Mechanism of EGCG Regulating PI3K/Akt Pathway on CIS-induced Acute Kidney Injury in Rats
[Objective]This study was aimed to investigate the protective effect and mechanism of epigallocatechin-3-gallate(EGCG)on acute kindey injury(AKI)induced by cisplatin(CIS)in rats,so as to provide experimental basis for the clinical use of CIS and EGCG.[Method]Forty male Wistar rats were randomly divided into 5 groups.Rats in CON and CIS groups were perfused with normal saline,and rats in EGCG,CE and inhibitor groups were gavaged with 40 mg/kg EGCG for 28 consecutive days.On the 26th day,rats in CIS,CE and inhibitor groups were intraperitoneally injected with 7 mg/kg CIS,rats in CON and EGCG groups were injected with an equal volume normal saline,and rats in inhibitor group was intraperitoneally injected with 5 mg/kg LY294002 for 3 consecutive days.The kindey samples of all rats were collected on the 29th day.HE staining and transmission electron microscope were used to observe the changes of renal pathology and cellular mitochondrial function,Western blotting was used to detect the expression of autophagy proteins(LC3 Ⅱ/Ⅰ,ATG5,ATG12,Beclin-1 and p62)and pathway proteins(PI3K,Akt,p-PI3K and p-Akt).Immunofluorescence was used to detect the expression of autophagy-related proteins LC3Ⅱ/Ⅰ and p62 in kindey of rats.[Result]The results of HE staining and transmission electron microscope showed that pretreatment of EGCG alleviated the necrosis and exfoliation of renal tubular epithelial cells,infiltration of inflammatory cells,swelling and degeneration of cell mitochondria and rupture of mitochondrial crest induced by CIS.Western blotting results showed that compared with CON group,the expressions of LC3 Ⅱ/Ⅰ,ATG5,ATG12 and Beclin-1 protein in kidney of CIS group were extremely significantly increased(P＜0.01),while the expressions of p62,p-PI3K/PI3K and p-Akt/Akt were extremely significantly decreased(P＜0.01),but this phenomenon was reversed in EGCG pretreatment group.Immunofluorescence results showed that compared with CE group,the pathological damage in inhibitor group was aggravated,the fluorescence signal of LC3 Ⅱ/Ⅰ was enhanced,and p62 was weakened.[Conclusion]Pretreatment of EGCG could inhibit autophagy by activating PI3K/Akt signaling pathway,thus alleviating CIS-induced AKI in rats,and this protective effect could be offset by PI3K/Akt signaling pathway inhibitor LY294002.
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