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长链非编码RNA Rpph1对高糖培养的肾小球系膜细胞炎症因子表达的影响
Effect of long non-coding RNA Rpph1 on expression of inflammatory cytokines in mesangial cells under high glucose condition
- 第三军医大学学报 2019年41卷第1期 页码：33-40
- 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：分子医学与肿瘤研究中心
- 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.1000-5404.201808070
中图分类号： R394.3;R587.2;R692.02收稿：2018-08-09，
修回：2018-10-04，
纸质出版：2019-01-15
- 稿件说明：
移动端阅览
张攀扬, 孙艳, 彭睿, 等. 长链非编码RNA Rpph1对高糖培养的肾小球系膜细胞炎症因子表达的影响[J]. 第三军医大学学报, 2019,41(1):33-40.
Panyang ZHANG, Yan SUN, Rui PENG, et al. Effect of long non-coding RNA Rpph1 on expression of inflammatory cytokines in mesangial cells under high glucose condition[J]. Journal of Third Military Medical University, 2019, 41(1): 33-40.
张攀扬, 孙艳, 彭睿, 等. 长链非编码RNA Rpph1对高糖培养的肾小球系膜细胞炎症因子表达的影响[J]. 第三军医大学学报, 2019,41(1):33-40. DOI： 10.16016/j.1000-5404.201808070.
Panyang ZHANG, Yan SUN, Rui PENG, et al. Effect of long non-coding RNA Rpph1 on expression of inflammatory cytokines in mesangial cells under high glucose condition[J]. Journal of Third Military Medical University, 2019, 41(1): 33-40. DOI： 10.16016/j.1000-5404.201808070.
摘要
目的
2
探讨长链非编码RNA (long non-coding RNA，lncRNA) Rpph1对糖尿病肾病(diabetic nephropathy，DN)肾小球系膜细胞炎症因子的影响。
方法
2
采用荧光原位杂交(fluorescence in situ hybridization
FISH)检测Rpph1细胞定位，实时定量PCR (quantitative real-time PCR，qRT-PCR)检测Rpph1在DN小鼠肾脏组织和系膜细胞中的水平；生物信息学技术预测Rpph1的编码蛋白能力。构建Rpph1的过表达质粒，并设计合成Rpph1的siRNAs，用qRT-PCR验证其转染效率。在高糖培养的肾小球系膜细胞中转染Rpph1-siRNA；在低糖培养的肾小球系膜细胞中转染Rpph1过表达质粒，通过Western blot检测转染Rpph1的siRNA和过表达Rpph1后肿瘤坏死因子(tumor necrosis factor α，TNF-α)以及单核细胞趋化蛋白-1(macrophage cationic peptide 1，MCP-1)的表达情况。
结果
2
与正常小鼠比较，Rpph1在DN小鼠肾脏组织中显著上调(
P
<
0.01)；同样，高糖培养的系膜细胞中Rpph1表达较低糖培养的系膜细胞显著上调(
P
<
0.01)，其主要定位于系膜细胞的细胞质。此外，生物信息学证实Rpph1不具有编码蛋白的能力。且过表达Rpph1后炎
症相关因子TNF-α和MCP-1的水平明显上调(
P
<
0.05)；在下调Rpph1后，炎症相关因子TNF-α和MCP-1的表达也随之下降(
P
<
0.05)。
结论
2
在DN小鼠肾脏组织和高糖环境下的系膜细胞中lncRNA Rpph1显著高表达，且增强炎症相关因子的表达。Rpph1可能与糖尿病肾病炎症的发生相关。
Abstract
Objective
2
To investigate the effect of long non-coding RNA (lncRNA) Rpph1 on the expression of inflammatory cytokines in mouse glomerular mesangial cells (MCs) from mouse model of diabetic nephropathy (DN).
Methods
2
The subcellular distribution and expression of Rpph1 in the renal tissue of DN mice and the isolated glomerular mesangial cells (MCs) were detected by fluorescence
in situ
hybridization (FISH) and quantitative real-time PCR (qRT-PCR). The coding potential of Rpph1 was evaluated by using ORF Finder and Coding Potential Calculator 2. The overexpressed plasmid and siRNA sequence of Rpph1 were constructed and designed and synthesized
and then transfected into MCs under low and high glucose conditions
respectively
the transfection effect of pcDNA3.1 (+)-Rpph1 and Rpph1 siRNAs was tested by qRT-PCR. Moreover
Western blotting was used to detect the expression of inflammatory factors TNF-α and macrophage cationic peptide 1 (MCP-1).
Results
2
The expression level of Rpph1 was significantly higher in the renal tissue of DN mice than that of healthy control (
P
<
0.01). The molecule was mainly located in the cytoplasm of MCs
and its level was as well increased in MCs under high glucose condition than those under low glucose (
P
<
0.01). Moreover
bioinformatics data identified that Rpph1 could not code proteins. Additionally
Rpph1 overexpression up-regulated significantly the levels of TNF-α and MCP-1 (
P
<
0.05)
while its knockdown induced the decreases in TNF-α and MCP-1 levels (
P
<
0.05).
Conclusion
2
LncRNA-Rpph1 is highly expressed in the renal tissues of DN mice and in MCs cultured in high glucose
and its higher level enhances the expression of inflammatory cytokines. Rpph1 may be associated with the pathogenesis of inflammation in DN.
关键词
Keywords
references
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