Yong-Jin Kwon1, Eun-Bi Seo2, Seul-Ki Kim3, Kum Hee Noh4, Haeri Lee5, Yeo-Won Joung6, Hyun Mu Shin7, Young-Ah Jang8, Yu Mi Kim9, Jin-Tae Lee10, Sang-Kyu Ye11. 1. Department of Pharmacology and Biomedical Sciences, Seoul National University College of Medicine, Seoul, 03080, South Korea; Biomedical Science Project (BK21PLUS), Seoul National University College of Medicine, Seoul, 03080, South Korea. Electronic address: pistolhunter@snu.ac.kr. 2. Department of Pharmacology and Biomedical Sciences, Seoul National University College of Medicine, Seoul, 03080, South Korea; Biomedical Science Project (BK21PLUS), Seoul National University College of Medicine, Seoul, 03080, South Korea. Electronic address: lime872@snu.ac.kr. 3. Department of Pharmacology and Biomedical Sciences, Seoul National University College of Medicine, Seoul, 03080, South Korea; Biomedical Science Project (BK21PLUS), Seoul National University College of Medicine, Seoul, 03080, South Korea. Electronic address: cielryoma@snu.ac.kr. 4. Department of Pharmacology and Biomedical Sciences, Seoul National University College of Medicine, Seoul, 03080, South Korea; Biomedical Science Project (BK21PLUS), Seoul National University College of Medicine, Seoul, 03080, South Korea. Electronic address: nkhys1209@snu.ac.kr. 5. Department of Pharmacology and Biomedical Sciences, Seoul National University College of Medicine, Seoul, 03080, South Korea. Electronic address: hrlee519@snu.ac.kr. 6. Department of Cosmeceutical Science, Daegu Haany University, Gyeongsan, 38578, South Korea. Electronic address: yeowon0218@naver.com. 7. Wide River Institute of Immunology, Seoul National University, Hongcheon, 25159, South Korea. Electronic address: hyunmu.shin@snu.ac.kr. 8. Convergence Research Center for Smart Healthcare of KS R & DB Foundation, Kyungsung University, Busan, 48434, South Korea. Electronic address: yaviol@ks.ac.kr. 9. Binotec Co., Ltd, Daegu, 42149, South Korea. Electronic address: yandme77@gmail.com. 10. Department of Cosmetic Science, Kyungsung University, Busan, 48434, South Korea. Electronic address: kosmetics@hanmail.net. 11. Department of Pharmacology and Biomedical Sciences, Seoul National University College of Medicine, Seoul, 03080, South Korea; Biomedical Science Project (BK21PLUS), Seoul National University College of Medicine, Seoul, 03080, South Korea; Wide River Institute of Immunology, Seoul National University, Hongcheon, 25159, South Korea; Ischemic/Hypoxic Disease Institute, Seoul National University College of Medicine, Seoul, 03080, South Korea; Neuro-Immune Information Storage Network Research Center, Seoul National University College of Medicine, Seoul, 03080, South Korea. Electronic address: sangkyu@snu.ac.kr.
Abstract
ETHNOPHARMACOLOGICAL RELEVANCE: Chamaecyparis obtusa (Siebold & Zucc.) Endl. (C. obtusa) has been used as folk medicine in East Asia and has been reported to alleviate inflammatory diseases. However, the detailed mechanisms for the anti-inflammatory effects of C. obtusa remain unclear. AIM OF THE STUDY: Although the anti-inflammatory mechanisms of natural products have been studied for decades, it is still important to identify the potential anti-inflammatory effects of natural sources. In this study, we investigated the anti-inflammatory effects and underlying mechanism of C. obtusa leaf extracts. MATERIAL & METHODS: The cell viability was determined by MTT and crystal violet staining. NO production in the supernatant was measured using Griess reagent. The cell lysates were analyzed by immunoblotting and RT-qPCR. Secreted cytokines were analyzed using ELISA kit and cytokine array kit. mRNA expression from the GSE9632 database set. Z-scores were calculated for each gene and visualized by heat map. RESULTS: Among the extracts of C. obtusa obtained with different extraction methods, the 99% ethanol leaf extract (CO99EL) strongly inhibited lipopolysaccharide (LPS)-induced inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression and Janus kinase/signaling transducer and activator of transcription (JAK/STAT) phosphorylation in RAW264.7 cells. In addition, CO99EL strongly inhibited LPS-induced interleukin (IL)-1β, IL-6, IL-27, and C-C motif chemokine ligand (CCL)-1 production and directly inhibited LPS-induced JAK/STAT phosphorylation in RAW264.7 cells. CONCLUSIONS: These findings demonstrate that CO99EL significantly prevents LPS-induced macrophage activation by inhibiting the JAK/STAT axis. Therefore, we suggest the use of C. obtusa extracts as therapeutic approach for inflammatory diseases.
ETHNOPHARMACOLOGICAL RELEVANCE: Chamaecyparis obtusa (Siebold & Zucc.) Endl. (C. obtusa) has been used as folk medicine in East Asia and has been reported to alleviate inflammatory diseases. However, the detailed mechanisms for the anti-inflammatory effects of C. obtusa remain unclear. AIM OF THE STUDY: Although the anti-inflammatory mechanisms of natural products have been studied for decades, it is still important to identify the potential anti-inflammatory effects of natural sources. In this study, we investigated the anti-inflammatory effects and underlying mechanism of C. obtusa leaf extracts. MATERIAL & METHODS: The cell viability was determined by MTT and crystal violet staining. NO production in the supernatant was measured using Griess reagent. The cell lysates were analyzed by immunoblotting and RT-qPCR. Secreted cytokines were analyzed using ELISA kit and cytokine array kit. mRNA expression from the GSE9632 database set. Z-scores were calculated for each gene and visualized by heat map. RESULTS: Among the extracts of C. obtusa obtained with different extraction methods, the 99% ethanol leaf extract (CO99EL) strongly inhibited lipopolysaccharide (LPS)-induced inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression and Janus kinase/signaling transducer and activator of transcription (JAK/STAT) phosphorylation in RAW264.7 cells. In addition, CO99EL strongly inhibited LPS-induced interleukin (IL)-1β, IL-6, IL-27, and C-C motif chemokine ligand (CCL)-1 production and directly inhibited LPS-induced JAK/STAT phosphorylation in RAW264.7 cells. CONCLUSIONS: These findings demonstrate that CO99EL significantly prevents LPS-induced macrophage activation by inhibiting the JAK/STAT axis. Therefore, we suggest the use of C. obtusa extracts as therapeutic approach for inflammatory diseases.