检索结果-内蒙古大学图书馆

Targeting protein-protein interaction interfaces with antiviral N protein inhibitor in SARS-CoV-2

Biophysical Journal 2024年第4期123卷 478-488页

作者： Hong, Jhen-Yi Lin, Shih-Chao Kehn-Hall, Kylene Zhang, Kai-Min Luo, Shun-Yuan Wu, Hung-Yi Chang, Sui-Yuan Hou, Ming-Hon Institute of Genomics and Bioinformatics and Department of Life Sciences National Chung Hsing University Taichung Taiwan Bachelor Degree Program in Marine Biotechnology College of Life Sciences National Taiwan Ocean University Keelung Taiwan Department of Biomedical Sciences and Pathobiology Virginia-Maryland College of Veterinary Medicine Virginia Polytechnic Institute and State University Blacksburg VA United States Center for Emerging Zoonotic and Arthropod-borne Pathogens Virginia Polytechnic Institute and State University Blacksburg VA United States Department of Chemistry National Chung Hsing University Taichung Taiwan Graduate Institute of Veterinary Pathobiology College of Veterinary Medicine National Chung Hsing University. Taichung Taiwan Department of Clinical Laboratory Sciences and Medical Biotechnology College of Medicine National Taiwan University Taipei Taiwan PhD Program in Medical Biotechnology National Chung Hsing University Taichung Taiwan Biotechnology Center National Chung Hsing University Taichung Taiwan

Coronaviruses not only pose significant global public health threats but also cause extensive damage to livestock-based industries. Previous studies have shown that 5-benzyloxygramine (P3) targets the Middle East respiratory syndrome coronavirus (MERS-CoV) nucleocapsid (N) protein N-terminal domain (N-NTD), inducing non-native protein-protein interactions (PPIs) that impair N protein function. Moreover, P3 exhibits broad-spectrum antiviral activity against CoVs. The sequence similarity of N proteins is relatively low among CoVs, further exhibiting notable variations in the hydrophobic residue responsible for non-native PPIs in the N-NTD. Therefore, to ascertain the mechanism by which P3 demonstrates broad-spectrum anti-CoV activity, we determined the crystal structure of the SARS-CoV-2 N-NTD:P3 complex. We found that P3 was positioned in the dimeric N-NTD via hydrophobic contacts. Compared with the interfaces in MERS-CoV N-NTD, P3 had a reversed orientation in SARS-CoV-2 N-NTD. The Phe residue in the MERS-CoV N-NTD:P3 complex stabilized both P3 moieties. However, in the SARS-CoV-2 N-NTD:P3 complex, the Ile residue formed only one interaction with the P3 benzene ring. Moreover, the pocket in the SARS-CoV-2 N-NTD:P3 complex was more hydrophobic, favoring the insertion of the P3 benzene ring into the complex. Nevertheless, hydrophobic interactions remained the primary stabilizing force in both complexes. These findings suggested that despite the differences in the sequence, P3 can accommodate a hydrophobic pocket in N-NTD to mediate a non-native PPI, enabling its effectiveness against various CoVs. © 2024 Biophysical Society

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Rapid Escherichia coli Cloned DNA Detection in Serum Using an Electrical Double Layer-Gated Field-Effect Transistor-Based DNA Sensor

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Analytical Chemistry 2023年第17期95卷 6871-6878页

作者： Paulose, Akhil K. Hou, Yueh-Ju Huang, Yu-Shan Chakkalaparambil Dileep, Navyamol Chiu, Chia-Lin Pal, Arnab Kalaimani, Vishal Mani Lin, Zong-Hong Chang, Chuang-Rung Chen, Cheng-Pin Lin, Yi-Chun Cheng, Chien-Yu Cheng, Shu-Hsing Cheng, Chao-Min Wang, Yu-Lin Institute of Nanoengineering and Microsystems National Tsing Hua University Hsinchu30013 Taiwan Department of Power Mechanical Engineering National Tsing Hua University Hsinchu30013 Taiwan College of Semiconductor Research National Tsing Hua University Hsinchu30013 Taiwan Department of Life Sciences National University of Kaohsiung Kaohsiung811726 Taiwan Department of Biomedical Engineering National Taiwan University Taipei10617 Taiwan Institute of Biotechnology National Tsing Hua University Hsinchu30013 Taiwan Department of Infectious Diseases Taoyuan General Hospital Ministry of Health and Welfare Taoyuan32748 Taiwan Institute of Biomedical Engineering National Tsing Hua University Hsinchu30013 Taiwan International Intercollegiate PhD Program National Tsing Hua University Hsinchu30013 Taiwan

