LPS inhibits arginase-1 expression in macrophages by up-regulating activity of cytochrome P450 1A1 epoxidase

Lixing TIAN; Junyu ZHU; Linghui WAN; Huaping LIANG

doi:10.16016/j.2097-0927.202208218

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LPS inhibits arginase-1 expression in macrophages by up-regulating activity of cytochrome P450 1A1 epoxidase

Basic Medicine | 更新时间：2026-01-09

- LPS inhibits arginase-1 expression in macrophages by up-regulating activity of cytochrome P450 1A1 epoxidase
- Journal of Army Medical University Vol. 45, Issue 17, Pages: 1790-1796(2023)
- 作者机构：
  
  1.831200 新疆呼图壁，陆军军医大学(第三军医大学)边防卫勤训练大队军事预防医学教研室
  2.400042 重庆，陆军军医大学(第三军医大学)陆军特色医学中心战伤感染与特需药品研究室
  3.831200 新疆呼图壁，陆军军医大学(第三军医大学)边防卫勤训练大队基础医学教研室
- 作者简介：
- 基金信息：
  
  the Special Project of Science and Technology Innovation Capacity Promotion of Army Medical University(2021XQN11);the General Program of Natural Science Foundation of Chongqing(cstc2021jcyj-msxmX0234)
- DOI：10.16016/j.2097-0927.202208218
  CLC： R345;R364.5;R392.3
- Received：30 August 2022，
  
  Revised：2023-07-31，
  
  Published：15 September 2023
- 稿件说明：
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Lixing TIAN, Junyu ZHU, Linghui WAN, et al. LPS inhibits arginase-1 expression in macrophages by up-regulating activity of cytochrome P450 1A1 epoxidase[J]. Journal of Army Medical University, 2023, 45(17): 1790-1796.
DOI：

Lixing TIAN, Junyu ZHU, Linghui WAN, et al. LPS inhibits arginase-1 expression in macrophages by up-regulating activity of cytochrome P450 1A1 epoxidase[J]. Journal of Army Medical University, 2023, 45(17): 1790-1796. DOI： 10.16016/j.2097-0927.202208218.

摘要

目的

探索细胞色素P450 1A1(cytochrome P450 1A1，CYP1A1)对脂多糖(lipopolysaccharide，LPS)诱导时巨噬细胞中精氨酸酶-1(Arginase-1，Arg-1)的调控作用及机制。

方法

使用LPS刺激小鼠单核巨噬细胞细胞系RAW264.7(RAW)，分为0 h组、12 h组、24 h组(每组设3复孔

=3)，采用实时荧光定量PCR(RT-qPCR)及蛋白免疫印迹实验(Western blot)检测CYP1A1、Arg-1 mRNA及蛋白表达；构建阴性对照和高表达、敲除CYP1A1的RAW(NC/RAW、CYP1A1/RAW、CYP1A1 KO/RAW)细胞系以及突变CYP1A1羟化酶活性的RAW(CYP1A1m/RAW)细胞系，分为NC/RAW+LPS组、CYP1A1 KO/RAW+LPS组、CYP1A1/RAW+LPS组、CYP1A1m/RAW+LPS组，使用LPS刺激后检测Arg-1 mRNA、蛋白表达及信号转导和转录活化因子6(signal transducer and activators of transcription-6，STAT6)活化水平；使用STAT6抑制剂AS1517499作用CYP1A1敲除RAW细胞，设立NC/RAW+PBS+LPS组、NC/RAW+AS1517499+LPS组、CYP1A1 KO/RAW+PBS+LPS组、CYP1A1 KO/RAW+AS1517499+LPS组，检测Arg-1 mRNA、蛋白表达；使用CYP1A1环氧化

酶活性抑制剂甲二氢愈创木酸(Nordihydroguaiaretic Acid，NDGA)及12(S)-羟基二十碳四烯酸[12(S)-HETE

]

作用RAW细胞，设立RAW+PBS+LPS组、RAW+12(S)-HETE+LPS组、RAW+NDGA+LPS组，检测Arg-1 mRNA、蛋白表达及STAT6活化水平。

结果

LPS可诱导RAW中的CYP1A1及Arg-1呈错峰高表达(

＜0.05)；CYP1A1高表达可抑制LPS诱导时RAW中Arg-1表达(

＜0.05)及STAT6活化，敲除则可增强上述效应(

＜0.05)，而STAT6抑制剂AS1517499可消除CYP1A1敲除带来的增强效应(

＜0.05)；突变CYP1A1的羟化酶活性或使用其代谢产物12(S)-HETE刺激均不影响Arg-1表达(

＞0.05)，而使用CYP1A1环氧化酶抑制剂NDGA可增强Arg-1表达(

＜0.05)及STAT6活化。

结论

CYP1A1可通过抑制LPS诱导时巨噬细胞中STAT6活化水平以降低Arg-1表达，但此效应与CYP1A1羟化酶活性及其致炎产物12(S)-HETE无关，而可能与CYP1A1环氧化酶活性有关。

Abstract

Objective

To explore the regulative effect and mechanism of cytochrome P450 1A1 (CYP1A1) on regulating lipopolysaccharide (LPS)-induced arginase-1 (Arg-1) expression in macrophages.

Methods

After mouse macrophage-like cell line RAW264.7 (RAW) was treated with 10 mg/L LPS for 0

12 and 24 h

respectively (

=3 for each time point)

the expression of CYP1A1 and Arg-1 at mRNA and protein levels were measured by RT-PCR and Western blotting. Negative control RAW(NC/RAW) cells

CYP1A1-overexpression RAW (CYP1A1/RAW) cells

CYP1A1-knockout RAW (CYP1A1 KO/RAW) cells and CYP1A1 hydroxylase-activity mutant RAW (CYP1A1m/RAW) cells were constructed

respectively

and then the cells were treated with LPS and further detected with RT-PCR and Western blotting for the expression and activation levels of Arg-1 and signal transducer and activators of transcription-6 (STAT6). Then STAT 6 inhibitor AS1517499 was added to treat the CYP1A1 KO/RAW cells and NC/RAW cells

and the mRNA and protein expression of Arg-1 was measured again after LPS or PBS treatment. CYP1A1 epoxidase inhibitor

nordihydroguaiaretic acid (NDGA) and 12 (S)-HETE were introduced to treat normal RAW cells

the mRNA and protein levels of Arg-1 an

d activity of STAT6 were detected after LPS or PBS treatment.

Results

LPS-induced CYP1A1 and Arg-1 were highly expressed in staggering peaks in RAW cells (

0.05). Overexpression of CYP1A1 inhibited LPS-induced Arg-1 expression and STAT6 activation in RAW cells

while CYP1A1 knockout enhanced above effects (

0.05). STAT6 inhibitor AS1517499 abolished the enhancement of CYP1A1 knockout (

0.05). CYP1A1 hydroxylase-activity mutation or 12(S)-HETE simulation had no effect on the expression of Arg-1 (

0.05)

while CYP1A1 epoxidase-activity inhibitor NDGA enhanced Arg-1 expression and STAT6 activation (

0.05).

Conclusion

CYP1A1 can inhibit Arg-1 expression by reducing STAT6 activation in LPS-induced macrophages

and CYP1A1 hydroxylase-activity or its proinflammatory metabolism 12(S)-HETE has no effect on this regulation

while CYP1A1 epoxidase-activity may be involved in this effect.

关键词

Keywords

references

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