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LOC339524基因介导脂多糖诱导小鼠小胶质bv2细胞系炎症因子的表达
LOC339524 gene mediates lipopolysaccharide-induced expression of inflammatory cytokines in mouse microglia bv2 cell line
- 第三军医大学学报 2019年41卷第4期 页码：289-295
- 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：
[ 李洪，E-mail：lh78553@163.com LI Hong, E-mail:lh78553@163.com
- 基金信息：国家自然科学基金面上项目(81571870)
DOI：10.16016/j.1000-5404.201810166
中图分类号： R392-33;R392.11;R392.13收稿：2018-10-25，
修回：2018-12-07，
纸质出版：2019-02-28
- 稿件说明：
移动端阅览
唐相龙, 龙宗泓, 段光友, 等. LOC339524基因介导脂多糖诱导小鼠小胶质bv2细胞系炎症因子的表达[J]. 第三军医大学学报, 2019,41(4):289-295.
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.
唐相龙, 龙宗泓, 段光友, 等. LOC339524基因介导脂多糖诱导小鼠小胶质bv2细胞系炎症因子的表达[J]. 第三军医大学学报, 2019,41(4):289-295. DOI： 10.16016/j.1000-5404.201810166.
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
2
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
2
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
2
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(
P
<
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(
P
<
0.05)，100 ng/mL LPS处理后呈时间依赖性增加(
P
<
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(
P
<
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(
P
<
0.05)。
结论
2
LOC339524基因介导了脂多糖诱导bv2细胞的炎症反应。
Abstract
Objective
2
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
2
Mouse microglia bv2 cells were treated with 100 ng/mL LPS for 3
6
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
6
9
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
2
Compared with blank control
LPS treatment increased TNF-α
IL-6 mRNA and protein expression at different times (
P
<
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(
P
<
0.05) when compared with 0 ng/mL
and that increase was in a time-dependent manner at 100 ng/mL LPS concentration (
P
<
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 (
P
<
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 (
P
<
0.05).
Conclusion
2
LOC339524 is an inflammatory response factor that promotes the inflammatory response of bv2 cells to LPS.
关键词
Keywords
references
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