Observation Of Acute Kidney Injury Induced By Different Doses Of Cisplatin In Mice Model And Detection Of Some Biomarkers

Background Acute kidney injury(AKI) is a set of clinical syndrome with clinical manifestations of decreased urine volume,azotemia,fluid balance disorders and electrolyte disorder. The common secondary causes of AKI include surgery and drugs and so on. AKI is traditionally divided into 3 categories:prerenal, intrinsic,and postrenal. Along with all kinds of drug application and environment change,AKI incidence is increasing over recent years.It is estimated that severe cases can endanger patient life.The nephrotoxic drugs have been responsible for about 20% of AKI episodes in inpatients and outpatients. Cisplatin (cis-diamminedichloroplatinum Ⅱ-CDDP) is an oncologic medication applied in most chemotherapy regimens for solid or hematologic tumors. The cisplatin nephrotoxicity is a major limiting factors in 20% of patients who have received the drug, triggering injuries in renal tubular epithelial cells. The AKI animal model was usually induced by CDDP in the experient,while the dose of CDDP was not completely unified.This experiment aims at using different doses of cisplatin to establish the AKI animal model and detect concentrations of serum creatinine and expression of heme oxygenase-1 (HO-1),regulated normal T cell expressed and secreted factors (RANTES) in the process of pathological changes.Objectives1. To observe the renal toxicity induced by different dosage of cisplatin at different time points among different groups.2. To study and explore the expressions of HO-1 and RANTES in the development of AKI disease.Methods1.60 healthy 6-8 week old male KM mouse were divided into four groups using the random number method:model groupA,model groupB,model groupC,normal control group. Model group mice were established by injecting cisplatin intraperitoneally once at a dose of 10mg/kg,15mg/kg,25mg/kg body weight respectively. According to references, it needs to be diluted with equal 0.9% sterile normal saline for the concentration of lmg/ml before the injection, so AKI animal model is set up, normal control group can be injected with same dose of sterile normal saline.2. After injected cisplatin at dayl,day3,and day5 respectively,selecting five mouse at different time points in each group randomly.The serum was collected from heart blood to measure Scr levels to evaluate renal function;fixing the kidney tissues to observe pathological changes in the kidney with hematoxylin eosin staining(HE);The levels of heme oxygenase-1(HO-1),regulated activation upon normal Tcell expressed and secreted factors(RANTES)in renal tissue were detected by enzyme-linked immunosorbent assay (ELISA).The effect of different doses on the expression of HO-1 and RANTES in renal tissue at different time were detected by immunohistochemical (IHC) staining.Results1. Compared with normal control group,a significant increase in Scr concentration happened in model group at day3 and day5,the difference were statistically significant(P< 0.05).Model groups compared with each other,elevated Scr concentration level was statistically significant(P<0.05).2. Compared with normal control group,the level of HO-1 and RANTES significantly increased in model group,the difference were statistically significant(P<0.05). Model groups compared with each other,the level of HO-1 and RANTES was statistically significant(P<0.05).3. Compared with Scr,the level of HO-1 and RANTES were increased significantly in early stage of renal injury.Conclusions1. The nephrotoxic effect of cisplatin is cumulative and dose-dependent.2. HO-1 may play a protective role in the AKI repair process by inhibiting the oxygen overload.3. RANTES is an important factor which participates in the AKI process induced by CDDP.