In this study, a rapid diagnosis platform was developed for the detection of Escherichia coli O157:H7. An electrical double layer (EDL)-gated field-effect transistor-based biosensor (BioFET) as a point-of-care testing device is demonstrated with its high sensitivity, portability, high selectivity, quick response, and ease of use. The specially designed ssDNA probe was immobilized on the extended gate electrode to bind the target complementary DNA segment of E. coli, resulting in a sharp drain current change within minutes. The limit of detection for target DNA is validated to a concentration of 1 fM in buffer solution and serum. Meanwhile, the results of a Kelvin probe force microscope were shown to have reduced surface potential of the DNA immobilized sensors before and after the cDNA detection, which is consistent with the decreased drain current of the BioFET. A 1.2 kb E. coli duplex DNA synthesized in plasmid was sonicated and detected in serum samples with the sensor array. Gel electrophoresis was used to confirm the efficiency of sonication by elucidating the length of DNA. Those results show that the EDL-gated BioFET system is a promising platform for rapid identification of pathogens for future clinical needs. © 2023 American Chemical Society

关键词： DNA

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Ellagic acid protects against angiotensin II-induced hypertrophic responses through ROS-mediated MAPK pathway in H9c2 cells

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Environmental Toxicology 2024年第5期39卷 3253-3263页

作者： Lee, Ya-Che Jou, Yeong-Chin Chou, Wan-Ching Tsai, Kun-Ling Shen, Cheng-Huang Lee, Shin-Da Department of Urology Ditmanson Medical Foundation Chiayi Christian Hospital Chia-Yi City Taiwan Department of Urology St. Martin De Porres Hospital Chia-Yi City Taiwan Department of Health and Nutrition Biotechnology College of Medical and Health Science Asia University Taichung City Taiwan Department of Physical Therapy College of Medicine National Cheng Kung University Tainan City Taiwan Institute of Allied Health Science College of Medicine National Cheng Kung University Tainan City Taiwan Department of Biomedical Sciences National Chung Cheng University Chia-Yi Min Hsiung Taiwan Department of Physical Therapy College of Medical and Health Science Asia University Taichung City Taiwan Department of Physical Therapy PhD program in Healthcare Science China Medical University Taichung Taiwan

The early myocardial response of hypertension is an elevation of angiotensin-II (Ang-II) concentration, leading to heart failure and cardiac hypertrophy. This hypertrophic event of the heart is mediated by the interaction of Ang type 1 receptors (AT-R1), thereby modulating NADPH oxidase activity in cardiomyocytes, which alters redox status in cardiomyocytes. Ellagic acid (EA) has anti-inflammatory and anti-oxidative capacities. Thus, EA has potential preventive effects on cardiovascular diseases and diabetes. In the last decades, because the protective effect of EA on Ang-II-induced hypertrophic responses is unclear, this study aims to investigate the protective effect of EA in cardiomyocytes. H9c2 cells were treated to Ang-II 1 μM for 24 h to induce cellular damage. We found that EA protected against Ang-II-increased cell surface area and pro-hypertrophic gene expression in H9c2. EA reduced Ang-II-caused AT-R1 upregulation, thereby inhibiting oxidative stress NADPH oxidase activation. EA mitigated Ang-II-enhanced p38 and extracellular-signal-regulated kinase (ERK) phosphorylation. Moreover, EA treatment under Ang-II stimulation also reversed NF-κB activity and iNOS expression. This study shows that EA protects against Ang-II-induced myocardial hypertrophy and attenuates oxidative stress through reactive oxygen species-mediated mitogen-activated protein kinase signaling pathways in H9c2 cells. Thus, EA may be an effective compound for preventing Ang-II-induced myocardial hypertrophy. © 2024 Wiley Periodicals LLC.

