TLR9-JNK signaling pathway regulates hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells

Objective To investigate the role and mechanism of Toll-like receptor 9 (TLR9) in hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells (RPASMCs). Methods MTT assay, Western blotting and PCR were adopted to measure cell proliferation level, protein and mRNA levels of following mo...

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Autores principales:	WU Qijian, ZHANG Erlong, SUN Binda
Formato:	article
Lenguaje:	ZH
Publicado:	Editorial Office of Journal of Third Military Medical University 2021
Materias:	toll-like receptor 9 c-jun n-terminal kinase rat pulmonary artery smooth muscle cells hypoxia cell proliferation Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/1bab4b0f9abb4c3d9f7a237e4c2c31b5
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spelling	oai:doaj.org-article:1bab4b0f9abb4c3d9f7a237e4c2c31b52021-11-12T04:58:11ZTLR9-JNK signaling pathway regulates hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells10.16016/j.1000-5404.2021071451000-5404https://doaj.org/article/1bab4b0f9abb4c3d9f7a237e4c2c31b52021-11-01T00:00:00Zhttp://aammt.tmmu.edu.cn/Upload/rhtml/202107145.htmhttps://doaj.org/toc/1000-5404Objective To investigate the role and mechanism of Toll-like receptor 9 (TLR9) in hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells (RPASMCs). Methods MTT assay, Western blotting and PCR were adopted to measure cell proliferation level, protein and mRNA levels of following molecules in RPASMCs. ① After RPASMCs were treated with 3% O2 for 36 h to establish a hypoxic model, the cell proliferation and expression levels of TLR9 and phosphorylated c-jun N-terminal kinase(p-JNK) were detected at the same time. ② After TLR9 was knocked down using small interfering RNA sequence (siRNA) or activated by 20 μmol/L TLR9 agonist, ODN 1826, under hypoxia, the proliferation and p-JNK level were examined. ③RPASMCs were treated with 5 μmol/L JNK inhibitor SP600125 under hypoxia, and the proliferation was assayed. ④ Under hypoxia, 20 μmol/L ODN 1826 combined with 5 μmol/L SP600125 were administered to RPASMCs, and the proliferation and level of p-JNK were measured. Results The proliferation of RPASMCs (1.30±0.07, P < 0.01) and protein levels of TLR9 (1.70±0.22, P < 0.05) and p-JNK (1.83±0.18, P < 0.01) were significantly higher in the hypoxia group than the normoxia group. Knock-down of TLR9 significantly inhibited hypoxia-induced proliferation of RPASMC (0.64±0.09, P < 0.01) and reduced the JNK phosphorylation (0.49±0.01, P < 0.001). TLR9 agonist, ODN 1826, significantly promoted the proliferation of RPASMCs (2.01±0.08, P < 0.001) and increased JNK phosphorylation (2.11±0.33, P < 0.01). JNK inhibitor SP600125 inhibited the proliferation (0.70±0.06, P < 0.05). Compared with hypoxia+TLR9 agonist group, the level of p-JNK (0.82±0.14 vs 1.59±0.30, P < 0.001) and cell proliferation (1.08±0.05 vs 1.35±0.02, P < 0.001) were both decreased significantly in hypoxia+TLR9 agonist+JNK inhibitor group. Conclusion TLR9 promotes the proliferation of RPASMCs by activating the JNK pathway under hypoxia.WU QijianWU Qijian ZHANG Erlong ZHANG ErlongSUN BindaSUN BindaEditorial Office of Journal of Third Military Medical Universityarticletoll-like receptor 9c-jun n-terminal kinaserat pulmonary artery smooth muscle cellshypoxiacell proliferationMedicine (General)R5-920ZHDi-san junyi daxue xuebao, Vol 43, Iss 21, Pp 2350-2357 (2021)
institution	DOAJ
collection	DOAJ
language	ZH
topic	toll-like receptor 9 c-jun n-terminal kinase rat pulmonary artery smooth muscle cells hypoxia cell proliferation Medicine (General) R5-920
spellingShingle	toll-like receptor 9 c-jun n-terminal kinase rat pulmonary artery smooth muscle cells hypoxia cell proliferation Medicine (General) R5-920 WU Qijian WU Qijian ZHANG Erlong ZHANG Erlong SUN Binda SUN Binda TLR9-JNK signaling pathway regulates hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells
description	Objective To investigate the role and mechanism of Toll-like receptor 9 (TLR9) in hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells (RPASMCs). Methods MTT assay, Western blotting and PCR were adopted to measure cell proliferation level, protein and mRNA levels of following molecules in RPASMCs. ① After RPASMCs were treated with 3% O2 for 36 h to establish a hypoxic model, the cell proliferation and expression levels of TLR9 and phosphorylated c-jun N-terminal kinase(p-JNK) were detected at the same time. ② After TLR9 was knocked down using small interfering RNA sequence (siRNA) or activated by 20 μmol/L TLR9 agonist, ODN 1826, under hypoxia, the proliferation and p-JNK level were examined. ③RPASMCs were treated with 5 μmol/L JNK inhibitor SP600125 under hypoxia, and the proliferation was assayed. ④ Under hypoxia, 20 μmol/L ODN 1826 combined with 5 μmol/L SP600125 were administered to RPASMCs, and the proliferation and level of p-JNK were measured. Results The proliferation of RPASMCs (1.30±0.07, P < 0.01) and protein levels of TLR9 (1.70±0.22, P < 0.05) and p-JNK (1.83±0.18, P < 0.01) were significantly higher in the hypoxia group than the normoxia group. Knock-down of TLR9 significantly inhibited hypoxia-induced proliferation of RPASMC (0.64±0.09, P < 0.01) and reduced the JNK phosphorylation (0.49±0.01, P < 0.001). TLR9 agonist, ODN 1826, significantly promoted the proliferation of RPASMCs (2.01±0.08, P < 0.001) and increased JNK phosphorylation (2.11±0.33, P < 0.01). JNK inhibitor SP600125 inhibited the proliferation (0.70±0.06, P < 0.05). Compared with hypoxia+TLR9 agonist group, the level of p-JNK (0.82±0.14 vs 1.59±0.30, P < 0.001) and cell proliferation (1.08±0.05 vs 1.35±0.02, P < 0.001) were both decreased significantly in hypoxia+TLR9 agonist+JNK inhibitor group. Conclusion TLR9 promotes the proliferation of RPASMCs by activating the JNK pathway under hypoxia.
format	article
author	WU Qijian WU Qijian ZHANG Erlong ZHANG Erlong SUN Binda SUN Binda
author_facet	WU Qijian WU Qijian ZHANG Erlong ZHANG Erlong SUN Binda SUN Binda
author_sort	WU Qijian
title	TLR9-JNK signaling pathway regulates hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells
title_short	TLR9-JNK signaling pathway regulates hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells
title_full	TLR9-JNK signaling pathway regulates hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells
title_fullStr	TLR9-JNK signaling pathway regulates hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells
title_full_unstemmed	TLR9-JNK signaling pathway regulates hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells
title_sort	tlr9-jnk signaling pathway regulates hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells
publisher	Editorial Office of Journal of Third Military Medical University
publishDate	2021
url	https://doaj.org/article/1bab4b0f9abb4c3d9f7a237e4c2c31b5
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TLR9-JNK signaling pathway regulates hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells

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