| Objective: To explore the mechanism of the protective effect of The Centella Asiatica granules on renal ischemia reperfusion injury by PI3K/AKT/m TOR signaling pathway.Methods: Rats were randomly divided into 4 groups,10 rats in each group,which were sham operation group(Sham),model group(I/R),Centella Asiatica granules group(As)and Wortmannin group(Wort).The Sham group was gavaged with normal saline continuously for 40 days before the model and 3 days after the model.The Rats’ bilateral kidney pedicles were exposed for 45 minutes in the Sham group operation model.An equal volume of normal saline was injected 4 hours before modeling and 3 days after modeling,once every other day.In the I/R group,the bilateral renal pedicles were clamped with non-invasive arterial clips for 45 minutes.And then the renal blood flow is restored.The rest of the operation was the same as the Sham group.The As group was continuously gavaged with an aqueous solution of The Centella Asiatica granules40 days before modeling and 3 days after modeling.The rest operation of the As group was the same as the I/R group.The Wortmannin intervention was given to the Wort group 4 hours before modeling and 3 days after modeling,once every other day.The rest operation of the Wort group was the same as the As group.In this experiment,arterial blood was used to detect changes in renal function indexes of each group.The optical microscopy was used to observe and score changes in renal tissue morphology.The TUNEL method was used to detect renal cell apoptosis ratio in each group.The protein expression levels of PI3K/AKT/m TOR signaling pathway and CleavedCaspase-3 was detected by IHC.Results: After the establishment of renal IRI model,we check various indicators.(1)Kidney function: The Scr and Bun levels in the Sham group were low.The Scr and Bun levels in the I/R group were significantly higher than those in the Sham group(P<0.05).The levels in the As group were lower than those in the I/R group,which was still higher than those in the Sham group(P<0.05).The levels in the Wort group were significantly higher than those in the As group(P<0.05),without significant difference from the I/R group(P>0.05);(2)Pathological tissue score: Using artificial semi-quantitative scoring,the results showed that the pathological score of the Sham group was low,which was significantly lower than the I/R group(P<0.05).The pathological score of the As group was decreased compared with the I/R group,which was still higher than those in the Sham group(P<0.05).The score in the Wort group is higher than that in the As group(P<0.05),without significantly different from the I/R group(P>0.05);(3)Related indicators of renal cell apoptosis: The ratio of apoptotic cells and the expression of Cleaved-Caspase-3 were consistent in each group.The ratio of apoptotic cells and the expression of Cleaved-Caspase-3 in the Sham group were low,which were lower than that in the I/R group(P<0.05).The indexes of the As group was lower compared with the I/R group,which was still higher than those in the Sham group(P<0.05).The indexes in the Wort group is higher than that in the As group(P<0.05),without significantly different from the I/R group(P>0.05);(4)PI3K/AKT/m TOR signaling pathway expression levels:The signaling pathway expression levels in the Sham group were lower than that in the I/R group(P<0.05).The expression levels of Phospho-PI3 K,Phospho-AKT,Phospho-m TOR in the As group were significantly higher than those in the I/R group and Sham group,with significant differences(P<0.05);The expression levels in the Wort group were significantly lower than those in the As group but higher than those in the Sham group,with significant differences(P<0.05).There was no significant difference compared with the I/R group(P>0.05).Conclusions: The Centella Asiatica granules can improve renal ischemia reperfusion injury in rats,and may play a renal protective role by regulating the anti-apoptotic pathway of PI3K/AKT/m TOR signaling pathway. |
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