浏览全部资源
扫码关注微信
长链非编码RNA RP11-288L9.1在系统性红斑狼疮中的作用及其机制研究
Role of long non-coding RNA RP11-288L9.1 in systemic lupus erythematosus and underlying mechanism
- 第三军医大学学报 2021年43卷第13期 页码：1204-1211
- 作者机构：
400038 陆军军医大学(第三军医大学)第一附属医院皮肤科
- 作者简介：
[游弋，E-mail: youyi_cq@163.comYOU Yi, E-mail: youyi_cq@163.com
- 基金信息：国家自然科学基金面上项目(81673058);重庆市基础科学与前沿技术研究一般项目(cstc2017jcyjAX0251)
DOI：10.16016/j.1000-5404.202101010
中图分类号： R392.13;R392.32;R593.24收稿：2021-01-03，
修回：2021-02-05，
纸质出版：2021-07-15
- 稿件说明：
移动端阅览
赵承磊, 赵兴旺, 郭俊恺, 等. 长链非编码RNA RP11-288L9.1在系统性红斑狼疮中的作用及其机制研究[J]. 第三军医大学学报, 2021,43(13):1204-1211.
Chenglei ZHAO, Xingwang ZHAO, Junkai GUO, et al. Role of long non-coding RNA RP11-288L9.1 in systemic lupus erythematosus and underlying mechanism[J]. Journal of Third Military Medical University, 2021, 43(13): 1204-1211.
赵承磊, 赵兴旺, 郭俊恺, 等. 长链非编码RNA RP11-288L9.1在系统性红斑狼疮中的作用及其机制研究[J]. 第三军医大学学报, 2021,43(13):1204-1211. DOI： 10.16016/j.1000-5404.202101010.
Chenglei ZHAO, Xingwang ZHAO, Junkai GUO, et al. Role of long non-coding RNA RP11-288L9.1 in systemic lupus erythematosus and underlying mechanism[J]. Journal of Third Military Medical University, 2021, 43(13): 1204-1211. DOI： 10.16016/j.1000-5404.202101010.
摘要
目的
2
探讨长链非编码RNA(long non-conding RNA，lncRNA)RP11-288L9.1在系统性红斑狼疮(systemic lupus erythematosus，SLE)中的作用及机制。
方法
2
通过转录组测序方法(RNA-seq)筛选获得SLE患者外周血单核细胞(peripheral blood mononuclear cell
PBMCs)中差异表达的lncRNAs，采用实时定量PCR(quantitative real-time PCR，qRT-PCR)技术验证8例SLE患者和8例健康对照PBMCs中差异表达的lncRNA的表达水平。运用荧光原位杂交FISH技术检测RP11-288L9.1的细胞定位；设计并构建RP11-288L9.1敲低和过表达重组慢病毒，转染构建RP11-288L9.1敲低和过表达模型，采用qRT-PCR验证其转染效率。分别采用CCK-8和Annexin V-FITC细胞凋亡检测试剂盒检测慢病毒转染后巨噬细胞的增殖和凋亡能力，qRT-PCR检测敲低和高表达RP11-288L9.1后，巨噬细胞相关炎症细胞因子IL-1β、IL-6、IL-4、IL-10、TGF-β1的表达。
结果
2
通过转录组数据发现6个差异表达的lncRNA(
P
<
0.05)。qRT-PCR验证RP11-288L9.1表达显著上调(
P
<
0.01)。RP11-288L9.1定位在巨噬细胞胞浆中。过表达RP11-288L9.1可促进巨噬细胞的凋亡并抑制增殖(
P
<
0.05)，相关炎症因子IL-1β、IL-6表达显著上升，IL-4、IL-10、TGF-β1表达显著下降(
P
<
0.001)；敲低RP11-288L9.1可促进巨噬细胞的增殖并抑制凋亡(
P
<
0.05)，相关炎症因子IL-1β、IL-6表达显著下降(
P
<
0.01)，IL-4、IL-10、TGF-β1表达显著上升(
P
<
0.01)。
结论
2
RP11-288L9.1在SLE患者PBMCs中高表达，并在巨噬细胞中通过影响炎症细胞因子的合成来调控免疫反应，这可能是SLE发病机制之一。
Abstract
Objective
2
To investigate the role of long non-coding RNA (lncRNA) RP11-288L9.1 in systemic lupus erythematosus (SLE) and its potential mechanism.
