Protective effect of miR-146 against kidney injury in diabetic nephropathy rats through mediating the NF-κB signaling pathway
Authors: X Ci
L-F Meng
L-H Liu
H-Y Yu
X-H Lu
Z Zhuo
Journal: European review for medical and pharmacological sciences (2020)
Discipline: PHARMACOLOGY & TOXICOLOGY
Keywords: miR-146; kidney injury; diabetic nephropathy; NF-κB signaling pathway; rats
Affiliations: Department of Nephrology, The Second Hospital of Jilin University, Changchun, China. .

Abstract:  Objective: To study the protective effect of micro ribonucleic acid (miR)-146 against kidney injury in diabetic nephropathy (DN) rats through the nuclear factor-κB (NF-κB) signaling pathway. Materials and methods: In this experiment, 30 adult Sprague-Dawley rats with 5-6 weeks old and weighing 20-30 g were selected and randomly divided into control group (n=10), model group (n=10), and miR-146 Mimic group (n=10, DN rat model + miR-146 Mimic). The serum levels of creatinine (Cr) and blood urea nitrogen (BUN) in the three groups were detected using the full-automatic biochemical analyzer. The protein expression levels of phosphorylated-inhibitor of NF-κB (p-IκB), p-P65, P65, and Tubulin were detected via Western blotting. The messenger RNA (mRNA) of P65 was determined using quantitative Polymerase Chain Reaction (qPCR). Positive expression of p-IκB in tissues was determined using immunohistochemistry. Moreover, the contents of inflammatory factors tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-6 were detected using the enzyme-linked immunosorbent assay (ELISA) kits. Finally, the apoptosis was detected through Annexin V-fluorescein isothiocyanate (FITC) and propidium iodide (PI) dual-fluorescence labeling. Results: The serum levels of Cr and BUN were significantly higher in the model group than those in the control group (p&lt;0.01), while they were significantly lower in miR-146 Mimic group than those in the model group (p&lt;0.05). The levels of p-IκB and p-P65/P65 significantly increased in the model group compared with those in the control group (p&lt;0.01), while they remarkably declined in the miR-146 Mimic group compared with those in the model group (p&lt;0.05). The results of qPCR showed that the mRNA level of P65 had no significant difference among the three groups (p&gt;0.05). The immunohistochemical assay showed that the positive expression of p-IκB in tissues was consistent with those of the protein level as Western blotting revealed. The rats in the model group had evidently increased levels of TNF-α, IL-1β, and IL-6 compared with the control group (p&lt;0.01), while miR-146 Mimic group had evidently decreased levels of them compared with the model group (p&lt;0.01). Finally, apoptosis was enhanced in the model group compared with that in the control group, while it was remarkably inhibited in the miR-146 Mimic group. Conclusions: MiR-146 can inhibit the NF-κB signaling pathway, lower the levels of TNF-α, IL-1β, and IL-6, and reduce the apoptosis, thereby exerting a protective effect against kidney injury in DN. 

Retracted: Yes

PubPeer comments:
 Figure 4.  Effect of miR-146 on apoptosis of kidney tissues. A, Apoptosis determined using Annexin V-FITC and PI dualfluorescence labeling and flow cytometry . ![file] Figure 5 from [Jiang et al (2020)] .  Effect of miR-210 on apoptosis level in each group. A, Apoptosis detected via Annexin V-FITC and PI double labeling . ![file] , 
  Control and  Model panels of Fig 4 re-appear as Fig 5A of [Han et al (2020)] ,  Impact of lncRNA MEG3 on cell apoptosis. A, Impact of lncRNA MEG3 on the apoptosis of RSC-364 cells detected via FITC PI double staining and flow cytometer. ![] , 
 #1 Fig 4 again - now compared with (below) three panels from Fig 11 of [Zhang et al (2018)] ,  Apoptosis rate of cardiomyocytes in each group after transfection. Notes: A, apoptotic scatter plot . ![file] Similarities are greatest in the left-hand quadrants of each pair. , 
 Retracted [November 2021] . -------------------------  The article u201cProtective effect of miR-146 against kidney injury in diabetic nephropathy rats through mediating the NF-u03baB signaling pathwayu201d, H.-Y. Yu, L.-F. Meng, X.-H. Lu, L.-H. Liu, X. Ci, Z. Zhuo, published in Eur Rev Med Pharmacol Sci 2020; 24 (6): 3215-3222u2013DOI: 10.26355 eurrev_202003_20688u2013PMID: 32271439, has been retracted by the authors due to some inaccuracies (some data cannot be repeated by further research). The Publisher apologizes for any inconvenience this may cause. ,