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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| Format: | Article |
| Language: | zho |
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Editorial Office of Journal of Third Military Medical University
2021-11-01
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| Series: | Di-san junyi daxue xuebao |
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| Online Access: | http://aammt.tmmu.edu.cn/Upload/rhtml/202107145.htm |
| _version_ | 1828960373820096512 |
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| 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 |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-14T09:26:05Z |
| format | Article |
| id | doaj.art-1bab4b0f9abb4c3d9f7a237e4c2c31b5 |
| institution | Directory Open Access Journal |
| issn | 1000-5404 |
| language | zho |
| last_indexed | 2024-12-14T09:26:05Z |
| publishDate | 2021-11-01 |
| publisher | Editorial Office of Journal of Third Military Medical University |
| record_format | Article |
| series | Di-san junyi daxue xuebao |
| spelling | doaj.art-1bab4b0f9abb4c3d9f7a237e4c2c31b52022-12-21T23:08:11ZzhoEditorial Office of Journal of Third Military Medical UniversityDi-san junyi daxue xuebao1000-54042021-11-0143212350235710.16016/j.1000-5404.202107145TLR9-JNK signaling pathway regulates hypoxia-induced proliferation of rat pulmonary artery smooth muscle cellsWU Qijian0WU Qijian1 ZHANG Erlong2 ZHANG Erlong3SUN Binda4SUN Binda5Department of Medicine and Equipment for High Altitude Region, Army Medical University (Third Military Medical University), Chongqing, 400038, ChinaKey Laboratory of Extreme Environmental Medicine of Ministry of Education, Army Medical University (Third Military Medical University), Chongqing, 400038, ChinaDepartment of Medicine and Equipment for High Altitude Region, Army Medical University (Third Military Medical University), Chongqing, 400038, ChinaKey Laboratory of Extreme Environmental Medicine of Ministry of Education, Army Medical University (Third Military Medical University), Chongqing, 400038, ChinaDepartment of Medicine and Equipment for High Altitude Region, Army Medical University (Third Military Medical University), Chongqing, 400038, ChinaKey Laboratory of Extreme Environmental Medicine of Ministry of Education, Army Medical University (Third Military Medical University), Chongqing, 400038, ChinaObjective 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.http://aammt.tmmu.edu.cn/Upload/rhtml/202107145.htmtoll-like receptor 9c-jun n-terminal kinaserat pulmonary artery smooth muscle cellshypoxiacell proliferation |
| spellingShingle | 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 Di-san junyi daxue xuebao toll-like receptor 9 c-jun n-terminal kinase rat pulmonary artery smooth muscle cells hypoxia cell proliferation |
| title | 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_short | 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 |
| topic | toll-like receptor 9 c-jun n-terminal kinase rat pulmonary artery smooth muscle cells hypoxia cell proliferation |
| url | http://aammt.tmmu.edu.cn/Upload/rhtml/202107145.htm |
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