPs. Furthermore, DPHC reduced phosphorylation of ERK and JNK, which act upstream of c-Fos and c-Jun, respectively; consequently, DPHC inhibited the expression of c-Fos and c-Jun, which are key components of activator protein-1 (AP-1, up-regulator of MMPs). Additionally, DPHC abolished the DNA-binding activity of AP-1, and thereby prevented AP-1-mediated transcriptional activation. These data demonstrate that by inactivating ERK and JNK, DPHC inhibits induction of MMPs triggered by UVB radiation.

Isorhamnetin Protects Human Keratinocytes against Ultraviolet B-Induced Cell Damage

( Xia Han ),( Mei Jing Piao ),( Ki Cheon Kim ),( Susara Ruwan Kumara Madduma Hewage ),( Eun Sook Yoo ),( Young Sang Koh ),( Hee Kyoung Kang ),( Jennifer H Shin ),( Yeun Soo Park ),( Suk Jae Yoo ),( Su 한국응용약물학회 2015 Biomolecules & Therapeutics(구 응용약물학회지) Vol.23 No.4

Isorhamnetin (3-methylquercetin) is a flavonoid derived from the fruits of certain medicinal plants. This study investigated the photoprotective properties of isorhamnetin against cell damage and apoptosis resulting from excessive ultraviolet (UV) B exposure in human HaCaT keratinocytes. Isorhamnetin eliminated UVB-induced intracellular reactive oxygen species (ROS) and attenuated the oxidative modification of DNA, lipids, and proteins in response to UVB radiation. Moreover, isorhamnetin repressed UVB-facilitated programmed cell death in the keratinocytes, as evidenced by a reduction in apoptotic body formation, and nuclear fragmentation. Additionally, isorhamnetin suppressed the ability of UVB light to trigger mitochondrial dysfunction. Taken together, these results indicate that isorhamnetin has the potential to protect human keratinocytes against UVB-induced cell damage and death.

Galangin (3,5,7-Trihydroxyflavone) Shields Human Keratinocytes from Ultraviolet B-Induced Oxidative Stress

( Susara Ruwan Kumara Madduma Hewage ),( Mei Jing Piao ),( Ki Cheon Kim ),( Ji Won Cha ),( Xia Han ),( Yung Hyun Choi ),( Sung Wook Chae ),( Jin Won Hyun ) 한국응용약물학회 2015 Biomolecules & Therapeutics(구 응용약물학회지) Vol.23 No.2

Most skin damage caused by ultraviolet B (UVB) radiation is owing to the generation of reactive oxygen species. Phytochemicals can act as antioxidants against UVB-induced oxidative stress. This study investigated the protective effects of the flavone galangin against UVB-induced oxidative damage in human keratinocytes. Galangin efficiently scavenged free radicals and reduced UVBinduced damage to cellular macromolecules, such as DNA, lipids, and proteins. Furthermore, galangin rescued cells undergoing apoptosis induced by UVB radiation via recovering mitochondrial polarization and down-regulating apoptotic proteins. These results showed that galangin protects human keratinocytes against UVB radiation-induced cellular damage and apoptosis via its antioxidant effects.

향나무 추출물의 항균, 항산화 및 각질세포 보호 효과

유민정,이숙영,조상열,박열 한국피부과학연구원 2010 대한피부미용학회지 Vol.8 No.2

This Study was carried out to investigate antimicrobial activity and antioxidative effects of Juniperus chinensis and protective effects on Human HaCaT Keratinocyte. The antimicrobial activity of Juniperus chinensis extracts using a paper disc method and using four human scalp, skin pathogens. The antioxidative of extracts was used the method of DPPH radical, total polyphenolics and flavonoid contents. Protective effects of Juniperus chinensis on HaCaT keratinocytes against oxidative stree induced by hydrogen peroxide were also measured. In our results antimicrobial activity of Juniperus chinensis extracts was the strongest activity on P. acnes. In addition with Juniperus chinensis did show cytotoxicity on HaCat keratinocyte beneath the concentration elevated in dose -dependen. DPPH free radical scavening activities of Juniperus chinensis were elevated in dose dependent manner. In addition, The value of half maximal inhibitiory concentration(IC50) was 153.8±0.49 ㎍/㎖. 84.9 ± 0.01 ㎍/㎖ of total phenolics and 66.4± 0.02 ㎍/㎖ of flavonoids. Finally Juniperus chinensis showed protective effect against cell dath of HaCaT cell tnduced by hydrogen peroxide significantiy. In conclusion. these results suggest that Juniperus chinensis extracts may have antibioticl, antioxantic action on human scalp, skin and sugest the possibility as cosmetic material 향나무 잎 추출물의 두피질환 원인균에 대한 항균효과와 항산화 작용에 중요한 부분을 차지하고 있는 DPPH 자유라디칼 제거능을 측정하였으며, 총 페놀과 플라보노이드 함량을 조사하였다. 또한 인간 유래 피부각질세포인 HaCaT 세포에서 과산화수소에 의하여 발생하는 산화적 스트레스를 효율적으로 제거할 수 있는지 관찰하였다. 실험결과 propionibacterium acnes 에서 항균효과가 가장 우수하였고 DPPH 자유라디칼 제거능은 농도 의존적으로 증가되었다. IC50 값을 구한 결과 153.8±0.49 ㎍/㎖으로 나타났다. 폴리페놀함량을 측정한 결과 84.9 ± 0.01 ㎍/㎖의 함량이 나타냈으며, 플라보노이드는 함량은 66.4± 0.02 ㎍/㎖으로 나타내었다. 과산화수소에 의해 발생하는 산화적 스트레스에 대한 세포사멸을 방지하였다. 이상의 결과 향나무 잎 추출물은 두피, 피부과 영역 사용 확대의 근거로 사용될 수 있다.