Methods
2
Transcriptome sequencing (RNA-seq) was used to screen the differentially expressed lncRNAs in peripheral blood mononuclear cells (PBMCs) of SLE patients
and the expression levels of these lncRNAs in 8 pairs of SLE patients and healthy controls were further verified by qRT-PCR. The knockdown as well as over-expression models of RP11-288L9.1 were constructed respectively by transfecting recombinant lentivirus into macrophages
and the transfection efficiency was determined subsequently. The subcellular distribution of RP11-288L9.1 was detected by fluorescence
in situ
hybridization (FISH) technique. Moreover
CCK-8 assay and Annexin V-FITC apoptosis detection kit were applied to examine the proliferation and apoptosis of macrophages in both models
and qRT-PCR was conducted to detect the expression of macrophage-related inflammatory cytokines
IL-1β
IL-6
IL-4
IL-10 and TGF-β1 after the transfection.
Results
2
Six differentially expressed lncRNAs were found (
P
<
0.05)
in which RP11-288L9.1 was significantly up-regulated (
P
<
0.01)
and mainly located in the cytoplasm of macrophages. The over-expression of RP11-288L9.1 promoted the apoptosis and inhibited the proliferation of macrophages (
P
<
0.05)
increasing the levels of IL-1β and IL-6
and decreasing those of IL-4
IL-10 and TGF-β1 (
P
<
0.01). In contrast
the knockdown of RP11-288L9.1 significantly induced the proliferation of macrophages and inhibited the apoptosis (
P
<
0.05)
with lowered levels of IL-1β and IL-6 and elevated IL-4
IL-10 and TGF-β1 (
P
<
0.01).
Conclusion
2
RP11-288L9.1 is highly expressed in the PBMCs of SLE patients
and regulates immune response in macrophages by affecting the synthesis of inflammatory cytokines
which may contribute to the occurrence of SLE.
关键词
Keywords
references
KIRIAKIDOU M, CHING C L. Systemic lupus erythematosus[J]. Ann Intern Med, 2020, 172(11): ITC81-ITC96. DOI: 10.7326/AITC202006020.
DURCAN L, O'DWYER T, PETRI M. Management strategies and future directions for systemic lupuserythematosus in adults[J]. Lancet, 2019, 393(10188): 2332-2343. DOI:10.1016/s0140-6736(19)30237-5.
YAO R W, WANG Y, CHEN L L. Cellular functions of long noncoding RNAs[J]. Nat Cell Biol, 2019, 21(5): 542-551. DOI:10.1038/s41556-019-0311-8.
SUN Q, HAO Q, PRASANTH K V. Nuclear long noncoding RNAs: key regulators of gene expression[J]. Trends Genet, 2018, 34(2): 142-157. DOI:10.1016/j.tig.2017.11.005.
VERGADI E, VAPORIDI K, TSATSANIS C. Regulation of endotoxin tolerance and compensatory anti-inflammatory response syndrome by non-coding RNAs[J]. Front Immunol, 2018, 9: 2705. DOI:10.3389/fimmu.2018.02705.
WU Y, ZHANG F, MA J, et al . Association of large intergenic noncoding RNA expression with disease activity and organ damage in systemic lupus erythematosus[J ] . Arthr Res Ther, 2015, 17(1): 131. DOI: 10.1186/s13075-015-0632-3.
WU G C, PAN H F, LENG R X, et al . Emerging role of long noncoding RNAs in autoimmune diseases[J ] . Autoimmun Rev, 2015, 14(9): 798-805. DOI:10.1016/j.autrev.2015.05.004.
LONG H, YIN H, WANG L, et al . The critical role of epigenetics in systemic lupus erythematosus and autoimmunity[J ] . J Autoimmun, 2016, 74: 118-138. DOI:10.1016/j.jaut.2016.06.020.
LI L J, ZHAO W, TAO S S, et al . Comprehensive long non-coding RNA expression profiling reveals their potential roles in systemic lupus erythematosus[J ] . Cell Immunol, 2017, 319: 17-27. DOI:10.1016/j.cellimm.2017.06.004.
