| Objective:Detectives the autophagy activity of PPAR gamma activates regulation of rat renal ischemia reperfusion injury of renal tubular epithelial cell.Methods:SPF grade SD male rats,200-250 g,The rats were divided into five groups.sham group: which is only opened the abdominal cavity without blocking vascular renal pedicle,closed the abdominal cavity after 45 min,DMSO was intragastric administration(2ml/kg*d)for one weeks before induction of ischemia.DMSO+IRI group(DIR group): which is used microvascular clamps blocks the renal vascular pedicle 45 min,the double renal blood flow was restored,and closed the abdominal cavity,DMSO was intragastric administration(2ml/kg*d)for one weeks before induction of ischemia.Pioglitazone+IRI group(PIR group): which is used microvascular clamps blocks the renal vascular pedicle 45 min,the double renal blood flow was restored,and closed the abdominal cavity,Pio was intragastric administration(10mg/kg*d)for one weeks before induction of ischemia.GW9662+IRI group(GIR group): which is used microvascular clamps blocks the renal vascular pedicle 45 min,the double renal blood flow was restored,and closed the abdominal cavity,GW9662 was administered intraperitoneally(1 mg/kg)24hours and 12 hours before induction of ischemia.Pioglitazone+GW9662+IRI group(PGIR): which is used microvascular clamps blocks the renal vascular pedicle45 min,the double renal blood flow was restored,and closed the abdominal cavity,Pio was intragastric administration(10mg/kg*d)for one weeks and GW9662 was administered intraperitoneally(1 mg/kg)24 hours and 12 hours before induction of ischemia.(n=5).The groups will be collect blood and obtain samples of rat kidney24 h after reperfusion,blood was collected to detect serum creatinine and urea nitrogen to determine renal function,and from rat abdominal incision to obtain kidneys,optical microscope wlli be used to observation the histopathologic changes by HE staining,the terminal deoxynucleotidyl transferase mediated nick end labeling method(TUNEL method)to detect the renal cell apoptosis of transfer,detect m RNA and protein expression of the PPAR gamma by RT-PCR and western blot,detect autophagy related protein LC3 and Beclin-1 expression changes of each groups by western blot.Results:(1)Blood serum creatinine and serum uria nitrogen: As compared with sham group,the IRI groups serum creatinine and serum uria nitrogen have significant elevation(P<0.05).As compared with DIR group,the PIR group erum creatinine and serum uria nitrogen ware significant descend(P<0.05),the GIR group erum creatinine and serum uria nitrogen have no statistically significance(P>0.05);As compared with PIR group,the PGIR group erum creatinine and serum uria nitrogen ware significant elevation(P<0.05).(2)HE staining: As compared with sham group,the IRI groups renal tissue pathological score was significantly elevation(P<0.05);As compared with DIR group,the PIR group pathological score was significant descend(P<0.05),the GIR group pathological score has no statistically significance(P>0.05);As compared with PIR group,the PGIR group pathological score was significant elevation(P<0.05).(3)The apoptosis of renal tubular epithelial cell: As compared with sham group,the number of cell apoptosis in the IRI groups was significant elevation(P<0.05);As compared with DIR group,the number of PIR group cell apoptosis was significantly descend(P<0.05),the number of GIR group cell apoptosis has no statistically significance(P>0.05);As compared with PIR group,the number of PGIR group cell apoptosis was significant elevation(P<0.05).(4)The PCR expression of PPAR gamma: As compared with sham group,the PIR groups and the PGIR group expression of PPAR gamma was significantly elevation(P<0.05),the DIR group expression of PPAR gamma has no statistically significance(P>0.05),the GIR group expression of PPAR gamma was descend(P<0.05);As compared with DIR group,the PIR group expression of PPAR gamma was significantly elevation(P<0.05);As compared with PIR group,the PGIR group expression of PPAR gamma ware significant descend(P<0.05).(5)The protein expression of PPAR gamma,LC3 and Beclin-1: As compared with sham group,the IRI groups expression of LC3 and Beclin-1 ware significantly elevation,but the DIR group expression of PPAR gamma has no tatistically significance(P>0.05),the PIR group and the PGIR group expression of PPAR gamma ware significantly elevation(P<0.05),the GIR group expression of PPAR gamma was significantly descend(P<0.05);As compared with DIR group,the PIR group expression of PPAR gamma,LC3 and Beclin-1 ware significantly elevation(P<0.05);As compared with PIR group,the PGIR group expression of PPAR gamma,LC3 and Beclin-1 ware significant descend(P<0.05).Conclusion:(1)Renal ischemia reperfusion can cause renal function and histopathological changes in rats,The expression of serum creatinine and blood urea nitrogen increased,and the apoptosis and necrosis of renal cells increased.(2)Pioglitazone activating PPAR gamma pathway is involved in protection of renal ischemia reperfusion injury.This effect is achieved by regulating the expression of LC3 and beclin-1. |
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