| Objective: Renal ischemia-reperfusion injury is a common injury in both medicine and surgery.HMGB1 antibody has protective effect on renal ischemia-reperfusion injury,but its expression relationship on pathological level is lack of research,and compared with other protective agents of renal ischemia-reperfusion injury,there is also lack of comparative analysis.Therefore,we established an experimental model of renal ischemia-reperfusion injury in rats,observed the renal injury in different treatment methods and different treatment time,and then discussed how HMGB1 antibody can protect the renal ischemia-reperfusion injury,and compared with ethyl pyruvate.Methods: the rat model of renal ischemia-reperfusion injury was established.They were randomly divided into four groups: blank group,ischemia-reperfusion group,ethyl pyruvate group and HMGB1 antibody group.Each group was divided into four different time groups according to 5 minutes,15 minutes,30 minutes and 40 minutes.(1)Blank(sham)group: the rats were injected with normal saline 30 minutes before operation,the posterior abdominal cavity was opened,the right renal pedicle was exposed for the corresponding time,and then the incision was closed layer by layer.24 hours later,the right kidney and abdominal aorta of the rats were taken from the abdominal cavity.(2)In the group ofischemia-reperfusion(IRI),normal saline was injected intraperitoneally30 minutes before the operation,and the hind abdominal cavity of rats was opened.The right renal arteriovenous and ureter were clamped with noninvasive microvascular respectively,and then the blocking was released.The incision was closed layer by layer.24 hours later,the hind abdominal cavity of rats was opened to collect the right renal specimen and abdominal aorta blood specimen.(3)In the group of ethyl pyruvate,the rats were injected with ethyl pyruvate(40 mg / kg)30 minutes before operation.After opening the abdominal cavity of the rats,the right renal artery and vein and ureter were clamped with noninvasive microvascular respectively for the corresponding time,then the blocking was relieved and the incision was closed layer by layer.24 hours later,the right kidney and abdominal aorta of the rats were taken from the abdominal cavity.(4)In the HMGB1 antibody group,the rats were injected with HMGB-1antibody(30mg / kg)intraperitoneally 30 minutes before operation,and then the right renal artery and vein and ureter were clamped with noninvasive microvascular respectively for the corresponding time to release the blocking,and the incision was closed layer by layer.24 hours later,the right kidney and abdominal aorta of the rats were taken from the abdominal cavity.Immunohistochemistry,TUNEL and he staining were used to detect the percentage of renal cell apoptosis,serum creatinine level in the blood of abdominal aorta;Results: 1.Immunohistochemistry results: there was no significant difference in the expression of HMGB-1 between different treatment time in the blank group.When the treatment measures were ischemia-reperfusion,ischemia-reperfusion plus ethyl pyruvate and ischemia-reperfusion plus HMGB-1 antibody,the expression of HMGB-1in different treatment time was not completely equal,the difference was statistically significant Meaning(P < 0.05).The results showed that the longer the treatment time was,the higher the expression of HMGB-1(mean optical density)was.There was no significant difference between5 min and 15 min,and the difference between the other treatment time was statistically significant(P < 0.001).The longer the treatment time was,the expression of HMGB-1(mean optical density)was in ethyl pyruvate group The higher the density was,the higher the expression of HMGB-1between 5min and 15 min,15min and 30min(mean optical density)had no statistical significance,and the difference between the other treatment time had statistical significance(P < 0.05);the overall performance of HMGB-1 antibody group was that the longer the treatment time,the higher the expression of HMGB-1(mean optical density),and the HMGB-1 immunohistochemical table between 5min,15 min and 30 min There was no significant difference in the average optical density(P <0.05).2.TUNEL renal cell apoptosis percentage results: when the treatment time was 5 minutes,there was no significant difference inTUNEL renal cell apoptosis percentage among the four treatment measures.When the treatment time was 15 minutes,30 minutes and 40 minutes,the percentage of TUNEL renal cell apoptosis among the four treatment measures was not completely equal,the difference was statistically significant(P < 0.05).The results showed that the apoptosis percentage of TUNEL kidney cells in the ischemia-reperfusion group and the HMGB-1 antibody group was significantly higher than that in the blank group(P < 0.05),and there was no significant difference between the ischemia-reperfusion group,the ethyl pyruvate group and the HMGB-1 antibody group;when the treatment time was 30 min,the apoptosis percentage of TUNEL kidney cells in the ischemia-reperfusion group and the HMGB-1 antibody group was higher than that in the blank group(P < 0.05)The apoptosis percentage of TUNEL renal cells in the three groups of ethyl group and HMGB-1 antibody group was higher than that in the blank group(P < 0.05).There was no significant difference between the ischemia-reperfusion group and the pyruvate ethyl group,the pyruvate ethyl group and HMGB-1 antibody group.When the treatment time was 40 minutes,the apoptosis percentage of TUNEL renal cells in the blank group,the ischemia-reperfusion group,the pyruvate ethyl group and the HMGB-1 antibody group were two The differences were statistically significant(P < 0.001),I / R group > ethyl pyruvate group >HMGB-1 antibody group > blank group.3.The results of creatinine valuetest: when the treatment time was 5min,15 min and 30 min,there was no significant difference in serum creatinine value between the four treatment measures.When the treatment time was 40 min,the serum creatinine value between the four treatment measures was not completely equal,the difference was statistically significant(P < 0.001).The results showed that when the treatment time was 40 minutes,the serum creatinine values of blank group,ischemia-reperfusion group,ethyl pyruvate group and HMGB-1 antibody group were all statistically significant(P < 0.001),ischemia-reperfusion group > ethyl pyruvate group > HMGB-1 antibody group > blank group.4.He microscopic examination results: for the ischemia-reperfusion group,ethyl pyruvate group and HMGB1 antibody group,the difference between the three groups was not significant at 5 min and 15 min.Under the light microscope,we can see that part of the glomerulus is edematous,the volume is enlarged,the basement membrane of capillaries is thickened,some of the epithelial cells of renal tubules are vacuolated and degenerated,and vacuoles and granular substances of different sizes and quantities can be seen in the cytoplasm.At 30 minutes of ischemia-reperfusion,ethyl pyruvate and HMGB1 antibody,we observed the deposition of flocculent protein like substances and scattered cell fragments in the lumen,and congestion and congestion in interstitial vessels.The vacuolar degeneration and glomerular edema of renal tubularepithelial cells increased significantly compared with 5 and 15 minutes.There was no significant difference among the three groups in HE staining microscope.After 40 minutes of ischemia,the damage of renal tissue was further aggravated.We can see that there were congestion and congestion in the interstitial vessels in different degrees,local perivascular edema and pink edema fluid.The pathological changes of renal tissue in HMGB1 antibody group were better than those in ethyl pyruvate group,and the pathological changes in ethyl pyruvate group were better than those in ischemia-reperfusion group.Conclusion: 1.HMGB1 antibody can reduce the expression of HMGB1 protein in renal ischemia-reperfusion injury,reduce the pathological changes of renal morphology and structure,and protect the function of kidney.2、 When the time of renal ischemia reaches 30 min and 40 min,early administration of HMGB1 antibody can significantly reduce the renal ischemia-reperfusion injury.3.When the time of renal ischemia is more than 30 minutes,the effect of HMGB1 antibody on renal ischemia-reperfusion injury is better than that of ethyl pyruvate. |
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