Study On The Mechanism Of Yishen Xiezhuo Formula In Ameliorating Cisplatin-Induced Acute Kidney Injury By Attenuating Renal Tubular Cell Premature Senescence

Objective:Acute kidney injury(AKI),which is prevalent,challenging to treat,and has a poor prognosis,has emerged as a pressing global public health issue.Although cisplatin(DDP)is a common antineoplastic drug,the incidence of AKI is rising with clinical use,indicating the need for more clinical research into the pathogenesis of AKI brought on by cisplatin and for effective treatment options.Senescence of renal tubular epithelial cells is a significant factor in cisplatin-induced AKI,but the precise mechanism is still unclear.Previous research has shown that the Yishen Xiezhuo formula can improve the prognosis of patients with cisplatin-induced AKI,but the microbiological mechanisms of action are still not fully understood.Therefore,the aim of this study is to initially explore whether YSXZ can reduce renal tubular epithelial cell senescence and ameliorate cisplatin-induced AKI,and to provide a theoretical basis for the clinical application of YSXZ.Methods:1.An HCC tumor-bearing mouse model was developed and randomly divided into groups.According to the grouping circumstances,intraperitoneal injection of cisplatin,intragastric administration of YSXZ,and intragastric administration of NDQ medication solution were employed.The mice’s overall condition was checked,and their body weight and tumor volume were recorded.Each group’s tumor inhibition rate and spleen index(Spleen Index,SI)were calculated.Kits were used to measure the levels of creatinine and blood urea nitrogen in mice serum;pathological damage to mice kidney tissue was examined and assessed using H&E and PAS staining.To detect the senescence of the mice’s renal cortex,use senescence staining and perform semiquantitative statistical analysis.2.Cell experiment:Human proximal renal tubular epithelial cells(HKC-8)were used as experimental subjects to examine the effects of YSXZ drug-containing serum on cellular senescence staining characteristics,cellular senescence-related protein expression levels,cell cycle and cellular senescence-related secretory phenotypes IL-1 and IL-6 after cisplatin intervention using SA-β-gal senescence staining,Western Blot,Flow cytometry,ELISA,etc.3.Utilize the TCMSP database to gather and filter YSXZ active components.To get the core targets of YSXZ for the treatment of AKI,we mined and collected possible AKI targets in major illness databases,as well as mapped the active ingredient targets of YSXZ and AKI-related targets.Submit the core targets to the STRING database in order to build a protein-protein interaction(PPI)network and analyze core modules.The major pathways of YSXZ in the treatment of AKI were produced by doing GO and KEGG enrichment analysis on the core targets using the Metascape database.Lastly,Western Blot was used to verify the main pathways.Results:1.The overall condition of HCC tumor-bearing mice after cisplatin therapy was poor,and their body weight and spleen index were considerably lower than those in the control group(P<0.0001,P<0.05).The body weight and spleen index of mice treated with YSXZ in combination with cisplatin were greater than those of mice treated with cisplatin alone(P<0.05,P<0.05),but there was no significant difference in average tumor volume or tumor inhibition rate.Simultaneously,YSXZ coupled with cisplatin therapy can minimize the renal tubular injury caused by cisplatin(P<0.0001)and postpone the rising trend of creatinine and blood urea nitrogen.Yishen Xiezhuo Formula also reduced the senescence staining positive area of the HCC tumor bearing mice’s kidney tissue aging and delayed cisplatin-induced renal injury.2.SA-β-gal staining demonstrated that senescent cells grew considerably(P<0.001)in the DDP group,while 5%and 10%YSXZ treatments reduced the positive staining area of SA-β-gal(P<0.001,P<0.001).Simultaneously,the expression of aging-related proteins was significantly up-regulated in the DDP group,with statistical differences(P<0.01,P<0.01,P<0.05,P<0.05).The expression of associated proteins was strongly down-regulated after YSXZ intervention,and the impact of 10%YSXZ was more pronounced.Moreover,cells in the DDP group had S-phase cell cycle arrest(P<0.001),and YSXZ intervention may restore the S-phase arrest produced by cisplatin and diminish the secretion of aging-related secretory phenotypes IL-1 and IL-6.3.TCMSP was used to extract the active components and targets of YSXZ,and after merging and eliminating duplicate data,62 active ingredients and 219 targets of YSXZ were retrieved.After combining and eliminating duplicate records,we acquired 12,647 acute renal injury-related targets from major disease databases.The screened YSXZ active ingredient targets and AKI targets were intersected,yielding 214 core YSXZ targets for treating AKI.The core targets were subjected to PPI network design as well as GO and KEGG analysis,and it was discovered that the MAPK signaling pathway was closely linked.Conclusions:1.Stress induced premature senescence of renal tubular epithelial cells is an important mechanism in the development of CI-AKI.2.The mechanism of Yishen Xiezhuo Formula in the treatment of CI-AKI may be related to the inhibition of MAPK pathway-induced stress induced premature senescence of renal tubular epithelial cells.