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:
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.
Effect of long non-coding RNA Rpph1 on expression of inflammatory cytokines in mesangial cells under high glucose condition
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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