The Role Of MicroRNA-214 In Ischemia-Reperfusion Injury Of Human Renal Tubular Epithelial Cells

objective To study the role of micro RNA- 214(mi R- 214) in ischemia-reperfusion injury of human renal tubular epithelial cells and the possible mechanism. Methods Human renal tubular epithelial cells(HK- 2) were used to establish ischemia-reperfusion injury as an in vitro cell model, the cells were devided into Sham Group and IRI Group(IRI 0h?8h?12h?24h?48h);then HK- 2 cell transfected with mi R- 214 mimics(Group mimic), the mi R- 214 inhibitor(Group inhibitor) and meaningless mi R(Group mimic NC and Group inhibitor NC),and one group with non-transfeced(Group IRI); then start an ischemia reperfusion(12h), to analysis HK-2 cells with mi R- 214 m RNA, Claudin-2 protein expression level and cell viability,cell apoptosis rate of each group. Results(1)During the ischemia-reperfusion injury in HK- 2 cells, the mi R- 214 expression quantity of IRI 12 h were the highest, m RNA ? Claudin-2 protein expression quantity were minimum, and cell vitality were the lowest, cell apoptosis rate were also the highest;(2) Mi R-214 was significantly increased in mimic group, for 1402 times of the baseline values. The mi R- 214 increased level of IRI group, inhibitor group, mimic- NC group, inhibitor- NC group were 10.2,9.8, 10.6,10.4 times of baseline value respectively.(3)The Claudin-2 m RNA decreased ratio of IRI group, mimic group, inhibitor group, mimic NC group and inhibitor NC group were 2.27±0.25, 4.25±0.63, 1.78±0.21, 2.31±0.29, 2.30±0.31 times of baseline value respectively, the Claudin-2 m RNA decreased ratio of IRI group, mimic group, and inhibitor group were significantly different, the value of mimic group was significantly lower than the inhibitor group(P < 0.05).(4)The Claudin- 2 protein expression level of IRI group, mimic group, inhibitor group, mimic NC group, inhibitor NC group were 0.694±0.072, 0.263±0.021, 0.860±0.080, 0.605±0.062, 0.524±0.052 respectively; the Claudin- 2 m RNA expression level of the IRI group, mimic group and inhibitor group were significantly different, the expression level of mimic group was significantly lower than the inhibitor group(P<0.05).(5)The CCK 8 OD value of IRI group, mimic group, inhibitor, mimic NC group, inhibitor NC group were 0.08±0.011, 0.03±0.012,0.13±0.030,0.08±0.010,0.09±0.015 respectively, the CCK 8 OD value of the IRI group, mimic group, and inhibitor groups were significantly different, the value of mimic group was significantly lower than the inhibitor group(P<0.05).(6) The cell apoptosis rate of IRI group, mimic group, inhibitor group, mimic NC group, inhibitor NC group were(64.2±2.80)%,(78.6±3.10)%,(48.2±2.38)%,(60.1±2.52)%,(66.8±2.91)% respectively, and the cell apoptosis rate of the IRI group, mimic group and inhibitor group were significantly different, the rate in mimic group was significantly higher than the inhibitor group(P<0.05). Conclusion During process of IRI, the mi RNA- 214 mediated the apoptosis of HK- 2 cell through an inhibition of Claudin- 2 m RNA and protein expression.