| Objective:To observe the effect of pirinixic acid and ischemic proconditioning to treat the rats with renal ischemia reperfusion injury.Methods:Part One:Fifty rats were divided randomly into five groups:Sham group (S), I/R for20minutes group (IR20), I/R for40minutes group (IR40), I/R for60minutes group (IR60), and I/R for80minutes group (IR80). Experiment steps:Resected the right kidney, but did not occlude the left renal artery in S group,(n=10). IR20group (n=10):resected the right renal, then occluded the left renal artery for20minutes to cause ischemia. IR40group:resected the right renal, then occluded the left renal artery for40minutes to cause ischemia. IR60group:resected the right renal, then occluded the left renal artery for60minutes to cause ischemia. IR80group:resected the right renal, then occluded the left renal artery for80minutes to cause ischemia.24hours after the operation,the left kidney and blood of rats were collected in all groups. Used automatic biochemistry analyzer to measure the levels of serum urea nitrogen and creatinine.Part Two:Ninety rats were divided randomly into five groups:Sham group(S), I/R group (IR), Pirinixic acid group1(PA1), Pirinixic acid group5(PA5), Pirinixic acid group10(PA10),. Experiment steps:Resected the right kidney, but did not occlude the left renal artery in S group,(n=18). IR group (n=18):resected the right renal, then occluded the left renal artery for60minutes to cause ischemia. PA1group(n=18):thirty minutes before operation, pirinixic acid lmg/kg were injected into abdominal cavity of rats, and other steps were same to the IR group; PA5group(n=18):thirty minutes before operation, pirinixic acid5mg/kg were injected into abdominal cavity of rats, and other steps were same to the IR group; PA10 group(n=18):thirty minutes before operation, pirinixic acid10mg/kg were injected into abdominal cavity of rats, and other steps were same to the IR group.Four hours after the operation,the left kidney and blood of rats were collected in all groups. Used automatic biochemistry analyzer to measure the levels of serum urea nitrogen and creatinine; Used thibabituric acid to measure the levels of malonaldehyde; Used xanthine oxidase to measure the levels of superoxide dismutase.Part Three:Ninety rats were divided randomly into five groups:Sham group(S), I/R group (IR), Ischemic proconditioning group1(IPO1), Ischemic proconditioning group2(IPO2), Ischemic proconditioning group3(IPO3). Experiment steps:Resected the right kidney, but did not occlude the left renal artery in S group,(n=18). IR group (n=18):resected the right renal, then occluded the left renal artery for60minutes to cause ischemia. IPO1group (n=18):Resected the right kidney of rats, and occluded the left renal artery for sixty minutes, then occluded the left renal artery for thirty seconds after reperfused for thirty seconds for one time before reperfusion; IPO2group (n=18): Resected the right kidney of rats, and occluded the left renal artery for sixty minutes, then occluded the left renal artery for fifteen seconds after reperfused for fifteen seconds for two times before reperfusion; IPO3group (n=18):Resected the right kidney of rats, and occluded the left renal artery for sixty minutes, then occluded the left renal artery for ten seconds after reperfused for ten seconds for three times before reperfusion;.Four hours after the operation,the left kidney and blood of rats were collected in all groups-Used automatic biochemistry analyzer to measure the levels of serum urea nitrogen and creatinine; Used thibabituric acid to measure the levels of malonaldehyde; Used xanthine oxidase to measure the levels of superoxide dismutase.Part Four:Ninety rats were divided randomly into five groups:Sham group(S), I/R group (IR), Ischemic proconditioning group (IPO), Pirinixic acid group(PA), Pirinixic acid and Ischemic proconditioning group (PAIPO). Experiment steps: Resected the right kidney, but did not occlude the left renal artery in S group,(n=18). IR group (n=18):resected the right renal, then occluded the left renal artery for60 minutes to cause ischemia. IPO group (n=18):Resected the right kidney of rats, and occluded the left renal artery for sixty minutes, then occluded the left renal artery for ten seconds after reperfused for ten seconds for three times before reperfusion; PA group(n=18):thirty minutes before operation, pirinixic acid5mg/kg were injected into abdominal cavity of rats, and other steps were same to the IR group; PAIPO group (n=18):pirinixic acid5mg/kg were injected into abdominal cavity of rats, thirty minutes before operation, whose other steps were same to the IPO group. Four hours after the operation,the left kidney and blood of rats were collected in all groups. The pathological changes of the renal were checked by light microscope with HE staining. Used automatic biochemistry analyzer to measure the levels of serum urea nitrogen and creatinine; Used thibabituric acid to measure the levels of malonaldehyde; Used xanthine oxidase to measure the levels of superoxide dismutase; Used ELISA to measure the levels of TNF-α and IL-1β; Used the immunohistochemistry methods to mesure the expression of intercellular adhesion molecule-1of the kidney.Results1. Did not found obviously abnormality in the S group; The injury in the left kidney was obvious in the IR group; Compared to the IR group, the injury in the left kidney of IPO group, PA group and PAIPO group wre lighter.2. The levels of serum urea nitrogen and creatinine in the IR group were higher than the S group (P<0.05), and the levels of serum urea nitrogen and creatinine in the IPO group, PA group and PAIPO group were lower than the IR group(P<0.05); The levels of serum urea nitrogen and creatinine in the IPO group and PA group were higher than the PAIPO group (P<0.05).3. The levels of MDA, TNF-α and IL-1β in the IR group were higher than the S group (P<0.05), and the levels of MDA, TNF-α and IL-1β in the IPO group, PA group and PAIPO group were lower than the IR group(P<0.05); The levels of MDA, TNF-α and IL-1β in the IPO group and PA group were higher than the PAIPO group (P<0.05).4. The levels of SOD in the IR group were lower than the S group (P<0.05), and the levels of SOD in the IPO group, PA group and PAIPO group were higher than the IR group(P<0.05); The levels of SOD in the IPO group and PA group were lower than the PAIPO group (P<0.05).5. ICAM-1did not obviously express in the S group; The levels of ICAM-1expression were higher in the IR group; The levels of ICAM-1expression were lower in IPO group and PA group; In PAIPO group, the levels of ICAM-1espression were lower than the IPO group and PA group. The mean optical density value in the IPO group and PA group were higher than the S group and PAIPO group (P<0.05), but lower than IR group(P<0.05); Compared with IPO group, the mean optical density value in PA group that had no distinctly difference with the IPO group (P>0.05).Conclusions1. The rats with ischemia for60minutes followed by reperfusion could cause renal ischemical-reperfusion injury.2. Both pirinixic acid and ischemic proconditioning could protect the kidney from ischemia reperfusion injury, perhaps they proctected the kidney by lift SOD capability and lower MDA, TNF-α IL-1β, ICAM-1levels.3. Pirinixic acid and ischemic proconditioning could cooperate in the protection to the kidney from ischemia injury reperfusion. |
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