UVB protective effect of hydrolysate derived Hermetia illucens in human keratinocytes

안긴내,박서영,Yoo Si-won,이서희,Choi Jong Min,Song Jin Woo,Han Eui Jeong 한국키틴키토산학회 2022 한국키틴키토산학회지 Vol.27 No.2

In this study, we prepared enzymatic extracts using trypsin and pepsin from Hermetia illucens (THI and PHI) and evaluated their effects on oxidative stress in ultraviolet B (UVB)-stimulated keratinocytes. PHI showed a higher extraction yield than THI, but the protein content of THI was higher. When the antioxidant activity was assessed in vitro, both indicated an effect of 25% or more compared to the positive control (L-ascorbic acid). In addition, both THI and PHI decreased the generation of intracellular reactive oxygen species (ROS) in UVB-stimulated keratinocytes, without cytotoxicity at all concentrations used. We explored the efficacy of THI. Results demonstrated that THI effectively reduced apoptosis and sub-G1 DNA content. In addition, it was found that UVB-stimulated keratinocytes protect skin keratinocytes from UVB by regulating the apoptotic pathway, MAPKs, and NF-kB signaling pathways. In conclusion, these results show that trypsin hydrolysate of H. illucens protects cells from oxidative stress in UVB-irradiated human keratinocytes.

Effect of LED Irradiation on Proliferation of Human Epidermal Keratinocyte for Convergence

박정숙,김미혜,이재혁 한국디지털정책학회 2016 디지털융복합연구 Vol.14 No.11

The purpose of this study is to determine the effect of the light-emitting-diode (LED) to investigate proliferation of human epidermal keratinocyte and collagen, procollagen expression. In order to determine whether LED irradiation can safely be applied to human skin, the proliferative effects of LED irradiation were determined by MTS assay in Human Epidermal Keratinocytes. Wavelength of 470nm LED irradiation increased mRNA expression of collagen, procollagen without cytotoxity. Our results suggest that 470nm LED irradiation may have a proliferative effects and collagen synthesis property. In order to determine whether LED irradiation can safely be applied to human skin, the cytotoxic effects of LED irradiation were determined by MTS assay in Human Dermal Fibroblasts (HDF). As far as we know, this is the first report demonstrating in vitro collagen synthesis activity of 470nm LED irradiation and being a scientific basis for the cosmetic.

N-nitroso-N-methylurea and N-nitroso-N-ethylurea Decrease in Nitric Oxide Production in Human Malignant Keratinocytes

Ki-Young Moon 대한의생명과학회 2018 Biomedical Science Letters Vol.24 No.1

N-nitroso-N-methylurea (NMU) and N-nitroso-N-ethylurea (NEU), direct alkylating chemical mutagens and carcinogens, are shown to be the upregulators of cellular NF-κB, regulating various genes that mediate tumorigenesis and carcinogenesis. Nitric oxide (NO), a toxic reactive radical gas, has been known to induce programmed cell death or apoptosis in various cells. Therefore, the assessment of NO production was examined to elucidate the possible contribution of NO release to the chemical carcinogenic potency of NMU and NEU in human skin cells. NMU and NEU did not alter the NO production, but they caused a significant downregulation of the NO generation on lipopolysaccharide (LPS)-induced NO production at concentrations ranging from 2~5 μM. The degree of downregulation of NO by NMU and NEU decreased up to 15% and 20%, respectively, compared to the control. These results demonstrate that the LPS-inducible keratinocytes NO synthase is involved in modulating carcinogenic potency by NMU and NEU, and the regulation of the cellular NF-κB activity by NMU and NEU is negatively correlated with the level of LPS-induced NO production in human skin cells. The findings of this study suggest the hypothesis that NMU and NEU-induced carcinogenesis may be associated with the downregulation of NO production, and the inducible NO may play an important role in NMU and NEU-induced carcinogenicity in human epidermal keratinocytes.