| Objective: To investigate the role of neutrophil extracellular traps(NETs)in renal ischemia-reperfusion injury and to clarify the protective effect of DNase-1 on renal ischemia-reperfusion injury.Methods: Eighteen SPF wistar male rats were weighed and divided into three groups according to the random number method: control group: six rats underwent right nephrectomy,and the left kidney was not subjected to ischemia-reperfusion treatment;IR group(renal ischemia-reperfusion group): After completion of right nephrectomy,six rats were subjected to ischemia for 45 minutes after occlusion of the left renal artery with an arterial clamp,and then the kidney was reperfused for 48hours;IR+DNase-1 group(renal ischemia-reperfusion+DNase-1 intervention group):After completion of right nephrectomy,six rats were subjected to ischemia for 45 minutes by blocking the left renal artery with an arterial clamp.We injected DNase-1solution(usage: 0.1 mg/kg)intraperitoneally 15 minutes before renal restoration of blood perfusion.We then opened the arterial clamp to allow the kidney to be reperfused for 48 hours.Renal function impairment was assessed by detecting Scr and BUN in rat serum.HE staining was used to observe the renal tissue injury,and the expression of MPO and Cit-H3 in rat renal tissue was detected by immunohistochemistry to investigate the production of NETs.Bax mRNA and BCL-2mRNA expression in renal tissue were measured by RT-qPCR to assess the degree of apoptosis.Result:(1)Assessment of renal function revealed that there were significant differences in Scr and BUN levels between the control group,the IR group and the IR+DNase-1 group.The level limit of Scr and BUN of rats in the IR+DNase-1 group was significantly decreased compared with the IR group,which was statistically significant(P<0.05);(2)HE staining of renal tissue revealed that compared with the IR group,the IR + DNase-1 group showed a reduction in the degree of tubular injury,congestion and edema and necrosis and other injuries.The results of pathological score showed that the score of IR group was significantly higher than that of control group(P < 0.05),while the score of IR+DNase-1 group was significantly lower than that of IR group(P < 0.05);(3)Compared with the control group,the serum SODactivity of the rats in the IR group was significantly decreased,with statistical significance(P < 0.05).However,the SOD activity of rat serum in the IR+DNase-1group was increased compared with the IR group(P<0.05),and the difference was statistically significant;(4)Immunohistochemical staining results revealed that MPO and Cit-H3 contents in the kidney tissue of the IR group were significantly higher than those of the control group(P< 0.05),and the expression of the IR+DNase-1group was decreased compared with the IR group(P < 0.05);(5)RT-qPCR revealed that the BAX mRNA was significantly increased and the Bcl-2 mRNA level was significantly decreased in the IR and the IR+DNase-1 group compared with the control group,and the difference was statistically significant(P < 0.05).Compared with the IR group,the BAX mRNA of the serum of rats in the IR+DNase-1 group was significantly decreased,and the Bcl-2 mRNA was significantly increased,and the difference was statistically significant(P < 0.05).Conclusion:(1)In the ischemia-reperfusion injury model,the content of NETs in renal tissue was significantly increased,and DNase-1 could reduce the content of NETs in the kidney during renal ischemia-reperfusion injury;(2)DNase-1pretreatment significantly reduced renal injury in ischemia-reperfusion of the kidney;(3)DNase-1 preconditioning may attenuate renal ischemia-reperfusion injury by inhibiting apoptosis. |
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