Protective Effect Of Rosmarinic Acid On Cisplatin Induced Acute Liver And Kidney Injury In Mice

Objective: In recently years,drug-induced liver and kidney injury caused by unreasonable use of drugs seriously endangers human and animal health,so it is necessary to seek natural low toxicity drugs to alleviate drug-induced liver and kidney injury.The purpose of this study was to investigate the protective effect of Rosmarinic acid on Cisplatin-induced acute liver and kidney injury in mice and its mechanism,in order to provide pharmacodynamic basis for the development of Rosmarinic acid as a drug to alleviate the drug-induced liver and kidney injury in human medicine and veterinary medicine.Methods:(1)To establish pathological model of acute liver and kidney injury in mice: Twenty male BALB/c mice were randomly divided into four groups,namely Control group and Cisplatin(10,20 and 40 mg/kg)model group,with five mice in each group.In Cisplatin model group,acute liver and kidney injury in mice was induced by one-time single dose intraperitoneal injection Cisplatin(10,20,40 mg/kg.BW),and 0.9% Na Cl solution was given to Control group at the same dose.After Seventy-two hours,blood was taken to detect the contents of ALT,AST,BUN and CRE in serum,and the liver,kidney and spleen were taken to calculate the organ coefficients and make pathological sections of the liver and kidney.(2)The protective effect of rosmarinic acid on Cisplatin induced acute liver and kidney injury in mice and its mechanism: Forty-two male BALB/c mice were randomly divided into seven groups,namely Control group,Cisplatin model group,Rosmarinic acid control group,Rosmarinic acid pretreatment groups(5,10,20 mg/kg),and Dexamethasone pretreatment group(4 mg/kg),with 6 mice in each group.The drug pretreatment groups received intraperitoneal injection of Dexamethasone(4 mg/kg.BW)or Rosmarinic acid(5,10,20 mg/kg.BW)for seven days consecutively,and mice in the Control group and Cisplatin model group were given 0.9% Na Cl solution at the same dose.On the 5th day of the experiment,Cisplatin(20 mg/kg.BW)intraperitoneal injection induced acute liver and kidney injury in mice in all groups except Control group and Rosmarinic acid control group.After seventy-two hours,blood was taken to detect the contents of ALT,AST,BUN,CRE,IL-1β,IL-6,TNF-α in serum,and the liver,kidney and spleen were taken to calculate the organ coefficients and make pathological sections of the liver and kidney,the expression levels of IL-1β,IL-6,TNF-α m RNA in liver and kidney were detected by q RT-PCR.(3)On the basis of the second part of the experiment,study on the mechanism of Rosmarinic acid on Cisplatin-induced acute liver and kidney injury in mice: the liver and kidney were taken to detect the activities of SOD,CAT,and the levels of GSH,T-AOC,MDA,NO,MPO and the m RNA expression levels of SOD,CAT,GPx,Nrf2,Keap1,HO-1,GCLC,GCLM by q RT-PCR.Results:(1)Compared with control group,each Cisplatin model group gradually appeared some clinical features such as listlessness,messy hair,slow movement,reduced urine output and even no urine;the contents of BUN,CRE,ALT,AST in serum were increased;slight vacuolar degeneration of local cells was observed in liver tissue,disorder of renal tissue structure and degeneration of proximal tubules,such as the disappearance of brush border,cytoplasmic vacuoles,and the shedding and necrosis of renal tubular epithelial cells.After comprehensive consideration,the clinical symptoms,liver and kidney function,serum biochemical indexes and histopathological results,the pathological models of acute liver and kidney injury in mice was finally established with Cisplatin of 20 mg/kg.BW.(2)Compared with control group,Cisplatin model group showed typical clinical features such as listlessness,messy hair,slow movement and Countersunk curled up;slight vacuolar degeneration of local cells was observed in liver tissue,disorder of renal tissue structure,degeneration changes of proximal tubules,disappearance of brush border,light staining of cytoplasm,pyknosis,fragmentation or dissolution of nucleus,degeneration or necrosis of renal tubular epithelial cells;the contents of BUN,CRE,ALT,AST,IL-1β,IL-6,TNF-α and the m RNA expression levels of IL-1β,IL-6 and TNF-αin liver and kidney tissue were increased.Rosmarinic acid pretreatment can reverse the clinical symptoms of mice and reduce the damage of liver and kidney tissue,and reduce the contents of BUN,CRE,ALT,AST,IL-1β,IL-6,TNF-α in serum,and inhibite the m RNA expression levels of IL-1β,IL-6 and TNF-α in liver and kidney tissue.(3)Compared with control group,in the Cisplatin model group,the levels of MPO,MDA,NO in liver and kidney tissue were increased,the activities of SOD and CAT,and the levels of GSH and T-AOC in liver and kidney tissue and the m RNA expression levels of SOD,CAT,GPx,Nrf2,Keap1,HO-1,GCLC,GCLM were decreased.Rosmarinic acid pretreatment can significantly reduce the level of MPO,MDA,NO in the liver and kidney tissue,enhance the total antioxidant capacity of liver and kidney tissues,and up-regulate the m RNA expression levels of related target genes downstream of Nrf2 signaling pathway.Conclusion: Cisplatin of 20 mg/kg.BW by one-time single dose intraperitoneal injection could successfully establish the pathological model of acute liver and kidney injury in mice.Rosmarinic acid can reduce Cisplatin-induced the acute liver and kidney injury in mice,possibly by activating Nrf2 signaling pathway to inhibit inflammatory reaction reduce oxidative stress injury and enhance antioxidant capacity of liver and kidney tissues.