LOC339524 gene mediates lipopolysaccharide-induced expression of inflammatory cytokines in mouse microglia bv2 cell line

Xianglong TANG; Zonghong LONG; Guangyou DUAN; Xiaoying ZHOU; Huifang CHEN; Hong LI

doi:10.16016/j.1000-5404.201810166

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LOC339524 gene mediates lipopolysaccharide-induced expression of inflammatory cytokines in mouse microglia bv2 cell line

Basic Medicine | 更新时间：2026-01-09

- LOC339524 gene mediates lipopolysaccharide-induced expression of inflammatory cytokines in mouse microglia bv2 cell line
- Journal of Third Military Medical University Vol. 41, Issue 4, Pages: 289-295(2019)
- 作者机构：
  
  400037 重庆, 陆军军医大学(第三军医大学)第二附属医院麻醉科
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81571870)
- DOI：10.16016/j.1000-5404.201810166
  CLC： R392-33;R392.11;R392.13
- Received：25 October 2018，
  
  Revised：2018-12-07，
  
  Published：28 February 2019
- 稿件说明：
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Xianglong TANG, Zonghong LONG, Guangyou DUAN, et al. LOC339524 gene mediates lipopolysaccharide-induced expression of inflammatory cytokines in mouse microglia bv2 cell line[J]. Journal of Third Military Medical University, 2019, 41(4): 289-295.
DOI：

Xianglong TANG, Zonghong LONG, Guangyou DUAN, et al. LOC339524 gene mediates lipopolysaccharide-induced expression of inflammatory cytokines in mouse microglia bv2 cell line[J]. Journal of Third Military Medical University, 2019, 41(4): 289-295. DOI： 10.16016/j.1000-5404.201810166.

摘要

目的

研究LOC339524基因对脂多糖(lipopolysaccharide，LPS)诱导的小鼠小胶质bv2细胞系炎症因子表达的影响。

方法

100 ng/mL LPS处理bv2细胞3、6、9、12 h用于摸索最佳炎症反应时间；10、100、1 000、10 000 ng/mL LPS处理bv2细胞和100 ng/mL LPS处理bv2细胞3、6、9、12 h用于研究LOC339524蛋白表达变化；siRNA沉默LOC339524和慢病毒转染构建过表达LOC339524的细胞株后加LPS处理用于检测炎症因子变化；qPCR检测TNF-α、IL-6和LOC339524 mRNA水平变化，Western blot检测TNF-α、IL-6和LOC339524蛋白水平变化。

结果

与0 h相比，LPS处理不同时间均可使TNF-α、IL-6 mRNA和蛋白表达增加(

0.05)，12 h两者表达一致，选择12 h进行后续炎症反应研究；与0 ng/mL LPS处理组相比，不同浓度LPS均可诱导LOC339524蛋白的表达增加(

0.05)，100 ng/mL LPS处理后呈时间依赖性增加(

0.05)。LPS处理12 h后，siRNA组较无关片段组，TNF-α和IL-6 mRNA和蛋白表达显著上调(

0.05)，反之，过表达组较正常组TNF-α和IL-6 mRNA和蛋白表达显著下调(

0.05)。

结论

LOC339524基因介导了脂多糖诱导bv2细胞的炎症反应。

Abstract

Objective

To study the role of LOC339524 gene in mediating the expression of inflammatory cytokines in mouse microglia bv2 cell line induced by lipopolysaccharide (LPS).

Methods

Mouse microglia bv2 cells were treated with 100 ng/mL LPS for 3

9 or 12 h to determine the optimal time for observing the inflammatory response. The cells were then treated with 10

100

1 000

and 10 000 ng/mL LPS or with 100 ng/mL LPS for 3

and 12 h

and the changes in the expression of LOC339524 protein was investigated. The changes in the expression of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in response to LPS challenge were detected in bv2 cells with siRNA-mediated LOC339524 gene silencing or lentivirus-mediated LOC339524 overexpression using qPCR and Western blotting.

Results

Compared with blank control

LPS treatment increased TNF-α

IL-6 mRNA and protein expression at different times (

0.05)

and the expression was consistent at 12 hours. 12 hours were selected for subsequent inflammatory reaction studies; The expression of LOC339524 protein was increased viadifferent concentrations of LPS treatment(

0.05) when compared with 0 ng/mL

and that increase was in a time-dependent manner at 100 ng/mL LPS concentration (

0.05). After 12 hours of LPS treatment

the expression of TNF-α and IL-6 mRNA and protein was significantly up-regulated in the siRNA group compared with the unrelated fragment group (

0.05). Conversely the expression of TNF-α and IL-6 mRNA and protein in the over-expression group was significantly lower than that in the normal group (

0.05).

Conclusion

LOC339524 is an inflammatory response factor that promotes the inflammatory response of bv2 cells to LPS.

关键词

Keywords

references

AUNGST S L, KABADI S V, THOMPSON S M, et al . Repeated mild traumatic brain injury causes chronic neuroinflammation, changes in hippocampal synaptic plasticity, and associated cognitive deficits[J ] . J Cereb Blood Flow Metab, 2014, 34(7): 1223-1232. DOI: 10.1038/jcbfm.2014.75.

