MicroRNA (miR)-590-3p alleviates high-glucose induced renal tubular epithelial cell damage by targeting C-X3-C motif chemokine ligand 1 (CX3CL1) in diabetic nephropathy

Figures & data

Table 1. Comparison of the baseline data of subject

Parameters	Health (n = 42)	T2DM (n = 50)	DN (n = 47)
Age (year)	47.40 ± 16.18	48.12 ± 16.51	54.11 ± 16.44
Gender (female/male)	22/20	20/30	23/24
BMI (kg/m^2)	27.14 ± 1.76	27.16 ± 2.54	27.64 ± 2.06
SBP (mmHg)	106.32 ± 8.06	122.44 ± 12.56*	135.19 ± 12.18* ^#
DBP (mmHg)	73.68 ± 5.89	77.22 ± 8.89*	84.72 ± 9.90* ^#
TC (mmol/L)	179.54 ± 25.92	178.14 ± 38.61	191.32 ± 41.32
LDL cheolesterol (mg/dL)	108.79 ± 20.89	115.09 ± 35.04	113.67 ± 36.88
HDL cheolesterol (mg/dL)	64.62 ± 13.05	56.22 ± 15.55*	50.76 ± 11.11* ^#
Triglyceride (mmol/L)	169.82 ± 11.48	170.46 ± 18.25	168.98 ± 19.66
FBG (mg/dL)	95.13 ± 10.29	212.10 ± 99.22*	193.90 ± 105.93*
hs-CRP (mg/L)	0.49 ± 0.11	9.83 ± 3.82*	7.94 ± 4.32* ^#
HbA1c (%)	-	9.01 ± 1.93*	10.30 ± 2.80* ^#
eGFR	100.42 ± 10.94	103.74 ± 12.29	56.19 ± 16.04* ^#
Albuminuria (mg/24 h)	-	7.94 ± 3.00*	130.58 ± 55.38* ^#

Note: BMI, body mass index; SBP, systolic blood pressure; DBP, Diastolic blood pressure; TC, total cholesterol; LDL, Low-density lipoprotein; HDL, high density lipoprotein; FBG, fasting blood glucose; hs-CRP, high-sensitivity C-reactive protein; eGFR, estimated glomerular filtration rate; HbA1c, glycated hemoglobin; T2DM, Type 2 diabetes mellitus; DN, diabetic nephropathy * P < 0.05, compared with Healthy control; ^# P < 0.05, compared with T2DM patients.

Figure 1. Expression and clinical correlation analysis of miR-590-3p in participants. (a) Compared with healthy controls and T2DM patients, miR-590-3p is marked to be reduced in DN patients. Spearman correlation coefficient analysis of serum miR-590-3p and DN clinical index (b) Albuminuria (c) eGFR. note: compared with healthy, *** means P < 0.001; Compared with T2DM, ### means P < 0.001.

Figure 2. To evaluate the diagnostic ability of serum miR-590-3p for DN. (a) miR-590-3p can significantly identify DN patients from healthy individuals. The AUC of ROC was 0.958, the sensitivity was 92.90% and the specificity was 87.23%. (b) miR-590-3p could distinguish DN patients from T2DM patients, and the AUC of ROC was 0.859, the sensitivity and the specificity were 92.0% and 82.98%, respectively.

Figure 3. Effects of miR-590-3p level on HG-induced HK-2 proliferation, apoptosis and ROS damage. (a) mRNA levels of miR-590-3p at HG concentration gradient (0, 10, 20, 30, 40 mM). (b) miR-590-3p mRNA levels at 30 mM HG-induced under time gradient (0, 12, 24, 48 h). (c) The miR-590-3p mRNA levels of transfected with miR-590-3p mimic under HG-induced in the HK-2 cells. The elevated miR-590-3p regulates HG-induced cell proliferation (d), apoptosis (e), and oxidative stress (f). Note: Compared with 0 mM or 0 h or control, * means P < 0.05, ** means P < 0.01, *** means P < 0.001; Compared with HG + mimic NC, ## means P < 0.01; ### means P < 0.001.

Figure 4. miR-590-3p level affected HG-induced inflammation in HK-2 cells. (a) IL-6, (b) IL-18, and (c) TNF-α were increased in the HG-induced group, and (d) IL-10 was lower by the HG-induced. However, elevated miR-590-3p signature alleviates the levels of these inflammatory factors. Note: Compared with NG group, *** means P < 0.001; Compared with HG + mimic NC group, ### means P < 0.001.

Figure 5. CX3CL1 was the target of miR-590-3p in DN. (a) Predicted binding sites analyzed by online software. (b) The consequence miR-590-3p mimic on luciferase activity in the recombinant plasmid was examined by luciferase assay. (c) Serum CX3CL1 mRNA levels in DN patients. (d) Correlation analysis of miR-590-3p and CX3CL1 in patients’ serum. (e) miR-590-3p can regulate the mRNA levels of CX3CL1 in HK-2. Note: compared with mimic NC or healthy, *** means P < 0.001; compared with inhibitor NC, ### means P < 0.001.