Demonstration of endothelin (ET) receptors on cultured rabbit chondrocytes and stimulation of DNA synthesis and calcium influx by ET-1 via its receptors |
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| Authors: | Akihiro Kinoshita Tomoo Tamura Chiharu Aoki Tohru Nakanishi Shizuo Sobue Fujio Suzuki Kojiro Takahashi Masaharu Takigawa |
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| Affiliation: | 1. Department of Orthopaedic Surgery, National Clinical Research Center for Geriatrics, West China Hospital of Sichuan University, Chengdu, Sichuan Province, China;2. Department of Integrated Traditional Chinese and Western Medicine, West China Hospital of Sichuan University, Chengdu, Sichuan Province, China;3. Core Facility of West China Hospital, Sichuan University, Chengdu, Sichuan Province, China;4. Laboratory of Endocrinology and Metabolism, Department of Department of Endocrinology and Metabolism, West China Hospital of Sichuan University, Chengdu, Sichuan Province, China;1. Department of Orthopedics, The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai 519000, Guangdong, China;2. Guangdong Provincial Key Laboratory of Biomedical Imaging, The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai 519000, Guangdong, China;3. Department of Orthopedics, The Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai 264000, Shandong, China;2. Department of Orthopaedics and Traumatology, The University of Hong Kong, Pokfulam, Hong Kong;3. Department of Anatomy, The University of Hong Kong, Pokfulam, Hong Kong;4. Heart, Brain, Hormone and Healthy Aging Center, The University of Hong Kong, Pokfulam, Hong Kong;6. State Key Laboratory for Pharmaceutical Biotechnology, LKS Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong;5. Georgetown University Medical Center, Washington, DC 20057, USA |
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| Abstract: | Endothelin (ET) receptors on chondrocytes were demonstrated using cultured rabbit costal chondrocytes. After crosslinking the receptors on the cells with 125 I-ET-1, two major bands of 43 kDa and 46 kDa were separated by SDS-PAGE. Scatchard analysis demonstrated two classes of ET receptors with Kd values of 1 × 10-10 M and 5 × 10-9 M. The numbers of high- and low- affinity receptors were 1 × 104 and 2 × 105 per cell, respectively. The binding of ET-1 to chondrocytes was increased by treatment with PTH, DBcAMP, TGF-β1, IL-1β, RA and EGF. ET-1 stimulated DNA synthesis in cultured rabbit chondrocytes. ET-1 also stimulated calcium incorporation through the cell membrane of chondrocytes. These findings indicate that ET-1 has a physiological effect on chondrocytes via its receptors on the cells. |
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