CARPENTER S, AIELLO D, ATIANAND M K, et al . A long noncoding RNA mediates both activation and repression of immune response genes[J ] . Science, 2013, 341(6147): 789-792. DOI:10.1126/science.1240925.
CASTELLANOS-RUBIO A, FERNANDEZ-JIMENEZ N, KRATCHMAROV R, et al . A long noncoding RNA associated with susceptibility to celiac disease[J ] . Science, 2016, 352(6281): 91-95. DOI:10.1126/science.aad0467.
CHAN J, ATIANAND M, JIANG Z, et al . Cutting edge: a natural antisense transcript, AS-IL1α, controls inducible transcription of the proinflammatory cytokine IL-1α[J ] . J Immunol, 2015, 195(4): 1359-1363. DOI:10.4049/jimmunol.1500264.
HUME D A. The many alternative faces of macrophage activation[J]. Front Immunol, 2015, 6: 370. DOI:10.3389/fimmu.2015.00370.
CHEUNSUCHON B, INCHAROEN P, CHAWANASUN-TORAPOJ R, et al . Glomerular macrophage is an indicator of early treatment response in diffuse proliferative lupus nephritis[J ] . J Med Assoc Thai, 2013, 96(Suppl 2): S246-S251.
CHALMERS S A, CHITU V, HERLITZ L C, et al . Macrophage depletion ameliorates nephritis induced by pathogenic antibodies[J ] . J Autoimmun, 2015, 57: 42-52. DOI:10.1016/j.jaut.2014.11.007.
YOON J, UM H N, JANG J, et al . Eosinophil activation by Toll-like receptor 4 ligands regulates macrophage polarization[J ] . Front Cell Dev Biol, 2019, 7: 329. DOI:10.3389/fcell.2019.00329.
CASTELLANOS-RUBIO A, KRATCHMAROV R, SEBASTIAN M, et al . Cytoplasmic form of carlr lncRNA facilitates inflammatory gene expression upon NF-κB activation[J ] . J Immunol, 2017, 199(2): 581-588. DOI:10.4049/jimmunol.1700023.
CHEN X, DU J, ZHU Y, et al . Comprehensive analysis of lncRNA and mRNA expression profiles in macrophages activated by Actinidia eriantha polysaccharide[J ] . Int J Biol Macromol, 2019, 136: 980-993. DOI:10.1016/j.ijbiomac.2019.06.091.
LI H, JIANG T, LI M Q, et al . Transcriptional regulation of m acrophages polarization by microRNAs[J ] . Front Immunol, 2018, 9: 1175. DOI:10.3389/fimmu.2018.01175.
YIN L, DAI Q, JIANG P, et al . Manganese exposure facilitates microglial JAK2-STAT3 signaling and consequent secretion of TNF-α and IL-1β to promote neuronal death[J ] . Neurotoxicology, 2018, 64: 195-203. DOI: 10.1016/j.neuro.2017.04.001.
徐雅楠, 于嘉霖, 马臣杰, 等. Lnc RNA-GAS5对BCG感染小鼠巨噬细胞后细胞坏死的调控作用[J]. 微生物学通报, 2019, 46(4): 858-868. DOI: 10.13344/j.microbiol.china.180317.
XU Y N, YU J L, MA C J, et al . Regulation of LncRNA-GAS5 on necrosis of macrophage induced by Bacillus Calmette-Guérin[J ] . Microbio China, 2019, 46(4): 858-868. DOI: 10.13344/j.microbiol.china.180317.
0
浏览量
0
下载量
0
CSCD
- 网络PDF下载
- Meta-XML下载
- BibTeX下载
- Endnote下载
- RefMan 下载
- NoteExpress下载
- NoteFirst下载
关联资源
相关文章
相关作者
相关机构
- 技术支持由北京北大方正电子有限公司提供 京ICP备09064830号-19
京公网安备11010802024621 - 本系统建议在Chrome、 IE9+ 以上版本浏览器阅读本站内容，360浏览器请切换至极速模式
- Cookies帮助我们提供服务并提供个性化体验。使用本网站，即表示您同意我们使用Cookies
