机构：[1]The Division of Pulmonary and Critical Care Medicine, Guangzhou Institute of Respiratory Health, National Clinical Research Center for Respiratory Disease, State Key Laboratory of Respiratory Disease, National Center for Respiratory Medicine, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, China,[2]Department of Intensive Care Unit, the Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, China,[3]Department of Acupuncture, Guangdong Provincial Hospital of Traditional Chinese Medicine, Guangzhou University of Traditional Chinese Medicine, Guangzhou, Guangdong, China,大德路总院针灸科大德路总院针灸科广东省中医院[4]Respiratory Medicine, Hunan Provincial People's Hospital, Changsha, Hunan, China,[5]GMU-GIBH Joint School of Life Sciences, Guangzhou Medical University, Guangzhou, Guangdong,China
Although both calcium sensing receptor (CaSR) and canonical transient receptor potential (TRPC) proteins contribute to chronic hypoxia (CH)-induced pulmonary arterial smooth muscle cells (PASMCs) proliferation, the relationship between CaSR and TRPC in hypoxic PASMCs proliferation remains poorly understood. We hypothesized that CH promotes PASMCs proliferation through CaSR-TRPC pathway. Rat PASMCs were isolated and treated with CH. Cell proliferation was assessed by cell counting, CCK-8 assay, and EdU incorporation. CaSR and TRPC expressions were determined by qPCR and Western blotting. Store-operated Ca2+ entry (SOCE) was assessed by extracellular Ca2+ restoration. In PASMCs, CH enhanced the cell number, cell viability and DNA synthesis, which is accompanied by upregulated expression of CaSR, TRPC1 and TRPC6. Negative CaSR modulators (NPS2143 and NPS2390) inhibited, whereas positive modulators (spermine, R568) enhanced, the CH-induced increases in cell number, cell viability and DNA synthesis in PASMCs. Knockdown of CaSR by siRNA inhibited the CH-induced upregulation of TRPC1 and TRPC6 and enhancement of SOCE and attenuated the CH-induced enhancements of cell number, cell viability and DNA synthesis in PASMCs. However, neither siTRPC1 nor siTRPC6 had an effect on the CH-induced CaSR upregulation, although both significantly attenuated the CH-induced enhancements of cell number, cell viability and DNA synthesis in PASMCs. These results demonstrate that upregulated CaSR-TRPC1/6 pathway mediating PASMCs proliferation is an important pathogenic mechanism under hypoxic conditions.
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基金：
the Local Innovative and Research Teams Project of Guangdong Pearl River Talents Program (2017BT01S155), the Natural Science Foundation of Guangdong
Province (2021A1515011024, 2017A030313683, 2017A030310419), National key Research and
Development program of China (2016YFC1304100, 2016YFC1304104), National Natural Science
Foundation of China (81570045), the Science and technology program of Guangzhou
(201504010018), the Training Program for Academic Backbone of High Level Universities of
Guangzhou Medical University (2017210), the Grant of the First Affiliated Hospital of
Guangzhou Medical University (201619), and the Grant of State Key Laboratory of Respiratory
Disease (SKLRD2016ZJ013). Dr. Peng was supported by the Scholarship of Guangzhou Medical
University and the First Affiliated Hospital of Guangzhou Medical University.
第一作者机构：[1]The Division of Pulmonary and Critical Care Medicine, Guangzhou Institute of Respiratory Health, National Clinical Research Center for Respiratory Disease, State Key Laboratory of Respiratory Disease, National Center for Respiratory Medicine, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, China,[2]Department of Intensive Care Unit, the Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, China,
共同第一作者：
通讯作者：
通讯机构：[1]The Division of Pulmonary and Critical Care Medicine, Guangzhou Institute of Respiratory Health, National Clinical Research Center for Respiratory Disease, State Key Laboratory of Respiratory Disease, National Center for Respiratory Medicine, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, China,[*1]The Division of Pulmonary and Critical Care Medicine, Guangzhou Institute of Respiratory Health, National Clinical Research Center for Respiratory Disease, State Key Laboratory of Respiratory Disease, National Center for Respiratory Medicine, The First Affiliated Hospital of Guangzhou Medical University 151 Yanjiang Road, Guangzhou, Guangdong 510120, China
