Transient Receptor Potential Ankyrin 1 (TRPA1) Mediates Lipopolysaccharide (LPS)-Induced Inflammatory Responses in Primary Human Osteoarthritic Fibroblast-Like Synoviocytes
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Transient receptor potential ankyrin 1 (TRPA1) is a membrane-associated cation channel, widely expressed in neuronal and non-neuronal cells. Recently, emerging evidences suggested the crucial role of TRPA1 in the disease progression of osteoarthritis (OA). Therefore, we aimed to investigate whether TRPA1 mediate lipopolysaccharide (LPS)-induced inflammatory responses in primary human OA fibroblast-like synoviocytes (OA-FLS). The expression of TRPA1 in LPS-treated OA-FLS was assessed by polymerase chain reaction (PCR) and western blot (WB), and the functionality of TRPA1 channel by Ca2+ influx measurements. Meanwhile, production of interleukin (IL)-1β, tumor necrosis factor (TNF)-α, IL-6, matrix metalloproteinase (MMP)-1, and MMP-3 in LPS-treated cells was measured by immunoassay. Histological observation after inhibition of TRPA1 was also performed in rats with LPS-induced inflammatory arthritis. After being induced by LPS, the gene and protein expression of TRPA1 was increased in the time-dependent or dose-dependent manner. Meanwhile, Ca2+ influx mediated by TRPA1 in human OA-FLS was also enhanced. In addition, pharmacological inhibition and gene silencing of TRPA1 downregulated the production of IL-1β, TNF-α, IL-6, MMP-1, and MMP-3 in LPS-treated FLS. Finally, synovial inflammation and cartilage degeneration were also reduced by the TRPA1 antagonist. We found the LPS caused the increased functional expression of TRPA1, the activation of which involved in LPS-reduced inflammatory responses in primary human OA-FLS, and the inhibition of TRPA1 produces protective effect in LPS-induced arthritis.
KEY WORDStransient receptor potential ankyrin 1 lipopolysaccharide osteoarthritis fibroblast-like synoviocytes
The authors wish to express their gratitude to all staffs in the medical research center of first college of clinical medicine, the Nanjing University of Chinese Medicine, Nanjing, China.
SongJiang Yin and Peimin Wang conceived the study and drafted the manuscript. Li Zhang and Linrui Zhao designed and performed the animal experiments; Xiaochen Li and Yancheng Xiao performed cell experiments. All authors read and approved the final manuscript. All authors approved the final version and are accountable for all aspects of the manuscript.
This research was supported by the National Natural Science Foundation of China (81573993), the National Natural Science Foundation of China (81774334), the Natural Science Foundation of Jiangsu Province(BK20171513) and Natural Science Foundation of Jiangsu Province (BK20151598).
Compliance with Ethical Standards
Ethics Approval and Consent to Participate
All animal protocols were approved (reference number: ACU170701) by the Animal Care and Use Committee of the Nanjing University of Chinese Medicine. All experiments were conducted in accordance with the National Institutes of Health Guidelines for the Care and Use of Laboratory Animals.
Synovial tissues in knee OA patients obtained from knee joint replacement surgery were removed with patients’ consent. The patients in this study met the American College of Rheumatology classification criteria for OA and the study was approved (reference number: 2015NL-068-02) by the Ethics Committee of Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, China.
Consent for Publication
Data was de-identified, after collection, and therefore, there was no consent required from individual patients, according to the ethical approval.
The authors declare that there is no conflict of interests.
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