Mechanism Of Protect Effects Of Panax Notoginseng On Cisplatin-induced Acute Kidney Injury

Background: Acute kidney injury(AKI)is one of the common adverse effects of the chemotherapeutic drug cisplatin.Acute kidney injury(AKI)is one of the common adverse effects of the chemotherapeutic drug cisplatin,and there is still no recognized and stable drug available to treat cisplatin-induced acute kidney injury(CI-AKI).Panax ginseng(PN)is widely used in the treatment of many diseases as a herbal medicine to activate blood circulation and remove blood stasis.Some studies have shown that Panax ginseng can improve acute kidney injury by alleviating.However,the beneficial effects of Panax ginseng on endoplasmic reticulum stress and mitochondrial dysfunction in cisplatin-induced acute kidney injury remain to be investigated.Objective: To investigate the role and molecular mechanism of Panax notoginseng(PN)in the prevention and treatment of Cisplatin-induced Acute Kidney Injury(CI-AKI),and to provide new ideas for the prevention and treatment of CI-AKI.Methods:1.We constructed an animal model of cisplatin-induced acute kidney injury in C57BL/6 mice,which were pretreated with Panax ginseng for 7 days before modeling.Using kits and ELSA kits to detect blood creatinine,blood urea nitrogen,cystatin C,glutamic oxalacetic transaminase and glutamic alanine transaminase in mice;HE staining and PAS staining were performed on mouse kidney tissue sections to assess pathological injury;kidney tissues were homogenized and then apoptosis-related proteins such as Bcl-2,Bax and cleaved-caspase-3 were measured;analysis of the expression levels of endoplasmic reticulum-related proteins such as PERK,ATF6 and CHOP were analyzed;the ultrastructure and lesions of mouse mitochondria were observed by electron microscopy,and the levels of mitochondrial ATP,reactive oxygen species and mitochondrial membrane potential in mouse kidney were measured by using ATP kit,MitoSOX kit and JC-1 kit;the expression levels of mitochondrial splitting proteins Drp1,Fis1 were determined;and XBP1 and Mfn2 expression in mouse kidney was detected by immunohistochemistry,and XBP1-Mfn2 signaling pathway protein expression level was further detected by Western blot.2.We constructed a cisplatin-induced injury model of human renal proximal tubular epithelial cells(HK-2),and pretreated by adding Panax ginseng 1 hour before modeling.Flow cytometry was used to detect the apoptosis level.And apoptosis-related proteins such as Bcl-2,Bax and cleaved-caspase-3 were measured;analysis of the expression levels of endoplasmic reticulum-related proteins such as PERK,ATF6 and CHOP were analyzed;the ultrastructure and lesions of mouse mitochondria were observed by electron microscopy,and the levels of mitochondrial ATP,reactive oxygen species and mitochondrial membrane potential in mouse kidney were measured by using ATP kit,MitoSOX kit and JC-1 kit;the expression levels of mitochondrial splitting proteins Drp1,Fis1 were determined;and XBP1-Mfn2 signaling pathway protein expression level was further detected by Western blot.Results:1.PN pretreatment improved blood creatinine,blood urea nitrogen,cystatin C,and glutamic oxalacetic transaminase and glutamic alanine transaminase levels in mice with cisplatin-induced acute kidney injury.Panax ginseng pretreatment also attenuated cisplatin-induced renal histopathological injury in mice.It attenuated renal tubular apoptosis and regulated PERK,ATF6 and CHOP,the signature proteins of endoplasmic reticulum stress.It increased ATP energy metabolism,improved mitochondrial morphology and mitochondrial superoxide levels in renal tubular cells,restored cellular mitochondrial membrane potential levels,restored mitochondrial division-related protein Drp1 and Fis1 expression levels,and regulated XBP1-Mfn2 pathway protein expression levels.2.PN reduced cisplatin-induced apoptosis in HK-2 cells and improved PERK,ATF6 and CHOP,the signature proteins of endoplasmic reticulum stress.It increased ATP energy metabolism and attenuated cellular mitochondrial superoxide levels and cellular mitochondrial membrane potential levels,restored the expression levels of mitochondrial division-related proteins Drp1 and Fis1,and regulated the XBP1-Mfn2 pathway protein expression levels.Conclusion: PN alleviated renal tubular injury in CI-AKI by improving mitochondrial dynamics and mitochondrial function as well as activating XBP1-Mfn2 pathway.