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

Qijian WU; Erlong ZHANG; Binda SUN; Gang XU; Jian HUANG; Yuqi GAO

doi:10.16016/j.1000-5404.202107145

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

Basic Medicine | 更新时间：2026-01-09

- TLR9-JNK signaling pathway regulates hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells
- Journal of Third Military Medical University Vol. 43, Issue 21, Pages: 2350-2357(2021)
- 作者机构：
  
  1.400038 重庆，陆军军医大学(第三军医大学)高原军事医学系：高原特需药品与器材研究室
  2.400038 重庆，陆军军医大学(第三军医大学)高原军事医学系：高原生理学与病理学教研室
  3.400038 重庆，陆军军医大学(第三军医大学)高原军事医学系：极端环境医学教育部重点实验室
- 作者简介：
- 基金信息：
  
  the Key Program of National Natural Science Foundation of China(81830062)
- DOI：10.16016/j.1000-5404.202107145
  CLC： R332.121;R329.28;R363.22
- Received：23 July 2021，
  
  Revised：2021-09-25，
  
  Published：15 November 2021
- 稿件说明：
移动端阅览
Qijian WU, Erlong ZHANG, Binda SUN, et al. TLR9-JNK signaling pathway regulates hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells[J]. Journal of Third Military Medical University, 2021, 43(21): 2350-2357.
DOI：

Qijian WU, Erlong ZHANG, Binda SUN, et al. TLR9-JNK signaling pathway regulates hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells[J]. Journal of Third Military Medical University, 2021, 43(21): 2350-2357. DOI： 10.16016/j.1000-5404.202107145.

摘要

目的

研究Toll样受体-9(toll-like receptor 9，TLR9)在缺氧诱导大鼠肺动脉平滑肌细胞(rat pulmonary artery smooth muscle cells，RPASMCs)增殖中的作用及机制。

方法

采用MTT、Western blot、PCR法检测RPASMCs的相对增殖水平、蛋白表达水平、mRNA转录水平。①3% O

，36 h处理RPASMCs，构建缺氧诱导RPASMCs增殖的细胞模型，检测细胞相对增殖水平、TLR9、磷酸化c-jun氨基末端激酶(phosphorylated c-jun N-terminal kinase，p-JNK)蛋白表达水平。②缺氧条件下分别使用小干扰RNA序列(small interfering RNA sequence，siRNA)或20 μmol/L TLR9激动剂ODN 1826敲低或活化TLR9，检测RPASMCs的细胞增殖水平和c-jun氨基末端激酶(c-jun N-terminal kinase，JNK)磷酸化水平。③缺氧时使用5 μmol/L JNK抑制剂SP600125处理RPASMCs，检测缺氧条件下细胞相对增殖水平。④缺氧时复合使用20 μmol/L TLR9激动剂ODN 1826和5 μmol/L JNK抑制剂SP600125处理RPASMCs，检测RPASMCs的JNK磷酸化水平和相对增殖水平。

结果

缺氧组RPASMCs的相对增殖水平[(1.30±0.07)，

0.01

]

以及TLR9蛋白水平[(1.70±0.22)，

0.05

]

和JNK磷酸化[(1.83±0.18)，

0.01

]

都显著高于常氧组。敲低TLR9显著抑制缺氧诱导的RPASMC增殖[(0.64±0.09)，

0.01

]

并减少JNK磷酸化[(0.49±0.01)，

0.001

]

。TLR9激动剂显著促进RPASMCs增殖[(2.01±0.08)，

0.001

]

且增加JNK的磷酸化[(2.11±0.33)，

0.01

]

。JNK抑制剂抑制RPASMCs增殖[(0.70±0.06)，

0.05

]

。与缺氧+TLR9激动剂组相比，缺氧+TLR9激动剂+JNK抑制剂组RPASMCs JNK磷酸化[(0.82±0.14)

(1.59±0.30)，

0.001

]

和相对增殖水平[(1.08±0.05)

(1.35± 0.02)，

0.001

]

都显著降低。

结论

缺氧时TLR9通过激活JNK通路促进大鼠肺动脉平滑肌细胞增殖。

Abstract

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% O

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

0.01) and protein levels of TLR9 (1.70±0.22

0.05) and p-JNK (1.83±0.18

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

0.01) and reduced the JNK phosphorylation (0.49±0.01

0.001). TLR9 agonist

ODN 1826

significantly promoted the proliferation of RPASMCs (2.01±0.08

0.001) and increased JNK phosphorylation (2.11±0.33

0.01). JNK inhibitor SP600125 inhibited the proliferation (0.70±0.06

0.05). Compared with hypoxia+TLR9 agonist group

the level of p-JNK (0.82±0.14

1.59±0.30

0.001) and cell proliferation (1.08±0.05

1.35±0.02

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.

关键词

Keywords

references

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