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长链非编码RNA Gm4419通过NF-κB信号通路调控高糖下系膜细胞炎症因子的表达
LncRNA Gm4419 regulates inflammatory cytokines expression
via NF-κB signaling pathway in mesangial cells under high glucose condition- 第三军医大学学报 2018年40卷第7期 页码：577-583
- 作者机构：
1.400016 重庆，重庆医科大学基础医学院：细胞生物学及遗传学教研室
2.400016 重庆，重庆医科大学基础医学院：生物信息学教研室
3.400016 重庆，重庆医科大学基础医学院：实验教学中心
- 作者简介：
[张政，E-mail：zhangzheng92@163.com ZHANG Zheng, E-mail: zhangzheng92@163.com
- 基金信息：国家自然科学基金面上项目(81570747)
DOI：10.16016/j.1000-5404.201711099
中图分类号： R322.61;R329.28;R394.2收稿：2017-11-13，
修回：2017-12-30，
纸质出版：2018-04-15
- 稿件说明：
移动端阅览
姜文豪, 易红, 彭睿, 等. 长链非编码RNA Gm4419通过NF-κB信号通路调控高糖下系膜细胞炎症因子的表达[J]. 第三军医大学学报, 2018,40(7):577-583.
Wenhao JIANG, Hong YI, Rui PENG, et al. LncRNA Gm4419 regulates inflammatory cytokines expression
via NF-κB signaling pathway in mesangial cells under high glucose condition[J]. Journal of Third Military Medical University, 2018, 40(7): 577-583.姜文豪, 易红, 彭睿, 等. 长链非编码RNA Gm4419通过NF-κB信号通路调控高糖下系膜细胞炎症因子的表达[J]. 第三军医大学学报, 2018,40(7):577-583. DOI： 10.16016/j.1000-5404.201711099.
Wenhao JIANG, Hong YI, Rui PENG, et al. LncRNA Gm4419 regulates inflammatory cytokines expression
via NF-κB signaling pathway in mesangial cells under high glucose condition[J]. Journal of Third Military Medical University, 2018, 40(7): 577-583. DOI： 10.16016/j.1000-5404.201711099.
摘要
目的
2
探讨长链非编码RNA Gm4419通过核转录因子-kappa B (nuclear factor-kappa B
NF-κB)信号通路对高糖培养的肾小球系膜细胞炎症因子表达的影响。
方法
2
在高、低糖培养的小鼠肾小球系膜细胞中，采用RT-qPCR检测Gm4419的亚细胞分布及NF-κB亚基p50和p65的表达水平；采用RT-qPCR及免疫荧光检测上、下调Gm4419后炎症相关因子TNF-α和MCP-1的表达；采用生物信息技术分析Gm4419与p50和p65关系；采用Western blot检测上、下调Gm4419后细胞核中p50和p65的表达水平，以及检测特异性p50抑制剂对过表达Gm4419影响MCP-1表达的作用。
结果
2
Gm4419在肾小球系膜细胞高糖组表达水平显著高于低糖组(
P
<
0.05)，且主要分布在
系膜细胞的细胞质中。同时，在低糖组过表达Gm4419后MCP-1、TNF-α表达水平显著增加(
P
<
0.01)；而在高糖组沉默Gm4419后MCP-1、TNF-α表达水平显著减少(
P
<
0.01)。此外，Gm4419与NF-κB亚基p50和p65均存在潜在结合位点，过表达Gm4419后胞核p50和p65蛋白表达水平显著增加，NF-κB活化增强(
P
<
0.01)；而沉默Gm4419后胞核p50和p65蛋白表达水平显著降低，NF-κB活化减弱(
P
<
0.01)，且过表达Gm4419增加MCP-1表达的效应可被特异性p50抑制剂阻断。
结论
2
Gm4419可能在促进高糖下系膜细胞炎症因子表达中扮演重要角色，其机制可能涉及NF-κB信号通路。
Abstract
Objective
2
To investigate the mechanism of long non-coding RNA (lncRNA) Gm4419 regulating the expression of inflammatory cytokines in glomerular mesangial cells under high glucose condition
via
mediating nuclear factor-kappa B (NF-κB) signaling pathway.
Methods
2
The subcellular distribution of Gm4419 and expression of NF-κB subunits p50 and p65 were detected by RT-qPCR in mouse glomerular mesangial cells (MCs) under high or low glucose condition. Moreover
RT-qPCR and immunofluorescence assay were used to measure the expression of MCP-1 and TNF-α when Gm4419 was up-or down-regulated. Additionally
bioinformatics analysis was carried out to predict the relationship between Gm4419 and NF-κB. Western blot assay was used to determine the expression of p50 and p65 in the nucleus of MCs
and the effect of specific p50 inhibitor on the expression of MCP-1 was studied in MCs with Gm4419 over-expression.
Results
2
The expression of Gm4419 was significantly higher in MCs under high glucose condition than those under low condition (
P
<
0.05)
and Gm4419 mainly located in the cytoplasm. The expression levels of MCP-1 and TNF-α were significantly increased in the low glucose group when Gm4419 was over-expressed (
P
<
0.01)
while the levels were decreased in high glucose group when Gm4419 was silenced (
P
<
0.01). Moreover
the binding sites of Gm4419 and p
50 and p65 were predicted
and over-expression of Gm4419 resulted in the increases in p50 and p65 expression levels and enhancement in NF-κB activity (
P
<
0.01). However
silencing Gm4419 induced opposite effects (
P
<
0.01). Additionally
specific p50 inhibitor could block the up-reguation of MCP-1 induced by Gm4419 over-expression.
Conclusion
2
Gm4419 may play an important role in the enhanced expression of inflammatory cytokines in mesangial cells under high glucose condition
and its mechanism may be related to the NF-κB signaling pathway.
关键词
Keywords
references
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