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5-HT Promote Epithelial to Mesenchymal Transition Via 5-HT(2A/2B) Receptors in 16-HBE
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A3818 - 5-HT Promote Epithelial to Mesenchymal Transition Via 5-HT(2A/2B) Receptors in 16-HBE
Author Block: T. Yang1, H. Wang2, Y. Shen2, C. Wan2, F. Wen2; 1West China Hospital of Si Chuan University, Cheng Du, China, 2West China Hospital of Si Chuan University, Chengdu, China.
Rationale: Epithelial to mesenchymal transition (EMT) had been found to be involved in the pathogenesis of idiopathic pulmonary fibrosis. Serotonin(5-hydroxytryptamine; 5-HT) combined with its two receptors, 5-HT2A and 5-HT2B, is known to increase proliferation and collagen synthesis by fibroblasts, thus, plays a role in bleomycin induced lung fibrosis. While the role of 5-HT and 5-HT (2A/2B) receptors in epithelial to mesenchymal transition is still unknown. Methods: Immunohistochemistry assays were utilized to analyze the expression and localization of 5-HT (2A/2B) receptors in two airway epithelial cells (16-HBE and NCI-H292). 16-HBE cells were incubated with vehicle or 5-HT (8, 16 and 24uM). Then 16-HBE cells were pre-incubated with Ketanserin (5-HT2A receptors inhibitor) and RS-127445 (5-HT2B receptors inhibitor), then exposure to 16uM 5-HT. Real time-PCR was used to analysis the mRNA level of CTGF, PDGF and collagen-1a.Results: Immunohistochemistry assays results showed that the 5-HT (2A/2B)receptors were positive both in the membrane and cytoplasm of the human bronchial epithelial cell line (16-HBE), not in NCI-H292. So, we explore the effect of 5-HT on epithelial to mesenchymal transition in 16-HBE. The mRNA level of EMT related genes increased after exposure to 5-HT at 16uM, including CTGF(Connective Tissue Growth Factor),PDGF(platelet derived growth factor), and collagen-1a. RS-127445 inhibited the mRNA level of CTGF, PDGF and collagen-1a induced by 5-HT. Ketanserin inhibited the mRNA level of CTGF. RS-127445. While RS-127445 combined with Ketanserin also inhibited the mRNA level of collagen-1a. Conclusions: Our results suggest that 5-HT may induced epithelial to mesenchymal transition partially by increasing the mRNA levels of EMT related genes, and which was mediated by the 5-HT (2A/2B) receptors.
Author Block: T. Yang1, H. Wang2, Y. Shen2, C. Wan2, F. Wen2; 1West China Hospital of Si Chuan University, Cheng Du, China, 2West China Hospital of Si Chuan University, Chengdu, China.
Rationale: Epithelial to mesenchymal transition (EMT) had been found to be involved in the pathogenesis of idiopathic pulmonary fibrosis. Serotonin(5-hydroxytryptamine; 5-HT) combined with its two receptors, 5-HT2A and 5-HT2B, is known to increase proliferation and collagen synthesis by fibroblasts, thus, plays a role in bleomycin induced lung fibrosis. While the role of 5-HT and 5-HT (2A/2B) receptors in epithelial to mesenchymal transition is still unknown. Methods: Immunohistochemistry assays were utilized to analyze the expression and localization of 5-HT (2A/2B) receptors in two airway epithelial cells (16-HBE and NCI-H292). 16-HBE cells were incubated with vehicle or 5-HT (8, 16 and 24uM). Then 16-HBE cells were pre-incubated with Ketanserin (5-HT2A receptors inhibitor) and RS-127445 (5-HT2B receptors inhibitor), then exposure to 16uM 5-HT. Real time-PCR was used to analysis the mRNA level of CTGF, PDGF and collagen-1a.Results: Immunohistochemistry assays results showed that the 5-HT (2A/2B)receptors were positive both in the membrane and cytoplasm of the human bronchial epithelial cell line (16-HBE), not in NCI-H292. So, we explore the effect of 5-HT on epithelial to mesenchymal transition in 16-HBE. The mRNA level of EMT related genes increased after exposure to 5-HT at 16uM, including CTGF(Connective Tissue Growth Factor),PDGF(platelet derived growth factor), and collagen-1a. RS-127445 inhibited the mRNA level of CTGF, PDGF and collagen-1a induced by 5-HT. Ketanserin inhibited the mRNA level of CTGF. RS-127445. While RS-127445 combined with Ketanserin also inhibited the mRNA level of collagen-1a. Conclusions: Our results suggest that 5-HT may induced epithelial to mesenchymal transition partially by increasing the mRNA levels of EMT related genes, and which was mediated by the 5-HT (2A/2B) receptors.
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