Effect of over-expressed miR-148a on secretion and expression of cytokines in human adipose-derived mesenchymal stem cell differentiation process

Abstract

Objective To investigate the effect of over-expressed miR-148a on secretion and expression of inflammatory and insulin sensitive protein, such as interleukin 6 (IL-6), leptin, resistin and tumor necrosis factor α (TNF-α), during human adipose-derived mesenchymal stem cells (hMSCs-Ad) differentiation. Methods hMSCs-Ad was infected with the lentivirus of hsa-miR-148a, then the secretion of inflammatory cytokines (IL-6, TNF-α) and insulin sensitive protein (leptin, resistin) during differentiation process.The cells were collected at 0, 4, 10 and 15 days for RNA extration.The expression levels of IL-6, leptin, resistin and TNF-α were measured by using enzyme-linked immunosorbent assay, IL-6 and leptin mRNA were examined by using real-time PCR. Results Compared with the control group, after hMSCs-Ad were infected with over-expressed miR-148a lentivirus, both secretion of IL-6 [day10: (50.92±14.93) ng/L vs (136.86±35.53) ng/L, t=3.86, P=0.018; day 15: (123.64±18.76) ng/L vs (178.59±7.54) ng/L, t=1.56, P=0.029]and leptin day 4: (12.28±0.56) ng/L vs (14.38±0.84) ng/L, t=3.06, P=0.038; day 15: (34.45±4.30) ng/L vs (115.85±28.81) ng/L, t=4, 79, P=0.009] were reduced, while the concentration of resistin was higher than that of the control group and the concentration of resistin was less 0.20 μg/L in both groups, and TNF-α did not express, in hMSCs-Ad differentiation.Similarly, IL-6(day 4: 0.041±0.018 vs 0.089±0.002, t=4.68, P=0.009; day 15: 0.138±0.015 vs 0.284±0.069, t=3.55, P=0.024) and leptin (day 10: 0.163±0.039 vs 0.497±0.042, t=10.13, P=0.001; day 15: 0.259±0.033 vs 1.146±0.346, t=4.41, P=0.012) mRNA expression levels were reduced in overexpressed miR-148a group. Conclusions It suggests that miR-148a can improve hMSCs-Ad inflammation status and improve insulin resistance through regulating the IL-6, leptin, resistin and TNF-α expressions, which indicates that miR-148a play a role in the process of obesity related inflammation and insulin sensitivity. Key words: miR-148a; Human adipose-derived mesenchymal stem cell; Adipokines; Inflammation; Obesity