关键词： Oxidative stress

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Hericium erinaceus extracts promote neuronal differentiation and excitability through nootropic metabolite activity

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Biomedicine and Pharmacotherapy 2025年 188卷 118204-118204页

作者： Brandalise, Federico Priori, Erica Cecilia Giammello, Francesca Venuti, MariaTeresa Ratto, Daniela Goppa, Lorenzo Locatelli, Carlo Alessandro Savino, Elena Roda, Elisa Rossi, Paola Department of Biomedical Sciences Division of Neuroscience and Clinical Pharmacology University of Cagliari Cittadella Universitaria di Monserrato Monserrato 09042 Italy Department of Biology and Biotechnology “L. Spallanzani” University of Pavia Pavia 27100 Italy PhD Program in Genetics Molecular and Cellular Biology University of Pavia Italy Department of Earth and Environmental Sciences (DSTA) University of Pavia Pavia 27100 Italy PhD Program in Earth and Environmental Sciences University of Pavia Italy Laboratory of Clinical & Experimental Toxicology Pavia Poison Centre National Toxicology Information Centre Toxicology Unit Istituti Clinici Scientifici Maugeri IRCCS Pavia Pavia 27100 Italy

This study investigates the effects of a blend of Hericium erinaceus (lion's mane mushroom) extract on the differentiation of SH-SY5Y cells, a human neuroblastoma cell line, revealing potential therapeutic implications for neuroblastoma management. Treatment with this blend induced cells differentiation towards a neuron-like profile, as evidenced by enhanced neuronal excitability and upregulation of neuronal markers, such as βIII-tubulin and synaptotagmin. Additionally, the treatment significantly reduced PCNA, a key regulator of proliferation, alongside a decrease in stemness markers, indicating a shift toward a more mature and less proliferative phenotype. These findings demonstrate the ability of Hericium erinaceus to promote neuronal differentiation and inhibit proliferation in neuroblastoma cells, highlighting its therapeutic potential for managing neuroblastoma and potentially other neurological disorders. The results suggest that Hericium erinaceus may serve as a promising candidate for the development of novel neuroregenerative therapies. © 2025

关键词： Action potential Cell proliferation Erinacines Hericenones Hericium erinaceus Neuroblastoma Neuronal differentiation Patch clamp SH-SY5Y Voltage-Gated Sodium Channel Voltage-gated sodium currents

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Enhanced IGF-IIRα Expression Exacerbates Lipopolysaccharide-Induced Cardiac Inflammation, Hypertrophy, and Apoptosis Through Calcineurin Activation

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Environmental Toxicology 2024年第11期39卷 5173-5186页

作者： Boonha, Khwanchit Kuo, Wei-Wen Tsai, Bruce Chi-Kang Hsieh, Dennis Jine-Yuan Lin, Kuan-Ho Lu, Shang-Yeh Kuo, Chia-Hua Yang, Liang-Yo Huang, Chih-Yang Graduate Institute of Biomedical Sciences China Medical University Taichung Taiwan Cardiovascular and Mitochondrial Related Disease Research Center Hualien Tzu Chi Hospital Buddhist Tzu Chi Medical Foundation Hualien Taiwan Faculty of Tropical Medicine Mahidol University Bangkok Thailand Department of Biological Science and Technology College of Life Sciences China Medical University Taichung Taiwan PhD Program for Biotechnology Industry China Medical University Taichung Taiwan School of Pharmacy China Medical University Taichung Taiwan Department of Medical Laboratory and Biotechnology Chung Shan Medical University Taichung Taiwan Clinical Laboratory Chung Shan Medical University Hospital Taichung Taiwan Department of Emergency Medicine China Medical University Hospital Taichung Taiwan College of Medicine China Medical University Taichung Taiwan Division of Cardiovascular Medicine Department of Internal Medicine China Medical University Hospital Taichung Taiwan Laboratory of Exercise Biochemistry University of Taipei Taipei Taiwan Institute of Sports Sciences University of Taipei Taipei Taiwan Department of Kinesiology and Health Science College of William and Mary WilliamsburgVA United States School of Physical Education and Sports Science Soochow University Suzhou China Department of Physiology School of Medicine College of Medicine China Medical University Taichung Taiwan Laboratory for Neural Repair China Medical University Hospital Taichung Taiwan Department of Medical Research China Medical University Hospital China Medical University Taichung Taiwan Department of Medical Laboratory Science and Biotechnology Asia University Taichung Taiwan Center of General Education Buddhist Tzu Chi Medical Foundation Tzu Chi University of Science and Technology Hualien Taiwan

