Study On The Mechanism Of Zhibai Dihuang Granule On Cisplatin Induced Acute Renal Injure Based On Network Pharmacology

Cisplatin is a non-specific chemotherapy drug,but cisplatin acute renal injury is a dose limiting toxicity of cisplatin.Therefore,reducing the renal injury caused by cisplatin is particularly important in the clinical application of cisplatin.Zhibai Dihuang Granule is a Chinese patent medicine with the function of "nourishing yin and reducing heat".At present,it is also used to treat various kidney diseases in clinic.This paper intends to study the preventive and therapeutic effect of Zhibai Dihuang Granule on cisplatin induced acute renal injury and explore its potential mechanism.This experiment preliminarily predicted the ability of Zhibai Dihuang Granule to prevent and treat cisplatin acute renal injury through network pharmacology,and analyzed the possible main active components,action targets and related mechanisms.Subsequently,the acute renal injury model of cisplatin was established at the cellular and animal levels to verify the preventive effect of Zhibai Dihuang Granule and explore the potential mechanism.The results of network pharmacology showed that Zhibai Dihuang Granule had 22 active compounds and 226 potential targets.There are 498 gene targets related to cisplatin induced acute renal injury.The intersection of the two targets to obtain 40 repeated targets may be the potential target of Zhibai Dihuang Granule in the prevention and treatment of cisplatin induced acute renal injury.The core targets include CASP3,VEGFA,IL6 and so on.After enrichment analysis,339 GO entries and 64 KEGG pathways were obtained.The main mechanisms may include DNA replication and I-κB/NF-κB pathway,external apoptosis pathway,etc.In the experimental verification section,select 25 μL The results showed that Zhibai Dihuang Granule could reduce the cytotoxicity of cisplatin to HK-2 cells and 293 T cells,and alleviate the apoptosis and the production of reactive oxygen species.Moreover,Zhibai Dihuang Granule did not affect the antitumor effect of cisplatin in A549 cells and HCT116 nuclear MCF7 cells,but had a slight synergistic antitumor effect in lung cancer A549 cells.In the rat model,10 mg/kg cisplatin caused the increase of renal index,the concentration of serum creatinine and blood urea nitrogen,and the damage of renal histopathological structure.Compared with the model group,the relevant indexes of Zhibai Dihuang Granule group were significantly relieved,indicating that Zhibai Dihuang Granule can reduce the acute renal injury induced by cisplatin.Cisplatin caused the increase of MDA level,the decrease of SOD activity and the decrease of GSH content in rat kidney.Under the action of Zhibai Dihuang Granule,it reversed the changes of oxidation indexes,indicating that Zhibai Dihuang Granule can alleviate the oxidative stress induced by cisplatin.The results of Western blot showed that the expression of cl-Caspase3 in cisplatin group was up-regulated,and the expression of cl-Caspase3 was down regulated under the action of Zhibai Dihuang Granule,indicating that Zhibai Dihuang Granule can reduce cisplatin induced apoptosis.At the same time,the results of Western blot showed that the expression of p-p65 in cisplatin group was up-regulated,and the expression of p-p65 was down regulated under the action of Zhibai Dihuang Granule,indicating that Zhibai Dihuang Granule can reduce the inflammatory response induced by cisplatin.In terms of the mechanism of action,it was verified by Western blot experiment that cisplatin could lead to the changes of protein levels related to HK-2cells and 293 T cells: the expression of anti apoptotic protein Bcl-2 was down-regulated,the expression of Pro-apoptotic proteins Bax and Bcl-x L were up-regulated,the activation of Caspase-3 was increased,and the inflammatory index: the phosphorylation level of NF-κB p65 was up-regulated,and the phosphorylation levels of makp pathway proteins JNK,p38 MAPK and Erk 1/2 increased.Zhibai Dihuang Granule can alleviate the imbalance of protein level and restore it to a relatively normal expression level.It is proved that the mechanism of Zhibai Dihuang Granule in preventing and treating cisplatin acute renal injury mainly involves MAPK pathway and NF-κB pathway and external apoptosis pathway prove that Zhibai Dihuang Granule can prevent and treat acute renal injury caused by cisplatin through a variety of cell signal pathways.This study predicted the potential of Zhibai Dihuang Granule in the prevention and treatment of cisplatin induced acute renal injury through network pharmacology,and verified and explored the pharmacodynamic basis and action mechanism through basic experiments.It is proved that Zhibai Dihuang Granule is expected to become a potential candidate drug for the prevention and treatment of cisplatin induced acute renal injury,which provides a scientific basis for the clinical application of Zhibai Dihuang Granule.