MicroRNA Expression Profiling In Male And Female Model Mice After Renal Ischemia-reperfusion Injury

Objective:To investigate the differential expression of micro RNAs in the kidney between female and male mice in order to study the effects and mechanisms of micro RNA in pathogenesis of ischemia-reperfusion injury between different genders.Methods:Nontrau-matic microvascular clamps were applied across both renal pedicles to construct male BALB/c mice model of renal ischemia-reperfusion injury. According the purpose of this experiment the mice were divided into four groups: T1 group BALB/C males blocking the renal vascular pedicle 45 min and 24 h reperfusion.T2 group BALB/C females blocking the renal vascular pedicle 45 min and 24 h reperfusion.C1 group BALB/C males which is only opened the abdominal cavity without blocking vascular renal pedicle. C2 group BALB/C females which is only open the abdominal cavity without blocking vascular renal pedicle.The mice were all feeded 24 hours after the surgery. Blood was collected to detect serum uria nitrogen and creatinine to determine kidney function 24 hours after the surgery. Then the mice were sacrificed eagerly,taking a surgical incision from Abdominal median incision of the model mouse to obtain bilateral kidneys, HE staining one kidney,optical microscope will be used to observed kidney histopathological changes.The other kidney will be used to detect the expression of micro RNA chip and put to other use.Results:(1) 24 hours after the surgery the survival rate of T1 group,T2 group,C1 group,C2 group were respective be 80%、100%、100%、100%. Serum creatinine and urea nitrogen in T2 group significantly lower than the T1 group, however these two groups were both higher than C1 group and C2 group(P<0.05). Serum creatinine and urea nitrogen in C1 group and C2 group have no significant difference(P>0.05).(2)Kidney histopathology score in group T2 is obviously lower than the T1 group,and both of them is obviously than C1 group and C2 group(P<0.05). Kidney histopathology between C1 group with C2 group have no significant difference(P>0.05).(3) Five micro RNAs were up-regulated between T2 and T1 groups. Twenty-nine micro RNAs differentially expressed in the T2 group and C2 group, including 25up-regulated micro RNAs and 4 down-regulated micro RNAs. Thirty-eight micro RNAs differentially expressed in T1 group and C1 group, including 9 up-regulated micro RNAs and 29 down-regulated micro RNAs. 102 micro RNAs differentially expressed in the C2 group and C1 group, including 22 up-regulated micro RNAs and80 down-regulated micro RNAs. Results suggested that there was differential expression in micro RNAs in the kidney before and after renal ischemia-reperfusion in male and female mice. These differentially expressed micro RNAs may be lead to different sensitivity and tolerance to the ischemia-reperfusion injury in the kidney of male and female mice.(4) comparison with the Micro RNA expression between four group BLBA/c mouse, The mir-21 and mir-664 both have significant difference,they expression in C1 group and C2 group have no significant difference,But mir-21 expression in T1 group is 1.5 times than C1 group,mir-21 expression in T2 group is 2 times than C1 group.the mir-664 expression in T1 group is 1.3 times than C1 group,mir-21 expression in T2 group is 2.2 times than C1 group. Real-time PCR results are in conformity with gene chip.Conclusion:(1) Renal ischemia reperfusion mode of BALB/c mice were Successfully constructed.(2) The susceptibility in the Renal ischemia reperfusion is difference between Gender differences.(3) The differential expression of micro RNAs in the kidney between female and male after renal ischemia reperfusion. The mir-21 and mir-664 both have significant difference.