Cardiovascular disease is one of the leading causes of death worldwide and has a high prevalence. Insulin-like growth factor-II receptor α (IGF-IIRα) acts as a stress-inducible negative regulator. This study focused on the substantial impact of heightened expression of IGF-IIRα in cardiac myoblasts and its association with the exacerbation of cardiac dysfunction. Using lipopolysaccharide (LPS)-induced H9c2 cardiac myoblasts as a model for sepsis, we aimed to elucidate the molecular interactions between IGF-IIRα and LPS in exacerbating cardiac injury. Our findings demonstrated a synergistic induction of cardiac inflammation and hypertrophy by LPS stimulation and IGF-IIRα overexpression, leading to decreased cell survival. Excessive calcineurin activity, triggered by this combined condition, was identified as a key factor exacerbating the negative effects on cell survival. Cellular changes such as cell enlargement, disrupted actin filaments, and upregulation of hypertrophy-related and inflammation-related proteins contributed to the overall hypertrophic and inflammatory responses. Overexpression of IGF-IIRα also exacerbated apoptosis induced by LPS in H9c2 cardiac myoblasts. Inhibiting calcineurin in LPS-treated H9c2 cardiac myoblasts with IGF-IIRα overexpression effectively reversed the detrimental effects, reducing cell damage and mitigating apoptosis-related cardiac mechanisms. Our study suggests that under sepsis-like conditions in the heart with IGF-IIRα overexpression, hyperactivation of calcineurin worsens cardiac damage. Suppressing IGF-IIRα and calcineurin expression could be a potential intervention to alleviate the impact of the illness and improve cardiac function. © 2024 Wiley Periodicals LLC.

关键词： Heart

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Ubiquitin carboxyl-terminal hydrolase 11 promotes autophagy by de-ubiquitinating and stabilizing Beclin-1

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Genome Instability & Disease 2022年第1期3卷 47-55页

作者： Li, Zheng Rao, Shaohong Song, Chunwei Zhu, Min Zhao, Hongchang Yuan, Shuping Peng, Bin Xu, Xingzhi Guangdong Key Laboratory for Genome Stability & Disease Prevention and Carson International Cancer Center Marshall Laboratory of Biomedical Engineering Shenzhen University School of Medicine Shenzhen China College of Life Sciences Capital Normal University Beijing China Institute of Translational Medicine College of Medicine Qingdao University Qingdao China Shenzhen University-Friedrich Schiller Universität Jena Joint PhD Program in Biomedical Sciences Shenzhen University School of Medicine Shenzhen China

Autophagy is a major degradation process that degrades and recycles cytoplasmic materials through lysosome for maintaining cellular homeostasis. Dysregulated autophagy is linked with numerous human diseases including cancer. Autophagy marker protein B-cell lymphoma-2 interacting protein 1 (Beclin-1) is essential for autophagosome initiation and maturation. Recently, Ubiquitin carboxyl-terminal hydrolase 11 (USP11) has been reported to promote or inhibit autophagy without identification of any direct target. Here through biochemical reaction in vitro, we demonstrate that USP11 directly interacts with Beclin-1. Both in vitro and in vivo de-ubiquitination assays revealed that USP11 de-ubiquitinates Beclin-1. USP11-mediated de-ubiquitination stabilized Beclin-1 and enhanced the formation of the autophagy-specific class III phosphatidylinositol 3-kinase complexes 1 and 2, thereby promoting autophagy. Together, our results demonstrated that USP11 promotes autophagy under unperturbed conditions by de-ubiquitinating and stabilizing Beclin-1 which may serve as a therapeutic target for autophagy-related diseases.