WIMMER I, ZRZAVY T, LASSMANN H. Neuroinflammatory responses in experimental and human stroke lesions[J]. J Neuroimmunol, 2018, 323: 10-18. DOI: 10.1016/j.jneuroim.2018.07.003

[DE OLIVEIRA MANOEL A L, MACDONALD R L. Neuroinflammation as a target for intervention in subarachnoid hemorrhage[J ] . Front Neurol, 2018, 9: 292. DOI: 10.3389eur.2018.00292 http://dx.doi.org/10.3389eur.2018.00292 .

VEZZANI A, VIVIANI B. Neuromodulatory properties of inflammatory cytokines and their impact on neuronal excitability[J]. Neuropharmacology, 2015, 96(Pt A): 70-82. DOI: 10.1016/j.neuropharm.2014.10.027.

LI H, LI J, WANG Y, et al . Proteomic analysis of effluents from perfused human heart for transplantation: identification of potential biomarkers for ischemic heart damage[J ] . Proteome Sci, 2012, 10(1): 21. DOI: 10.1186/ 1477-5956- 10-21.

LONG Z H, LI H, CHEN F, et al . Cloning and sequence analysis of the LOC339524 gene in Sprague-Dawley rats[J ] . Genet Mol Res, 2015, 14(4): 16577-16584. DOI: 10.4238/2015.December.11.4.

FRIEND L D, SHAH D D, DEPPONG C, et al . Adose-dependent requirement for the proline motif of CD28 in cellular and humoral immunity revealed by a targeted knockin mutant[J ] . J Exp Med, 2006, 203(9): 2121-2133. DOI: 10.1084/jem.20052230.

龙宗泓, 李洪, 陈凤, 等.抗人LOC339524单克隆抗体的制备和应用[J].细胞与分子免疫学杂志, 2015, 31(12): 1693-1697, 1701.

LONG Z H, LI H, CHEN F, et al .Preparation and application of monoclonal antibody against human LOC339524 protein[J ] . Chin J Cell Mol Immunol, 2015, 31(12): 1693-1697, 1709.

KAUR C, SIVAKUMAR V, ZOU Z, et al . Microglia-derived proinflammatory cytokines tumor necrosis factor-alpha and interleukin-1beta induce Purkinje neuronal apoptosis via their receptors in hypoxic neonatal rat brain[J ] . Brain Struct Funct, 2014, 219(1): 151-170. DOI: 10.1007/s00429-012-0491-5..

NISHIOKU T, MATSUMOTO J, DOHGU S, et al . Tumor necrosis factor-alpha mediates the blood-brain barrier dysfunction induced by activated microglia in mouse brain microvascular endothelial cells[J ] . J Pharmacol Sci, 2010, 112(2): 251-254. DOI: 10.1254/jphs.09292sc.

FU Y Y, ZHANG F, ZHANG L, et al .Mangiferin regulates interleukin-6 and cystathionine-b-synthase in lipopolysaccharide-induced brain injury[J ] . Cell Mol Neurobiol, 2014, 34(5): 651-657. DOI: 10.1007/s10571-014-0039-8.

KWAN J, HORSFIELD G, BRYANT T, et al . IL-6 is a predictive biomarker for stroke associated infection and future mortality in the elderly after an ischemic stroke[J ] . Exp Gerontol, 2013, 48(9): 960-965. DOI: 10.1016/j.exger.2013.07.003.

白玉娟, 李琦军, 周红艳, 等.脂多糖干预时间与小胶质细胞生成IL-1β和TNF-α时效关系的研究[J].现代中西医结合杂志, 2016, 25(8): 828-831, 847. DOI: 10.3969/j.issn.1008-8849.2016.08.009.

BAI Y J, LI Q J, ZHOU H Y, et al . Time-effect relationship between lipopolysaccharide intervention time and IL-1β and TNF-α production by microglia[J ] . Mod J Integr Tradit Chin West Med, 2016, 25(8): 828-831, 847. DOI: 10.3969/j.issn.1008-8849.2016.0 8.009.

李乔俊, 王建文, 王卉, 等.脂多糖干预小胶质细胞炎症模型中血小板生成素与白细胞介素-6的表达及其相关性[J].实用儿科临床杂志, 2011, 26(4): 286-288. DOI: 10.3969/j.issn.1003-515X.2011.04.020.

LI Q J, WANG J W, WANG H, et al .Expression and correlation of thrombopoietin and interleukin-6 in lipopolysaccharide intervention model of microglia[J ] . Chin J Prac Pediatr, 2011, 26(4): 286-288. DOI: 10.3969/j.issn.1003-515X.2011.04.020.

DANIELS C C, COAN P, KING J, et al . The Proline-rich region of pneumococcal surface proteins A and C contains surface-accessible epitopes common to all pneumococci and elicits antibody-mediated protection against sepsis[J ] . Infect Immun, 2010, 78(5): 2163-2172. DOI: 10.1128/iai. 01199-09.

SHI J, ROSS C R, LETO T L, et al . PR-39, a proline-rich antibacterial peptide that inhibits phagocyte NADPH oxidase activity by binding to src homology 3 domains of p47 phox[J ] . Proc Natl Acad Sci USA, 1996, 93(12): 6014-6018. DOI: 10.1073/pnas.93.12. 6014.

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