Protective Effect Of Aspirin Against Cisplatin-induced Nephrotoxicity In Mice

Cisplatin（CIS）is a clinical chemotherapeutic drug widely used in solid malignant tumors such as lung,testicular and ovarian cancer,but the production of nephrotoxicity limits the application of this type of drug.According to statistics,25%to 30%of patients receiving cisplatin chemotherapy have acute kidney injury（Acute kidney injury,AKI）.Therefore,it is the focus of attention and research to find drugs and methods that can reduce the nephrotoxicity of cisplatin without affecting the anti-tumor effect of cisplatin and improve the quality of cisplatin chemotherapy.Studies have shown that aspirin can reduce the nephrotoxicity caused by cisplatin,but the protective mechanism is not yet clear.In addition,aspirin can also inhibit the proliferation and spread of tumor cells and reduce the resistance of tumor cells to chemotherapeutic drugs.This study intends to explore the protective mechanism of aspirin on cisplatin-induced acute kidney injury and the alleviation of cisplatin resistance.The research is of great significance.In the experiment,60 male Kunming mice were random Ly divided into 6 groups,namely the blank control group,the cisplatin administration group,and the 5/10/20/40 mg/kg aspirin+cisplatin group.The study first passed the mouse serum Biochemical index detection and histopathological observation explore the physiological state of mice and the optimal dose of aspirin protection.Secondly,the pro-inflammatory factors（TNF-α,NF-κB,IL-1β,IL-6）,apoptosis-related indicators（BAX,Bcl-2,Caspase3）,oxidative stress indicators（T-AOC,CAT,SOD,GSH,NO,MDA,ROS）and mitochondrial function indicators（mt DNA,ATP,ATPase activity,mitochondrial respiratory chain complex enzymes ND1,Atp5b,SDHD,etc.）expression in serum and kidney tissues were detected by ELISA kit,q RT-PCR and Western blot.Finally,the effects of aspirin combined with cisplatin on the viability and proliferation of the cisplatin-resistant cell line CHMp ^CISwere tested by CCK-8 and plate clone formation.The specific experimental results are as follows:（1）Different doses of aspirin have different effects on cisplatin-induced acute kidney injury mouse models,and the best dose of aspirin to exert a protective effect is 10mg/kg.（2）By detecting the expression of pro-inflammatory factors in the mouse kidney group,it was found that compared with the control group,the relative expression of pro-inflammatory factors TNF-α,NF-κB,IL-1βand IL-6 was increased in the cisplatin group.Under the action of aspirin,the expression levels of pro-inflammatory factor genes and proteins were significantly reduced,indicating that aspirin can reduce the inflammatory response induced by cisplatin.（3）Cisplatin induced increased ROS,increased NO and MDA content,decreased T-AOC,decreased CAT and SOD activity,and decreased GSH content in mouse kidney tissue.Under the action of aspirin,the content of ROS,NO and MDA was significantly reduced,the activity of CAT and SOD was increased,and the content of T-AOC and GSH was significantly increased,indicating that aspirin can alleviate the oxidative stress induced by cisplatin.（4）Transmission electron microscopy showed that the mitochondria in the kidney tissues of the cisplatin group mice were swollen and broken,mt DNA levels decreased,ATP content decreased,ATPase activity decreased,mitochondrial respiratory chain enzyme-related gene expression decreased,and apoptosis-promoting genes BAX and p-Caspase3 expression decreased Increased,the expression of anti-apoptotic gene Bcl-2 decreased.Under the action of aspirin,mitochondrial swelling was reduced,mt DNA level rose,ATP content increased,ATPase activity increased,mitochondrial respiratory chain enzyme-related gene expression increased to varying degrees,and apoptosis-promoting genes The expression of BAX and p-Caspase3 decreased,and the expression of anti-apoptotic gene Bcl-2 increased.Shows that aspirin can affect ATP content and ATPase activity,reduce mitochondrial dysfunction,and reduce cisplatin-induced apoptosis.（5）The results of detection of AMPK-PGC-1αpathway related genes in mouse kidney tissue showed that the expressions of p-AMPK and mitochondrial related genes PGC-1α,NRF1 and TFAM in the cisplatin group were significantly lower than those in the control group,And the expressions of p-AMPK,PGC-1α,NRF1 and TFAM in the cisplatin+aspirin group were higher than those in the cisplatin group.This indicates that aspirin may regulate mitochondrial production by activating the AMPK-PGC-1αpathway,and alleviate the acute kidney injury induced by cisplatin.（6）The results of cell experiments showed that the effect of cisplatin on CHMp ^CIS caused a slight decrease in cell activity and proliferation ability.After aspirin was added,the cell activity and proliferation ability of CHMp ^CIS decreased significantly and depended on the dose of cisplatin.It is suggested that aspirin can not only strengthen the anti-tumor effect of cisplatin,but also restore the sensitivity of chemotherapy drugs.In summary,10mg/kg aspirin protects the body from acute kidney injury by alleviating cisplatin-induced inflammatory response,oxidative stress,mitochondrial dysfunction and apoptosis.Further studies have shown that the protective effect of aspirin is related to the activation of AMPK-PGC-1αpathway.In addition,this study also found that aspirin can restore the drug sensitivity of cisplatin-resistant cells.This not only provides theoretical support for the protective function of aspirin on AKI,but also provides possible prevention and treatment methods for alleviating the resistance of cisplatin and other chemotherapeutic drugs,which is of great significance to improve the survival rate of patients.