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Demethoxymurrapanine, an indole-naphthoquinone alkaloid, inhibits the proliferation of oral cancer cells without major side effects on normal cells

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Environmental Toxicology 2024年第3期39卷 1221-1234页

作者： Chuang, Ya-Ting Yen, Ching-Yu Shiau, Jun-Ping Chang, Fang-Rong Duh, Chang-Yih Sung, Ping-Jyun Chen, Kuan-Liang Tsai, Yi-Hong Tang, Jen-Yang Jeng, Jiiang-Huei Sheu, Jyh-Horng Chang, Hsueh-Wei Department of Biomedical Science and Environmental Biology PhD Program in Life Sciences College of Life Science Kaohsiung Medical University Kaohsiung Taiwan School of Dentistry Taipei Medical University Taipei Taiwan Department of Oral and Maxillofacial Surgery Chi-Mei Medical Center Tainan Taiwan Division of Breast Oncology and Surgery Department of Surgery Kaohsiung Medical University Hospital Kaohsiung Medical University Kaohsiung Taiwan Graduate Institute of Natural Products Kaohsiung Medical University Kaohsiung Taiwan Department of Marine Biotechnology and Resources National Sun Yat-sen University Kaohsiung Taiwan National Museum of Marine Biology and Aquarium Pingtung Taiwan Department of Pharmacy and Master Program College of Pharmacy and Health Care Tajen University Pingtung Taiwan School of Post-Baccalaureate Medicine Kaohsiung Medical University Kaohsiung Taiwan Department of Radiation Oncology Kaohsiung Medical University Hospital Kaohsiung Medical University Kaohsiung Taiwan School of Dentistry College of Dental Medicine Kaohsiung Medical University Kaohsiung Taiwan Department of Dentistry Kaohsiung Medical University Hospital Kaohsiung Taiwan Department of Dentistry National Taiwan University Hospital Taipei Taiwan Department of Medical Research China Medical University Hospital China Medical University Taichung Taiwan Department of Medical Research Kaohsiung Medical University Hospital Kaohsiung Taiwan Center for Cancer Research Kaohsiung Medical University Kaohsiung Taiwan

Antioral cancer drugs need a greater antiproliferative impact on cancer than on normal cells. Demethoxymurrapanine (DEMU) inhibits proliferation in several cancer cells, but an in-depth investigation was necessary. This study evaluated the proliferation-modulating effects of DEMU, focusing on oral cancer and normal cells. DEMU (0, 2, 3, and 4 μg/mL) at 48 h treatments inhibited the proliferation of oral cancer cells (the cell viability (%) for Ca9-22 cells was 100.0 ± 2.2, 75.4 ± 5.6, 26.0 ± 3.8, and 15.4 ± 1.4, and for CAL 27 cells was 100.0 ± 9.4, 77.2 ± 5.9, 57.4 ± 10.7, and 27.1 ± 1.1) more strongly than that of normal cells (the cell viability (%) for S-G cells was 100.0 ± 6.6, 91.0 ± 4.6, 95.0 ± 2.6, and 95.8 ± 5.5), although this was blocked by the antioxidant N-acetylcysteine. The presence of oxidative stress was evidenced by the increase of reactive oxygen species and mitochondrial superoxide and the downregulation of the cellular antioxidant glutathione in oral cancer cells, but these changes were minor in normal cells. DEMU also caused greater induction of the subG1 phase, extrinsic and intrinsic apoptosis (annexin V and caspases 3, 8, and 9), and DNA damage (γH2AX and 8-hydroxy-2-deoxyguanosine) in oral cancer than in normal cells. N-acetylcysteine attenuated all these DEMU-induced changes. Together, these data demonstrate the preferential antiproliferative function of DEMU in oral cancer cells, with the preferential induction of oxidative stress, apoptosis, and DNA damage in these cancer cells, and low cytotoxicity toward normal cells. © 2023 Wiley Periodicals LLC.

