Kuei-Hung Lai1, Po-Jen Chen2, Chih-Chuan Chen3, Sien-Hung Yang4, Mohamed El-Shazly5, Yu-Chia Chang6, Yi-Hsuan Wu7, Yi-Hsiu Wu8, Yi-Hsuan Wang9, Hsi-Lung Hsieh10, Tsong-Long Hwang11. 1. Research Center for Chinese Herbal Medicine, Graduate Institute of Health Industry Technology, College of Human Ecology, Chang Gung University of Science and Technology, Taoyuan, 33303, Taiwan; Graduate Institute of Pharmacognosy, College of Pharmacy, Taipei Medical University, Taipei, 11031, Taiwan. Electronic address: mos19880822@gmail.com. 2. Department of Cosmetic Science, Providence University, Taichung, 43301, Taiwan. Electronic address: litlep@hotmail.com. 3. Graduate Institute of Natural Products, School of Traditional Chinese Medicine, Chang Gung University, Taoyuan, 33302, Taiwan. Electronic address: pariede@gmail.com. 4. Center for Traditional Chinese Medicine, Chang Gung Memorial Hospital, Taoyuan, 33305, Taiwan; School of Traditional Chinese Medicine, Chang Gung University, Taoyuan, 33302, Taiwan. Electronic address: dryang@mail.cgu.edu.tw. 5. Department of Pharmacognosy and Natural Products Chemistry, Faculty of Pharmacy, Ain-Shams University, Organization of African Unity Street, Abassia, Cairo, 11566, Egypt; Department of Pharmaceutical Biology, Faculty of Pharmacy and Biotechnology, German University in Cairo, Cairo, 11432, Egypt. Electronic address: mohamed.elshazly@pharma.asu.edu.eg. 6. Research Center for Chinese Herbal Medicine, Graduate Institute of Health Industry Technology, College of Human Ecology, Chang Gung University of Science and Technology, Taoyuan, 33303, Taiwan. Electronic address: ycchang03@mail.cgust.edu.tw. 7. Research Center for Chinese Herbal Medicine, Graduate Institute of Health Industry Technology, College of Human Ecology, Chang Gung University of Science and Technology, Taoyuan, 33303, Taiwan. Electronic address: yhwu03@mail.cgust.edu.tw. 8. Graduate Institute of Natural Products, School of Traditional Chinese Medicine, Chang Gung University, Taoyuan, 33302, Taiwan. Electronic address: modemtw@gmail.com. 9. Graduate Institute of Natural Products, School of Traditional Chinese Medicine, Chang Gung University, Taoyuan, 33302, Taiwan. Electronic address: e0919133641@gmail.com. 10. Research Center for Chinese Herbal Medicine, Graduate Institute of Health Industry Technology, College of Human Ecology, Chang Gung University of Science and Technology, Taoyuan, 33303, Taiwan; Department of Nursing, Division of Basic Medical Sciences, Chang Gung University of Science and Technology, Taoyuan, 33303, Taiwan; Department of Neurology, Chang Gung Memorial Hospital, Taoyuan, 33305, Taiwan. Electronic address: hlhsieh@mail.cgust.edu.tw. 11. Research Center for Chinese Herbal Medicine, Graduate Institute of Health Industry Technology, College of Human Ecology, Chang Gung University of Science and Technology, Taoyuan, 33303, Taiwan; Graduate Institute of Natural Products, School of Traditional Chinese Medicine, Chang Gung University, Taoyuan, 33302, Taiwan; Research Center for Food and Cosmetic Safety, College of Human Ecology, Chang Gung University of Science and Technology, Taoyuan, 33303, Taiwan; Department of Anaesthesiology, Chang Gung Memorial Hospital, Taoyuan, 33305, Taiwan; Department of Chemical Engineering, Ming Chi University of Technology, New Taipei City, 24301, Taiwan. Electronic address: htl@mail.cgu.edu.tw.
Abstract
ETHNOPHARMACOLOGICAL RELEVANCE: Lophatherum gracile Brongn. (L. gracile) has been long used in traditional herbal medicine to clinically clear heat, disinhibit dampness, and treat inflammation. However, the effect of L. gracile on the activation of human neutrophils remains unclear. AIM OF THE STUDY: The aim of current study is to investigate the anti-inflammatory properties of L. gracile extract (LGE) in N-formyl-methionyl-leucyl-phenylalanine (fMLF)-induced activation of human neutrophils. MATERIALS AND METHODS: Superoxide anion generation and elastase release were estimated by spectrophotometry. A series of signaling pathways including mitogen-activated protein kinases (MAPKs) and protein kinase B (Akt), as well as calcium mobilization were studied by Western blot analysis and spectrofluorometry. RESULTS: Our experimental results indicated that the nontoxic dosage of LGE does-dependently inhibited the fMLF-induced superoxide anion (O2•-) generation, elastase release, CD11b expression, adhesion, and chemotactic migration in human neutrophils. LGE selectively inhibited the fMLF-induced phosphorylation of JNK but not p38, ERK, or Akt in human neutrophils. LGE also decreased the intracellular Ca2+ levels ([Ca2+]i) in fMLF-activated human neutrophils. However, a specific JNK inhibitor inhibited the fMLF-induced O2•- generation and CD11b expression, but it had no effect on [Ca2+]i in human neutrophils. CONCLUSIONS: LGE exhibited anti-inflammatory activities in fMLF-activated human neutrophils. The pharmacological mechanisms of LGE-repressed neutrophilic inflammation were through two independent pathways, JNK signaling and calcium mobilization. Our results suggested that LGE holds the potential to be developed as an anti-inflammatory botanical medicine.
ETHNOPHARMACOLOGICAL RELEVANCE: Lophatherum gracile Brongn. (L. gracile) has been long used in traditional herbal medicine to clinically clear heat, disinhibit dampness, and treat inflammation. However, the effect of L. gracile on the activation of human neutrophils remains unclear. AIM OF THE STUDY: The aim of current study is to investigate the anti-inflammatory properties of L. gracile extract (LGE) in N-formyl-methionyl-leucyl-phenylalanine (fMLF)-induced activation of human neutrophils. MATERIALS AND METHODS:Superoxide anion generation and elastase release were estimated by spectrophotometry. A series of signaling pathways including mitogen-activated protein kinases (MAPKs) and protein kinase B (Akt), as well as calcium mobilization were studied by Western blot analysis and spectrofluorometry. RESULTS: Our experimental results indicated that the nontoxic dosage of LGE does-dependently inhibited the fMLF-induced superoxide anion (O2•-) generation, elastase release, CD11b expression, adhesion, and chemotactic migration in human neutrophils. LGE selectively inhibited the fMLF-induced phosphorylation of JNK but not p38, ERK, or Akt in human neutrophils. LGE also decreased the intracellular Ca2+ levels ([Ca2+]i) in fMLF-activated human neutrophils. However, a specific JNK inhibitor inhibited the fMLF-induced O2•- generation and CD11b expression, but it had no effect on [Ca2+]i in human neutrophils. CONCLUSIONS: LGE exhibited anti-inflammatory activities in fMLF-activated human neutrophils. The pharmacological mechanisms of LGE-repressed neutrophilic inflammation were through two independent pathways, JNK signaling and calcium mobilization. Our results suggested that LGE holds the potential to be developed as an anti-inflammatory botanical medicine.