Target Regulation Of MiR-218 To Claudin-2 And Its Role In Ischemia Reperfusion Injury Of HK-2 Cell

ObjectiveTo explore the target regulation of miR-218 to Claudin-2 and its role in ischemia reperfusion injury of Renal Tubular Epithelial(HK-2) Cell.MethodsHK-2 cells as the research object, antimycin A,A23187 and 2-deoxy-D-glucose were used to establish ischemia-reperfusion injury as an in vitro cell modelThe first part: the experiment is divided into the following groups:(1)control group,(2) I/R group(subgroups: 0h, 8h, 12 h, 24 h, 48 h after reperfusion), To study the dynamic changes of cell apoptosis, cell viability, miR-218 and Claudin-2 in HK-2 cells during ischemia-reperfusion injury.The second part: induced IRI after transfection with miR-218 mimics, miR-218 inhibitor and meaningless miR,experiment index measured in 12 h after reperfusion, experiment groups divide into:(1)I/R-12 h group,(2) mimics group,(3) inhibitor group,(4) mimics-NC group,(5)inhibitor-NC group. To study the cell apoptosis, cell viability and Claudin-2 expression.ResultsPart one :During the ischemia-reperfusion injury in HK- 2 cells, in I/R-12 h,apoptotic rate [(63.24±4.29)%] was highest, cell viability(0.11±0.03) was lowest(P<0.05); The expression level of miR-218 was closely related to the cell apoptotic rate and cell viability, in I/R-12 h,it was 6.44 times as high as that of the baseline(P<0.05); The expression level of cldn-2 m RNA and protein content was parallel to the level of miR-218 expression, in I/R-12 h, cldn-2 m RNA was lowest, decline(3.06 + 0.16) times, the relative expression level of cldn-2 protein content fell to the lowest point(0.13±0.01)(P<0.05).Part two : After transfection, the apoptotic rate and expression of miR-218 in mimics group were significantly increased, 1450 times as high as that of the baseline, compared with I/R-12 h group, the apoptotic rate[( 80.18±4.23) %] were significantly increased, cell viability(0.05±0.01)were significantly decreased(P<0.05), expression of Claudin-2 m RNA [decline(3.62±0.14) times] and protein content(0.09±0.01)were significantly decreased(P<0.05). Compared with I/R-12 h group, the apoptotic rate [(51.22±3.21)%] in inhibitor group were decreased, cell viability were increased(0.18±0.02)(P<0.05), expression of Claudin-2 m RNA [decline(1.59±0.03) times] and protein content(0.29±0.02)were significantly increased(P<0.05).ConclusionMiR-218 targeted inhibit the m RNA and protein expression of cldn-2,mediate HK-2 cells ischemia-reperfusion injury, inhibit MiR-218 can reduce HK-2 cells ischemia-reperfusion injury.