关键词： Oxidative stress

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Development of a respiratory sound labeling software for training a deep learning-based respiratory sound analysis model

Development of a respiratory sound labeling software for tra...

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International Forum on Medical Imaging in Asia 2021, IFMIA 2021

作者： Hsu, Fu-Shun Huang, Chao-Jung Kuo, Chen-Yi Huang, Shang-Ran Cheng, Yuan-Ren Wang, Jia-Horng Wu, Yi-Lin Tzeng, Tzu-Ling Lai, Feipei Graduate Institute of Biomedical Electronics and Bioinformatics National Taiwan University Taipei Taiwan Heroic Faith Medical Science Corporation Limited New Taipei Taiwan National Taiwan University-Stanford Joint Program Office of AI in Biotechnology Ministry of Science and Technology Joint Research Center for Artificial Intelligence Technology and All Vista Healthcare Taipei Taiwan Department of Life Science College of Life Science National Taiwan University Taipei Taiwan Institute of Biomedical Sciences Academia Sinica Taipei Taiwan Department of Critical Care Far Eastern Memorial Hospital New Taipei City Taiwan

ISBN: (数字)9781510644212

ISBN: (纸本)9781510644205

Respiratory auscultation can help healthcare professionals detect abnormal respiratory conditions if adventitious lung sounds are heard. The state-of-the-art artificial intelligence technologies based on deep learning show great potential in the development of automated respiratory sound analysis. To train a deep learning-based model, a huge number of accurate labels of normal breath sounds and adventitious sounds are needed. In this paper, we demonstrate the work of developing a respiratory sound labeling software to help annotators identify and label the inhalation, exhalation, and adventitious respiratory sound more accurately and quickly. Our labeling software integrates six features from MATLAB Audio Labeler, and one commercial audio editor, RX7. As of October, 2019, we have labeled 9,765 15- second-long audio files of breathing lung sounds, and accrued 34,095 inhalation labels,18,349 exhalation labels, 13,883 continuous adventitious sounds (CASs) labels and 15,606 discontinuous adventitious sounds (DASs) labels, which are significantly larger than previously published studies. The trained convolutional recurrent neural networks based on these labels showed good performance with F1-scores of 86.0% on inhalation event detection, 51.6% on CASs event detection and 71.4% on DASs event detection. In conclusion, our results show that our proposed respiratory sound labeling software could easily pre-define a label, perform one-click labeling, and overall facilitate the process of accurately labeling. This software helps develop deep learning-based models that require a huge amount of labeled acoustic data. © 2021 SPIE.

关键词： Recurrent neural networks

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Capnesterones A and B, new steroids isolated from the soft coral Capnella imbricata

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Phytochemistry Letters 2024年 61卷 115-119页

作者： Cheng, Ting-Yu Yang, Chia-Jung Chen, Po-Jen Chen, Yu-Li Peng, Bo-Rong Kung, Te-An Wen, Zhi-Hong Lai, Kuei-Hung Chung, Hsu-Ming Department of Applied Chemistry National Pingtung University Pingtung 900393 Taiwan Clinical Immunology Laboratory Department of Medical Research E-Da Hospital I-Shou University Kaohsiung 824005 Taiwan Research Center for Chinese Herbal Medicine Graduate Institute of Healthy Industry Technology College of Human Ecology Chang Gung University of Science and Technology Taoyuan 33305 Taiwan Graduate Institute of Biomedical Sciences Graduate Institute of Natural Products College of Medicine Chang Gung University Taoyuan 33305 Taiwan Graduate Institute of Pharmacognosy College of Pharmacy Taipei Medical University Taipei 11031 Taiwan Institute of Food Safety Management National Pingtung University of Science and Technology Pingtung 91201 Taiwan Department of Marine Biotechnology and Resources National Sun Yat-sen University Kaohsiung 804201 Taiwan Institute of BioPharmaceutical Sciences National Sun Yat-sen University Kaohsiung 804201 Taiwan PhD Program in Clinical Drug Development of Herbal Medicine College of Pharmacy Taipei Medical University Taipei 11031 Taiwan Traditional Herbal Medicine Research Center Taipei Medical University Hospital Taipei 11031 Taiwan

Two new non-withanolidal steroids with a cross-conjugated dienone structural unit in ring A, capnesterones A (1) and B (2), were isolated from the soft coral Capnella imbricata in the southeast waters of Taiwan. The compounds' structures were elucidated through a comprehensive analysis of spectroscopic data, including IR, ESIMS, ¹H NMR, and ¹³C NMR, and through comparison with previous literature. At a concentration of 10 μM, steroids 1 and 2 exhibited reductions in iNOS levels (60 % and 82 %) compared to LPS-stimulated control cells, along with increased release (134 % and 110 %) against COX-2 protein expressions. © 2024 Phytochemical Society of Europe

关键词： Capnella imbricata COX-2 INOS Steroid

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Low-dose imiquimod induces melanogenesis in melanoma cells through an ROS-mediated pathway

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Journal of Dermatological Science 2024年第1期113卷 18-25页

作者： Li, Zheng-Yi Chang, Shu-Hao Liu, Kuang-Ting Wu, Alaina Edelie Hsu, Chien-Sheng Huang, Shi-Wei Chung, Mu-Chi Wang, Shih-Chung Kao, Jun-Kai Chen, Yi-Ju Shieh, Jeng-Jer Institute of Biomedical Sciences National Chung Hsing University Taichung Taiwan Department of Pathology & Laboratory Medicine Taoyuan Armed Forces General Hospital Taoyuan Taiwan Frontier Molecular Medical Research Center in Children Changhua Christian Children Hospital Changhua County Taiwan Center for Cell Therapy and Translation Research China Medical University Hospital Taichung Taiwan Division of Nephrology Department of Medicine Taichung Veterans General Hospital Taichung Taiwan PhD Program in Translational Medicine National Chung Hsing University Taichung Taiwan Department of Biotechnology Asia University Taichung Taiwan Rong Hsing Research Center for Translational Medicine National Chung Hsing University Taichung Taiwan Division of Pediatric Hematology/Oncology Changhua Christian Children Hospital Changhua County Taiwan Department of Post-Baccalaureate Medicine College of Medicine National Chung Hsing University Taichung Taiwan Department of Dermatology Taichung Veterans General Hospital Taichung Taiwan Department of Education and Research Taichung Veterans General Hospital Taichung Taiwan

Background: Melanogenesis is the process of melanin maturation which not only protects skin from UV radiation but also plays an important role in antigenicity of melanomas. Imiquimod (IMQ) is a toll-like receptor 7 (TLR7) agonist that exhibits antiviral and anticancer activity. Objective: To explore whether IMQ could induce melanogenesis in melanoma cells. Methods: The mouse melanoma cell line B16F10, the mouse immortalized melanocyte Melan-A, and human melanoma cell lines MNT-1, C32 and A375 were utilized in this study. The pigmented level was observed by the centrifuged cell pellet. The intracellular and extracellular melanin levels were examined in the absorbance in NaOH-extracted cell lysate and cell-cultured medium, respectively. The expression of melanogenesis related proteins was examined by immunoblotting. The intracellular cyclic AMP amount was evaluated by the cAMP Glo assay kit. The activity of phosphodiesterase 4B (PDE4B) was investigated by CREB reporter assay with overexpressed PDE4B or not. Results: We demonstrated that a low dose of IMQ could trigger melanogenesis in B16F10 cells. IMQ induced microphthalmia-associated transcription factor (MITF) nuclear translocation, upregulated the expression of melanogenesis-related proteins, increased tyrosinase (TYR) activity, and led to pigmentation in B16F10 cells. Next, we found that IMQ-induced melanogenesis was activated by excessive intracellular cAMP accumulation, which was regulated through IMQ-mediated PDE4B inhibition. Finally, IMQ-induced ROS production was found to be involved in melanogenesis by its control of PDE4B activity. Conclusions: Low dose of IMQ could activate melanogenesis through the ROS/PDE4B/PKA pathway in melanoma cells. © 2023 Japanese Society for Investigative Dermatology

关键词： Imiquimod Melanogenesis Reactive oxygen species

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